US20180117100A1 - Lactobacillus sp. strain having ability to inhibit proliferation of virginal pathogenic microorganisms - Google Patents
Lactobacillus sp. strain having ability to inhibit proliferation of virginal pathogenic microorganisms Download PDFInfo
- Publication number
- US20180117100A1 US20180117100A1 US15/566,352 US201615566352A US2018117100A1 US 20180117100 A1 US20180117100 A1 US 20180117100A1 US 201615566352 A US201615566352 A US 201615566352A US 2018117100 A1 US2018117100 A1 US 2018117100A1
- Authority
- US
- United States
- Prior art keywords
- strain
- lactobacillus
- sneathia
- snuv
- vaginitis
- 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.)
- Abandoned
Links
- 241000186610 Lactobacillus sp. Species 0.000 title claims abstract description 42
- 244000000010 microbial pathogen Species 0.000 title 1
- 230000035755 proliferation Effects 0.000 title 1
- 206010046914 Vaginal infection Diseases 0.000 claims abstract description 52
- 201000008100 Vaginitis Diseases 0.000 claims abstract description 52
- 241000207201 Gardnerella vaginalis Species 0.000 claims abstract description 35
- 230000001717 pathogenic effect Effects 0.000 claims abstract description 28
- 241000424747 Sneathia Species 0.000 claims abstract description 26
- 241000894006 Bacteria Species 0.000 claims abstract description 21
- 230000036541 health Effects 0.000 claims abstract description 21
- 230000000694 effects Effects 0.000 claims abstract description 20
- 235000013376 functional food Nutrition 0.000 claims abstract description 19
- 239000004480 active ingredient Substances 0.000 claims abstract description 17
- 239000008194 pharmaceutical composition Substances 0.000 claims abstract description 17
- 241000222122 Candida albicans Species 0.000 claims abstract description 15
- 229940095731 candida albicans Drugs 0.000 claims abstract description 15
- 241000701806 Human papillomavirus Species 0.000 claims abstract description 10
- 244000005700 microbiome Species 0.000 claims description 33
- 241000186840 Lactobacillus fermentum Species 0.000 claims description 20
- 241001561398 Lactobacillus jensenii Species 0.000 claims description 20
- 229940012969 lactobacillus fermentum Drugs 0.000 claims description 19
- 241000222120 Candida <Saccharomycetales> Species 0.000 claims description 17
- 241000186606 Lactobacillus gasseri Species 0.000 claims description 16
- 241000218492 Lactobacillus crispatus Species 0.000 claims description 15
- 241000711827 Gardnerella sp. Species 0.000 claims description 14
- 239000003833 bile salt Substances 0.000 claims description 12
- 239000007788 liquid Substances 0.000 claims description 11
- 241000123706 Sneathia sanguinegens Species 0.000 claims description 7
- 235000013361 beverage Nutrition 0.000 claims description 7
- 241001115883 Sneathia amnii Species 0.000 claims description 6
- 239000000344 soap Substances 0.000 claims description 6
- 241000233866 Fungi Species 0.000 claims description 3
- 241000700605 Viruses Species 0.000 claims description 3
- 230000004083 survival effect Effects 0.000 claims description 3
- 239000002537 cosmetic Substances 0.000 claims description 2
- 235000015140 cultured milk Nutrition 0.000 claims description 2
- 235000013373 food additive Nutrition 0.000 claims description 2
- 239000002778 food additive Substances 0.000 claims description 2
- 239000002453 shampoo Substances 0.000 claims description 2
- 239000007787 solid Substances 0.000 claims description 2
- 230000002401 inhibitory effect Effects 0.000 abstract description 20
- 244000052769 pathogen Species 0.000 abstract description 16
- 208000015181 infectious disease Diseases 0.000 abstract description 12
- 244000005706 microflora Species 0.000 abstract description 10
- 208000004926 Bacterial Vaginosis Diseases 0.000 abstract description 9
- 238000002955 isolation Methods 0.000 abstract description 6
- 230000002265 prevention Effects 0.000 abstract description 4
- 241000186660 Lactobacillus Species 0.000 description 65
- 229940039696 lactobacillus Drugs 0.000 description 64
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 13
- 238000002360 preparation method Methods 0.000 description 12
- 239000000203 mixture Substances 0.000 description 11
- 239000000047 product Substances 0.000 description 11
- 230000003115 biocidal effect Effects 0.000 description 10
- 239000000796 flavoring agent Substances 0.000 description 9
- 239000002609 medium Substances 0.000 description 9
- 229920001817 Agar Polymers 0.000 description 8
- 239000008272 agar Substances 0.000 description 8
- 238000004458 analytical method Methods 0.000 description 8
- 238000011156 evaluation Methods 0.000 description 8
- 238000002474 experimental method Methods 0.000 description 8
- 235000013355 food flavoring agent Nutrition 0.000 description 8
- 241000894007 species Species 0.000 description 8
- 239000003242 anti bacterial agent Substances 0.000 description 7
- 229940088710 antibiotic agent Drugs 0.000 description 7
- 230000008859 change Effects 0.000 description 7
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- 241000304886 Bacilli Species 0.000 description 6
- ULGZDMOVFRHVEP-RWJQBGPGSA-N Erythromycin Chemical compound O([C@@H]1[C@@H](C)C(=O)O[C@@H]([C@@]([C@H](O)[C@@H](C)C(=O)[C@H](C)C[C@@](C)(O)[C@H](O[C@H]2[C@@H]([C@H](C[C@@H](C)O2)N(C)C)O)[C@H]1C)(C)O)CC)[C@H]1C[C@@](C)(OC)[C@@H](O)[C@H](C)O1 ULGZDMOVFRHVEP-RWJQBGPGSA-N 0.000 description 6
- 241000192125 Firmicutes Species 0.000 description 6
- 241001468155 Lactobacillaceae Species 0.000 description 6
- 241001112724 Lactobacillales Species 0.000 description 6
- 241000699666 Mus <mouse, genus> Species 0.000 description 6
- 201000010099 disease Diseases 0.000 description 6
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 6
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 6
- 241000207202 Gardnerella Species 0.000 description 5
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 5
- 208000037009 Vaginitis bacterial Diseases 0.000 description 5
- 239000002253 acid Substances 0.000 description 5
- 210000000941 bile Anatomy 0.000 description 5
- 229940093761 bile salts Drugs 0.000 description 5
- 239000003795 chemical substances by application Substances 0.000 description 5
- -1 etc. Substances 0.000 description 5
- 235000013305 food Nutrition 0.000 description 5
- 230000005764 inhibitory process Effects 0.000 description 5
- 239000000546 pharmaceutical excipient Substances 0.000 description 5
- 210000001215 vagina Anatomy 0.000 description 5
- 238000010171 animal model Methods 0.000 description 4
- 239000004615 ingredient Substances 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 3
- 229920002472 Starch Polymers 0.000 description 3
- 239000004098 Tetracycline Substances 0.000 description 3
- 108010059993 Vancomycin Proteins 0.000 description 3
- 229960000723 ampicillin Drugs 0.000 description 3
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 150000001720 carbohydrates Chemical class 0.000 description 3
- 235000014633 carbohydrates Nutrition 0.000 description 3
- 229960002227 clindamycin Drugs 0.000 description 3
- KDLRVYVGXIQJDK-AWPVFWJPSA-N clindamycin Chemical compound CN1C[C@H](CCC)C[C@H]1C(=O)N[C@H]([C@H](C)Cl)[C@@H]1[C@H](O)[C@H](O)[C@@H](O)[C@@H](SC)O1 KDLRVYVGXIQJDK-AWPVFWJPSA-N 0.000 description 3
- 239000012228 culture supernatant Substances 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 239000003085 diluting agent Substances 0.000 description 3
- 229960003276 erythromycin Drugs 0.000 description 3
- 238000009472 formulation Methods 0.000 description 3
- 239000003966 growth inhibitor Substances 0.000 description 3
- 239000004310 lactic acid Substances 0.000 description 3
- 235000014655 lactic acid Nutrition 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 230000000813 microbial effect Effects 0.000 description 3
- 239000008107 starch Substances 0.000 description 3
- 235000019698 starch Nutrition 0.000 description 3
- 229960005322 streptomycin Drugs 0.000 description 3
- 239000000829 suppository Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 229960002180 tetracycline Drugs 0.000 description 3
- 229930101283 tetracycline Natural products 0.000 description 3
- 235000019364 tetracycline Nutrition 0.000 description 3
- 229960003165 vancomycin Drugs 0.000 description 3
- 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 3
- MYPYJXKWCTUITO-LYRMYLQWSA-O vancomycin(1+) 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)[NH2+]C)[C@H]1C[C@](C)([NH3+])[C@H](O)[C@H](C)O1 MYPYJXKWCTUITO-LYRMYLQWSA-O 0.000 description 3
- 108020004465 16S ribosomal RNA Proteins 0.000 description 2
- 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 2
- 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 2
- 108020004414 DNA Proteins 0.000 description 2
- 238000007400 DNA extraction Methods 0.000 description 2
- 239000004386 Erythritol Substances 0.000 description 2
- UNXHWFMMPAWVPI-UHFFFAOYSA-N Erythritol Natural products OCC(O)C(O)CO UNXHWFMMPAWVPI-UHFFFAOYSA-N 0.000 description 2
- 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 2
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- 208000009608 Papillomavirus Infections Diseases 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- 229930003448 Vitamin K Natural products 0.000 description 2
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 239000002671 adjuvant Substances 0.000 description 2
- 235000010443 alginic acid Nutrition 0.000 description 2
- 229920000615 alginic acid Polymers 0.000 description 2
- 239000003125 aqueous solvent Substances 0.000 description 2
- 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 2
- 239000011230 binding agent Substances 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 239000000969 carrier Substances 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 229960005091 chloramphenicol Drugs 0.000 description 2
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 2
- 238000003501 co-culture Methods 0.000 description 2
- 239000003086 colorant Substances 0.000 description 2
- 239000006071 cream Substances 0.000 description 2
- 238000009792 diffusion process Methods 0.000 description 2
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 239000000839 emulsion Substances 0.000 description 2
- 235000019414 erythritol Nutrition 0.000 description 2
- UNXHWFMMPAWVPI-ZXZARUISSA-N erythritol Chemical compound OC[C@H](O)[C@H](O)CO UNXHWFMMPAWVPI-ZXZARUISSA-N 0.000 description 2
- 229940009714 erythritol Drugs 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 235000015203 fruit juice Nutrition 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 208000021145 human papilloma virus infection Diseases 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 238000003780 insertion Methods 0.000 description 2
- 230000037431 insertion Effects 0.000 description 2
- 239000008101 lactose Substances 0.000 description 2
- 239000000314 lubricant Substances 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 2
- 229960000282 metronidazole Drugs 0.000 description 2
- VAOCPAMSLUNLGC-UHFFFAOYSA-N metronidazole Chemical compound CC1=NC=C([N+]([O-])=O)N1CCO VAOCPAMSLUNLGC-UHFFFAOYSA-N 0.000 description 2
- 239000006872 mrs medium Substances 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- 239000002674 ointment Substances 0.000 description 2
- 239000002953 phosphate buffered saline Substances 0.000 description 2
- SHUZOJHMOBOZST-UHFFFAOYSA-N phylloquinone Natural products CC(C)CCCCC(C)CCC(C)CCCC(=CCC1=C(C)C(=O)c2ccccc2C1=O)C SHUZOJHMOBOZST-UHFFFAOYSA-N 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 239000003755 preservative agent Substances 0.000 description 2
- 238000000513 principal component analysis Methods 0.000 description 2
- 239000006041 probiotic Substances 0.000 description 2
- 230000000529 probiotic effect Effects 0.000 description 2
- 235000018291 probiotics Nutrition 0.000 description 2
- HELXLJCILKEWJH-NCGAPWICSA-N rebaudioside A Chemical compound O([C@H]1[C@H](O)[C@@H](CO)O[C@H]([C@@H]1O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)O[C@]12C(=C)C[C@@]3(C1)CC[C@@H]1[C@@](C)(CCC[C@]1([C@@H]3CC2)C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O HELXLJCILKEWJH-NCGAPWICSA-N 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 239000000600 sorbitol Substances 0.000 description 2
- 235000010356 sorbitol Nutrition 0.000 description 2
- 239000003381 stabilizer Substances 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 239000000375 suspending agent Substances 0.000 description 2
- OFVLGDICTFRJMM-WESIUVDSSA-N tetracycline Chemical compound C1=CC=C2[C@](O)(C)[C@H]3C[C@H]4[C@H](N(C)C)C(O)=C(C(N)=O)C(=O)[C@@]4(O)C(O)=C3C(=O)C2=C1O OFVLGDICTFRJMM-WESIUVDSSA-N 0.000 description 2
- 230000001515 vagal effect Effects 0.000 description 2
- 235000019168 vitamin K Nutrition 0.000 description 2
- 239000011712 vitamin K Substances 0.000 description 2
- 150000003721 vitamin K derivatives Chemical class 0.000 description 2
- 229940046010 vitamin k Drugs 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 235000010447 xylitol Nutrition 0.000 description 2
- 239000000811 xylitol Substances 0.000 description 2
- 229960002675 xylitol Drugs 0.000 description 2
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 2
- HSINOMROUCMIEA-FGVHQWLLSA-N (2s,4r)-4-[(3r,5s,6r,7r,8s,9s,10s,13r,14s,17r)-6-ethyl-3,7-dihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1h-cyclopenta[a]phenanthren-17-yl]-2-methylpentanoic acid Chemical compound C([C@@]12C)C[C@@H](O)C[C@H]1[C@@H](CC)[C@@H](O)[C@@H]1[C@@H]2CC[C@]2(C)[C@@H]([C@H](C)C[C@H](C)C(O)=O)CC[C@H]21 HSINOMROUCMIEA-FGVHQWLLSA-N 0.000 description 1
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- UAIUNKRWKOVEES-UHFFFAOYSA-N 3,3',5,5'-tetramethylbenzidine Chemical compound CC1=C(N)C(C)=CC(C=2C=C(C)C(N)=C(C)C=2)=C1 UAIUNKRWKOVEES-UHFFFAOYSA-N 0.000 description 1
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 description 1
- 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
- 241001153886 Ami Species 0.000 description 1
- 108010011485 Aspartame Proteins 0.000 description 1
- 208000031504 Asymptomatic Infections Diseases 0.000 description 1
- 241001633064 Atopobium vaginae Species 0.000 description 1
- 238000011725 BALB/c mouse Methods 0.000 description 1
- 108020000946 Bacterial DNA Proteins 0.000 description 1
- 241000321538 Candidia Species 0.000 description 1
- 229920000858 Cyclodextrin Polymers 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 239000004097 EU approved flavor enhancer Substances 0.000 description 1
- 241001394620 Eggerthella sp. Species 0.000 description 1
- LVGKNOAMLMIIKO-UHFFFAOYSA-N Elaidinsaeure-aethylester Natural products CCCCCCCCC=CCCCCCCCC(=O)OCC LVGKNOAMLMIIKO-UHFFFAOYSA-N 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 239000001512 FEMA 4601 Substances 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- 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 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 229930182566 Gentamicin Natural products 0.000 description 1
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 description 1
- 239000004378 Glycyrrhizin Substances 0.000 description 1
- 229920000084 Gum arabic Polymers 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- JVTAAEKCZFNVCJ-UHFFFAOYSA-M Lactate Chemical compound CC(O)C([O-])=O JVTAAEKCZFNVCJ-UHFFFAOYSA-M 0.000 description 1
- 241000015236 Lactobacillus fornicalis Species 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
- 241000124008 Mammalia Species 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 241001576959 Megasphaera sp. Species 0.000 description 1
- 241000736262 Microbiota Species 0.000 description 1
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 1
- 241001464887 Parvimonas micra Species 0.000 description 1
- 206010034133 Pathogen resistance Diseases 0.000 description 1
- 229920002230 Pectic acid Polymers 0.000 description 1
- 102000003992 Peroxidases Human genes 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 208000005107 Premature Birth Diseases 0.000 description 1
- 241001135215 Prevotella bivia Species 0.000 description 1
- HELXLJCILKEWJH-SEAGSNCFSA-N Rebaudioside A Natural products O=C(O[C@H]1[C@@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1)[C@@]1(C)[C@@H]2[C@](C)([C@H]3[C@@]4(CC(=C)[C@@](O[C@H]5[C@H](O[C@H]6[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O6)[C@@H](O[C@H]6[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O6)[C@H](O)[C@@H](CO)O5)(C4)CC3)CC2)CCC1 HELXLJCILKEWJH-SEAGSNCFSA-N 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- 241000191940 Staphylococcus Species 0.000 description 1
- 244000228451 Stevia rebaudiana Species 0.000 description 1
- 241000224526 Trichomonas Species 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000000205 acacia gum Substances 0.000 description 1
- 235000010489 acacia gum Nutrition 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 229940072056 alginate Drugs 0.000 description 1
- 239000000783 alginic acid Substances 0.000 description 1
- 229960001126 alginic acid Drugs 0.000 description 1
- 150000004781 alginic acids Chemical class 0.000 description 1
- 230000000843 anti-fungal effect Effects 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000000605 aspartame Substances 0.000 description 1
- 235000010357 aspartame Nutrition 0.000 description 1
- IAOZJIPTCAWIRG-QWRGUYRKSA-N aspartame Chemical compound OC(=O)C[C@H](N)C(=O)N[C@H](C(=O)OC)CC1=CC=CC=C1 IAOZJIPTCAWIRG-QWRGUYRKSA-N 0.000 description 1
- 229960003438 aspartame Drugs 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
- 239000003613 bile acid Substances 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000006172 buffering agent Substances 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 239000000378 calcium silicate Substances 0.000 description 1
- 229910052918 calcium silicate Inorganic materials 0.000 description 1
- 235000012241 calcium silicate Nutrition 0.000 description 1
- OYACROKNLOSFPA-UHFFFAOYSA-N calcium;dioxido(oxo)silane Chemical compound [Ca+2].[O-][Si]([O-])=O OYACROKNLOSFPA-UHFFFAOYSA-N 0.000 description 1
- 235000014171 carbonated beverage Nutrition 0.000 description 1
- 238000010000 carbonizing Methods 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 235000010980 cellulose Nutrition 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 210000003756 cervix mucus Anatomy 0.000 description 1
- 235000013351 cheese Nutrition 0.000 description 1
- 239000013043 chemical agent Substances 0.000 description 1
- 235000019219 chocolate Nutrition 0.000 description 1
- 239000007330 chocolate agar Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 239000007382 columbia agar Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 150000002016 disaccharides Chemical class 0.000 description 1
- 239000002270 dispersing agent Substances 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- HELXLJCILKEWJH-UHFFFAOYSA-N entered according to Sigma 01432 Natural products C1CC2C3(C)CCCC(C)(C(=O)OC4C(C(O)C(O)C(CO)O4)O)C3CCC2(C2)CC(=C)C21OC(C1OC2C(C(O)C(O)C(CO)O2)O)OC(CO)C(O)C1OC1OC(CO)C(O)C(O)C1O HELXLJCILKEWJH-UHFFFAOYSA-N 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- LVGKNOAMLMIIKO-QXMHVHEDSA-N ethyl oleate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC LVGKNOAMLMIIKO-QXMHVHEDSA-N 0.000 description 1
- 229940093471 ethyl oleate Drugs 0.000 description 1
- 210000001752 female genitalia Anatomy 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 239000006260 foam Substances 0.000 description 1
- 235000019264 food flavour enhancer Nutrition 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 244000053095 fungal pathogen Species 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- LPLVUJXQOOQHMX-UHFFFAOYSA-N glycyrrhetinic acid glycoside Natural products C1CC(C2C(C3(CCC4(C)CCC(C)(CC4C3=CC2=O)C(O)=O)C)(C)CC2)(C)C2C(C)(C)C1OC1OC(C(O)=O)C(O)C(O)C1OC1OC(C(O)=O)C(O)C(O)C1O LPLVUJXQOOQHMX-UHFFFAOYSA-N 0.000 description 1
- 229960004949 glycyrrhizic acid Drugs 0.000 description 1
- UYRUBYNTXSDKQT-UHFFFAOYSA-N glycyrrhizic acid Natural products CC1(C)C(CCC2(C)C1CCC3(C)C2C(=O)C=C4C5CC(C)(CCC5(C)CCC34C)C(=O)O)OC6OC(C(O)C(O)C6OC7OC(O)C(O)C(O)C7C(=O)O)C(=O)O UYRUBYNTXSDKQT-UHFFFAOYSA-N 0.000 description 1
- 235000019410 glycyrrhizin Nutrition 0.000 description 1
- LPLVUJXQOOQHMX-QWBHMCJMSA-N glycyrrhizinic acid Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@H](O[C@@H]1O[C@@H]1C([C@H]2[C@]([C@@H]3[C@@]([C@@]4(CC[C@@]5(C)CC[C@@](C)(C[C@H]5C4=CC3=O)C(O)=O)C)(C)CC2)(C)CC1)(C)C)C(O)=O)[C@@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O LPLVUJXQOOQHMX-QWBHMCJMSA-N 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 230000009422 growth inhibiting effect Effects 0.000 description 1
- 229940025294 hemin Drugs 0.000 description 1
- BTIJJDXEELBZFS-QDUVMHSLSA-K hemin Chemical compound CC1=C(CCC(O)=O)C(C=C2C(CCC(O)=O)=C(C)\C(N2[Fe](Cl)N23)=C\4)=N\C1=C/C2=C(C)C(C=C)=C3\C=C/1C(C)=C(C=C)C/4=N\1 BTIJJDXEELBZFS-QDUVMHSLSA-K 0.000 description 1
- 230000003054 hormonal effect Effects 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 208000027866 inflammatory disease Diseases 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 239000007951 isotonicity adjuster Substances 0.000 description 1
- 235000015110 jellies Nutrition 0.000 description 1
- 239000008274 jelly Substances 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 239000000845 maltitol Substances 0.000 description 1
- 235000010449 maltitol Nutrition 0.000 description 1
- VQHSOMBJVWLPSR-WUJBLJFYSA-N maltitol Chemical compound OC[C@H](O)[C@@H](O)[C@@H]([C@H](O)CO)O[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O VQHSOMBJVWLPSR-WUJBLJFYSA-N 0.000 description 1
- 229940035436 maltitol Drugs 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 239000013028 medium composition Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000009245 menopause Effects 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 229920000609 methyl cellulose Polymers 0.000 description 1
- 239000001923 methylcellulose Substances 0.000 description 1
- 235000010981 methylcellulose Nutrition 0.000 description 1
- LXCFILQKKLGQFO-UHFFFAOYSA-N methylparaben Chemical compound COC(=O)C1=CC=C(O)C=C1 LXCFILQKKLGQFO-UHFFFAOYSA-N 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 1
- 239000008108 microcrystalline cellulose Substances 0.000 description 1
- 229940016286 microcrystalline cellulose Drugs 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 239000002480 mineral oil Substances 0.000 description 1
- 235000010446 mineral oil Nutrition 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000000897 modulatory effect Effects 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 238000007481 next generation sequencing Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000004006 olive oil Substances 0.000 description 1
- 235000008390 olive oil Nutrition 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 230000027758 ovulation cycle Effects 0.000 description 1
- 238000007911 parenteral administration Methods 0.000 description 1
- LCLHHZYHLXDRQG-ZNKJPWOQSA-N pectic acid Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)O[C@H](C(O)=O)[C@@H]1OC1[C@H](O)[C@@H](O)[C@@H](OC2[C@@H]([C@@H](O)[C@@H](O)[C@H](O2)C(O)=O)O)[C@@H](C(O)=O)O1 LCLHHZYHLXDRQG-ZNKJPWOQSA-N 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 239000010318 polygalacturonic acid Substances 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 150000004804 polysaccharides Chemical class 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- QELSKZZBTMNZEB-UHFFFAOYSA-N propylparaben Chemical compound CCCOC(=O)C1=CC=C(O)C=C1 QELSKZZBTMNZEB-UHFFFAOYSA-N 0.000 description 1
- 229960003415 propylparaben Drugs 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 235000019203 rebaudioside A Nutrition 0.000 description 1
- 235000019204 saccharin Nutrition 0.000 description 1
- CVHZOJJKTDOEJC-UHFFFAOYSA-N saccharin Chemical compound C1=CC=C2C(=O)NS(=O)(=O)C2=C1 CVHZOJJKTDOEJC-UHFFFAOYSA-N 0.000 description 1
- 229940081974 saccharin Drugs 0.000 description 1
- 239000000901 saccharin and its Na,K and Ca salt Substances 0.000 description 1
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical compound O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 229960002920 sorbitol Drugs 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000005846 sugar alcohols Chemical class 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 235000012222 talc Nutrition 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 150000003522 tetracyclines Chemical class 0.000 description 1
- 239000000892 thaumatin Substances 0.000 description 1
- 235000010436 thaumatin Nutrition 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 238000007473 univariate analysis Methods 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 208000010484 vulvovaginitis Diseases 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
- 239000007218 ym medium Substances 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
-
- 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
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
- A23C9/1234—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt characterised by using a Lactobacillus sp. other than Lactobacillus Bulgaricus, including Bificlobacterium sp.
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/52—Adding ingredients
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/40—Complete food formulations for specific consumer groups or specific purposes, e.g. infant formula
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
- A61P15/02—Drugs for genital or sexual disorders; Contraceptives for disorders of the vagina
-
- 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
- A61P31/10—Antimycotics
-
- 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
- A61P31/20—Antivirals for DNA viruses
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D9/00—Compositions of detergents based essentially on soap
- C11D9/04—Compositions of detergents based essentially on soap containing compounding ingredients other than soaps
-
- 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
-
- C12R1/225—
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/143—Fermentum
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/145—Gasseri
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/149—Jensenii
-
- A23Y2220/00—
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
Definitions
- the present invention activity relates to a Lactobacillus sp. strain having an ability to inhibit the growth of pathogenic vaginal microorganisms, a growth inhibitor of pathogenic vaginal microorganisms including the same as an active ingredient, and a product for improving, preventing or treating vaginitis including the inhibitor. More specifically, the present invention relates to a novel Lactobacillus sp. strain having an inhibitory effect on the growth of pathogenic vaginal microorganisms wherein the pathogenic vaginal microorganisms include fungi, bacteria and viruses, and specific examples thereof include Candidia spp. strains, Gardnerella spp. strains, or Sneathia spp. strains, or the like, and a pharmaceutical composition, a health functional food or a cleansing product including the same as an active ingredient.
- bacterial vaginosis BV
- candida vaginitis CV
- trichomonas vulvovaginitis a type and causes of vaginitis caused by infection
- BV bacterial vaginosis
- CV candida vaginitis
- trichomonas vulvovaginitis a type and causes of vaginitis caused by infection
- the bacterial vaginosis has been recognized as the most common vaginitis, and it has been reported that Atopobium vaginae, Megasphaera sp., Gardnerella vaginalis, Eggerthella sp., Clostridium -like sp., Prevotella bivia, Peptostreptococcus micros , or the like have been found in the vaginal secretions isolated from patients with bacterial vaginosis.
- Gardnerella vaginalis is known as the major causative bacteria of bacterial vaginosis.
- vaginal microflora lactate-producing Lactobacillus sp.
- pathogens causing female diseases including vaginitis by performing functions, such as maintaining vaginal acidity, producing hydrogen peroxide and activating mucosal immune system, and the like.
- Typical causative bacteria of vaginitis reported so far are Gardnerella vaginalis and Candida albicans , but recently, it has been reported that Sneathia spp. strains are closely associated with the infection of human papillomavirus and preterm birth, that they may also act as vaginitis pathogens in the disturbed vaginal microflora. Accordingly, there is a need to find Lactobacillus lactic acid bacteria having an inhibitory effect on the growth of Sneathia spp. pathogens, in addition to previously known pathogens for vaginitis.
- 190 strains of Lactobacillus spp. isolated from healthy women were screened for the inhibitory activity of pathogenic vaginal microorganisms.
- Probiotic Lactobacillus strains having novel activity have been found through the detection of Lactobacillus strains having an inhibitory effect against Sneathia spp. strains, which have been reported as microbiological markers associated with human papillomavirus infection and vaginitis-inducing causative bacteria, and a comprehensive inhibitory effect on the occurrence of vaginitis has been confirmed.
- It is still another object of the present invention to provide a method for preventing and/or treating diseases associated with the growth of pathogenic vaginal microorganisms which comprises administering the Lactobacillus sp. strain to a patient having diseases associated with the growth of pathogenic vaginal microorganisms.
- the present invention provides a novel Lactobacillus sp. strain having an inhibitory effect on the growth of pathogenic vaginal microorganisms, and a pharmaceutical composition, a health functional food or a cleansing product comprising the same as an active ingredient.
- One embodiment of the present invention relates to a Lactobacillus sp. strain which is characterized by having an inhibitory activity on the growth of pathogenic vaginal microorganisms.
- the Lactobacillus sp. strain is isolated from the vaginal microflora of healthy women, and for example, it may be at least one Lactobacillus sp. strain selected from the group consisting of Lactobacillus crispatus SNUV 220 , Lactobacillus fermentum SNUV 175 , Lactobacillus jensenii SNUV 360, and Lactobacillus gasseri SNUV 281.
- the pathogenic vaginal microorganism may be at least one selected from the group consisting of fungi, bacteria and viruses.
- it may be at least one selected from the group consisting of a Candida sp. strain being a causative bacteria of Candidal vaginitis, Sneathia spp. strains associated with the occurrence of bacterial vaginitis, a Gardnerella sp. strain, a vaginitis pathogen, and human papillomavirus associated with human papillomavirus infection.
- the Lactobacillus spp. strains may exhibit growth inhibitory effects against each of these strains or simultaneously and thus can be used for the prevention and/or treatment of female vaginitis.
- the Candida sp. strain may be Candida albicans , but is not limited thereto.
- the Gardnerella sp. strain may be Gardnerella vaginalis , but is not limited thereto.
- the Sneathia spp. strain may be Sneathia amnii and Sneathia sanguinegens , but is not limited thereto.
- Lactobacillus sp. strain may function to actively produce hydrogen peroxide, and may also have acid resistance and bile resistance, but is not limited thereto.
- the hydrogen peroxide activity of the Lactobacillus sp. strain can be qualitatively discriminated for the strains that induces dark green color change, when cultured in TMB medium in which 3,3′, 5,5′-tetramethylbenzidine (250 mg/L), hemin (5 mg/L) and vitamin K (0.5 ug/L) are added to MRS agar medium for Lactobacillus culture.
- the acid resistance of the strains is determined based on the change in the growth rate when cultured in an acidic broth having a pH 2 and a pH 3 for 24 hours, and the bile resistance is determined based on the change in the growth rate when cultured for 24 hours in a broth supplemented with bile salt with a concentration ranging from 0.1% to 4% (w/v).
- the Lactobacillus sp. strain may exhibit a survival rate of 50% or higher at a concentration of 0.1% (w/v) bile salt, but is not limited thereto.
- Another embodiment of the present invention relates to a growth inhibitor of pathogenic vaginal microorganisms containing Lactobacillus sp. strain as an active ingredient, and a pharmaceutical composition for treating and/or preventing vaginitis, a food composition or cleansing product for improving, preventing or treating vaginitis including the inhibitor.
- the Lactobacillus sp. strain may be Lactobacillus crispatus, Lactobacillus fermentum, Lactobacillus jensenii , and Lactobacillus gasseri.
- the strain may be at least one Lactobacillus sp. strain selected from the group consisting of Lactobacillus crispatus SNUV 220 deposited under accession No. KCTC18374P, Lactobacillus fermentum SNUV 175 deposited under accession No. KCTC18371P, Lactobacillus jensenii SNUV 360 deposited under accession No. KCTC18372P, Lactobacillus gasseri SNUV 281 deposited under accession No. KCTC18375P
- the growth inhibitor of vaginitis pathogens containing the Lactobacillus sp. strain according to the present invention as an active ingredient exhibits antiviral activity, antifungal activity against fungal pathogens, and antimicrobial activity against bacteria, and the vaginitis pathogens may be caused by at least one strain selected from the group consisting of a Candida sp. strain, a Gardnerella sp. strain, and a Sneathia spp. strain.
- the Candida sp. strain may be Candida albicans , but is not limited thereto.
- the Gardnerella sp. strain may be Gardnerella vaginalis , but is not limited thereto.
- the Sneathia spp. strain may be Sneathia amnii and Sneathia sanguinegens , but is not limited thereto
- the pharmaceutical composition according to the present invention can be administered to mammals including human via various routes.
- the mode of administration may be any mode commonly used in the art.
- it may be administered by oral, transdermal, intravenous, intramuscular, subcutaneous routes or the like, and preferably, it may be orally administered.
- the pharmaceutical composition of the present invention may be used after being formulated into an oral preparation, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, etc., and a parental preparation, such as epidermal formulations, suppositories, or sterile injection solutions, in accordance with a conventional method
- composition of the present invention may further contain pharmaceutically suitable and physiologically acceptable adjuvants such as carriers, excipients and diluents, etc.
- Examples of carriers, excipients and diluents that can be included in the pharmaceutical composition of the present invention may include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil.
- a diluting agent or an excipient such as commonly-used fillers, weighting agents, binding agents, wetting agents, disintegrating agents, and surfactants can be used.
- topical preparations such as liquid preparation, gel preparation, cleansing composition, tablet for vagal insertion, suppository, cream, ointment, dressing solution, spray, other coating agents, etc.
- liquid preparations such as solution-type, suspension-type, emulsion-type preparation, etc.
- external skin formulations such as sterile aqueous solution, non-aqueous solvent, suspension, emulsion, freeze-drying preparation, suppository, cream, ointment, jelly, foam, cleansing agent or vaginal insert, preferably, liquid preparation, gel preparation, cleansing composition, tablet for vagal insertion, etc.
- the formulation can be prepared, for example, by adding a dissolution adjuvant, an emulsifying agent, a buffering agent for pH control, etc., to sterilized water.
- the non-aqueous solvents and suspending agents may include vegetable oils such as propylene glycol, polyethylene glycol and olive oil, and an injectable ester such as ethyl oleate and the like.
- the pharmaceutical composition of the present invention may be administered alone, but considering the mode of administration and the standard pharmaceutical practice, it can be generally administered by mixing with the selected pharmaceutical carrier.
- the composition containing the Lactobacillus sp. strain of the present invention may be orally, intrabuccally, or sublingually administered in a tablet form containing starch or lactose, in a capsule form containing only the active ingredient according to the present invention or containing an excipient in addition to the active ingredient, or in an elixir or suspension form containing a chemical agent for flavor or color.
- the dose of the pharmaceutical composition containing the Lactobacillus sp. strain of the present invention may vary depending on the patient's age, weight, sex, dosage form, health condition and severity of disease. In addition, it can be administered in divided doses once to several times a day at fixed time intervals according to the decision of a doctor or pharmacist.
- the daily dose may be 0.1 to 500 mg/kg, preferably 0.5 to 300 mg/kg, based on the content of the active ingredient.
- the above doses are exemplified as an average case, and its dose may increase or decrease depending on individual differences. If the daily dose of the composition containing the mixture extract of the present invention is less than the above-mentioned dose, a significant effect cannot be obtained. If the daily dose exceeds the above-mentioned range, not only it is non-economical, but also it may cause undesirable side effects as the dose deviates from the above range.
- Still another embodiment of the present invention provides a health functional food for improving, treating and/or preventing vaginitis containing the Lactobacillus sp. strain as an active ingredient.
- the health functional food may be various beverages, fermented milk, food additives, and the like.
- the Lactobacillus sp. strain is as described above.
- the content of the Lactobacillus sp. strain as an effective ingredient contained in the health functional food is not particularly limited, but may appropriately vary depending on the form of food, desired use or the like. For example, it may be added in an amount of 0.01 to 15% by weight based on the total weight of the food, and the health beverage composition may be added at a ratio of 0.02 to 10 g, preferably 0.3 to 1 g, based on 100 ml.
- the liquid ingredient there is no particular limitation on the liquid ingredient, except that the Lactobacillus strain is contained as an essential ingredient at the indicated ratio, and various flavoring agents or natural carbohydrates may be contained as additional ingredients as in common beverages.
- natural carbohydrates examples include common sugars including monosaccharides, such as glucose, fructose, etc., disaccharides, such as maltose, sucrose, etc., and polysaccharides, such as dextrin, cyclodextrin, etc., as well as sugar alcohols, such as xylitol, sorbitol, erythritol, etc.
- sugar alcohols such as xylitol, sorbitol, erythritol, etc.
- natural flavoring agents thaumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) may be favorably used.
- the ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g, per 100 ml of the composition of the present invention.
- the health functional food of the present invention may contain various nutrients, vitamins, minerals (electrolyte), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and flavor enhancers (cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickening agents, pH controlling agents, stabilizing agents, preservatives, glycerin, alcohol, carbonizing agents as used in carbonated beverages and the like.
- flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and flavor enhancers (cheese, chocolate, etc.
- pectic acid and salts thereof such as synthetic flavoring agents and natural flavoring agents, coloring agents and flavor enhancers (cheese, chocolate, etc.
- pectic acid and salts thereof such as synthetic flavoring agents and natural flavoring agents, coloring agents and flavor enhancers (cheese, chocolate, etc.
- pectic acid and salts thereof such as synthetic flavoring agents and natural flavoring agents
- the health functional food of the present invention may contain fruits, as used in preparing natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. Although the proportion of these additives is not of great importance, it is generally selected from a range of 0 to about 20 parts by weight per 100 parts by weight of the health functional food of the present invention.
- Still another embodiment of the present invention provides a cleansing product for improving, treating and/or preventing vaginitis containing the Lactobacillus sp. strain as an active ingredient.
- the cleansing product may be a solid cosmetic soap, a hand cleaner, a liquid shampoo, a liquid soap, a liquid conditioner, a body cleaner, a creamy soap, or the like.
- the Lactobacillus sp. strain is as described above.
- the cleansing product may further include a carrier for formulating a preparation.
- a carrier for formulating a preparation examples include a binding agent, a lubricant, an agent for suspension, a solubilizing agent, a buffer, a preservative, a lubricant, an isotonic agent, an excipient, a stabilizer, a dispersant, a suspending agent, a coloring agent, a flavoring agent, etc.
- the present invention relates to a novel Lactobacillus sp. strain having an ability to inhibit the growth of pathogenic vaginal microorganisms, and a pharmaceutical composition, a health functional food, and a cleansing product containing the same as an active ingredient. Therefore, the present invention exhibits an activity to inhibit the growth of Sneathia spp. pathogens associated with the human papillomavirus (HPV) infection and the occurrence of bacterial vaginitis, and an inhibitory effect on the growth of Gardnerella vaginalis which is a vaginitis pathogen, and Candida albicans which is a causative bacteria of Candidal vaginitis, and thus can be used for the improvement, prevention and/or treatment of female vaginitis.
- HPV human papillomavirus
- FIG. 1 is a graph showing the measurement results of bile resistance by confirming the growth rate after culturing for 24 hours in a medium containing 0.1% to 4% (w/v) of bile salt according to one embodiment of the present invention.
- FIG. 2 is a representative graph showing the measurement results of antibiotic resistance of Lactobacillus fermentum SNUV 175 and Lactobacillus jensenii SNUV 360 according to one embodiment of the present invention against nine antibiotics.
- FIG. 3 is a graph showing the relative abundance of Gardnerella vaginalis for each treatment group, which is the result of microbiome analysis after vaginal administration of four types of Lactobacillus strains according to one embodiment of the present invention into a mouse animal model infected with Gardnerella vaginalis.
- FIG. 4 is a principal component analysis (PCoA) graph showing the change in the community structure of vaginal microflora after 2 days of administration, which are the results of microbiome analysis after vaginal administration of four types of Lactobacillus strains according to one embodiment of the present invention into a mouse animal model infected with Gardnerella vaginalis.
- PCoA principal component analysis
- FIG. 5 is a graph showing the results of univariate analysis of microorganism groups with a significant change for each group, which are the results of microbiome analysis after vaginal administration of four types of Lactobacillus strains according to one embodiment of the present invention into a mouse animal model infected with Gardnerella vaginalis.
- vaginal microflora About 190 different Lactobacillus strains were isolated from the vaginal microflora of healthy women. Specifically, the samples for the isolation of vaginal microflora were obtained from nine subjects, including three pairs of identical twins and their mothers who participated in the Korean Twin-Family Cohort Study, and were supplied from Samsung Hospital in the form of mid-vaginal swab samples (IRB No. 144-2011-07-11).
- the swab samples were transferred to the present laboratory in the form of being stored in a refrigerator within 4 hours after collection in a modified Liquid Amies solution and immediately used for the isolation of strains.
- the samples were sequentially diluted from 10 ⁇ 1 times to 10 ⁇ 8 times and spread on three different media of Chocolate agar, Rogosa agar, and Columbia agar, and cultured for 48 hours under anaerobic conditions.
- PCR reaction was carried out using UnivFwd (5′-AGA GTT TGA TCM TGG CTC AG-3′) primer and UnivRev (5′-GGY TAC CTT GTT ACG ACT T-3′) primer for 16S ribosomal RNA typing.
- the PCR products were purified using QIAquick PCR purification kit and subjected to sequence analysis using ABI3711 automatic sequencer.
- the results were found to be Lactobacillus crispatus, Lactobacillus fermentum, Lactobacillus jensenii , and Lactobacillus gasseri , respectively. Accordingly, they were assigned SNUV 220, SNUV 175, SNUV 360, and SNUV 281, respectively, and deposited with the Korean Collection for Type Cultures (KCTC) located in Yuseong-gu, Daejeon, South Korea and were assigned accession numbers KCTC18374P (SNUV 220, deposited on April 9), KCTC18371P (SNUV 175, deposited on April 7), KCTC18372P (SNUV 360, deposited on April 7) and KCTC18375P (SNUV 281, deposited on April 9).
- KCTC18374P accession numbers
- KCTC18371P SNUV 175, deposited on April 7
- KCTC18372P SNUV 360, deposited on April 7
- KCTC18375P SNUV 281, deposited on April 9
- glycerol (16% v/v) was added to the culture medium that reached the exponential phase, and stored at ⁇ 80° C. as a stock.
- 1% of each strain was inoculated in an anaerobic MRS medium and cultured at 37° C. for 24 hours. Then, the microbial cells were removed by centrifugation at 13000 ⁇ g for 10 minutes, and the supernatant was passed through a membrane filter with a pore size of 0.22 um and then stored at ⁇ 80° C. until they were used in the experiment.
- Lactobacillus strains isolated in Example 1 was used for the assay by culturing at 37° C. for 20 hours using MRS broth (Difco, USA).
- MRS broth Difco, USA.
- the strains of Sneathia spp. and Gardnerella vaginalis were inoculated into NYCIII broth and then anaerobically cultured at 37° C. for 48 hours or 24 hours, respectively, and used in the experiment.
- Sneathia spp. strains used in the experiment two kinds of strains of Sneathia spp., which was isolated in Virginia Commonwealth University School of Medicine and has been reported in the paper, and Sneathia sanguinegens that the present inventors isolated from Korean women were used.
- KCTC 5096 strain furnished from the Korean Collection for Type Cultures (KCTC) was used.
- the inhibitory activity was expressed as the diameter (mm) of the transparent disk in which the growth of the strains was inhibited from the center of the disk.
- the disk inhibition assay was performed against the strains of Sneathia spp. and Gardnerella vaginalis with culture supernatant of isolated Lactobacillus strains, and the representative result are shown in Table 4 below.
- Lactobacillus crispatus SNUV 220 Lactobacillus fermentum SNUV 175 , Lactobacillus jensenii SNUV 360, and Lactobacillus gasseri SNUV 281 were selected.
- Lactobacillus jensenii SNUV 360 strain exhibited a most potent inhibitory activity against both strains of Sneathia spp. and Gardnerella sp.
- Candida albicans strain which is a causative bacteria of Candidal vaginitis of the four kinds of Lactobacillus strains screened in Example 2 above
- Candida albicans ATCC 44858 strain American Type Culture Collection
- MYA4788 strain American Type Culture Collection
- Candida albicans strains were evaluated using a 96-well diffusion test.
- 50 ul of Candida albicans ATCC44858 (or Candida albicans M4788) culture media diluted by adding 100 ul of YM medium and 100 ul of Lactobacillus culture supernatant was added to a well of 96-well plate, and then cultured at 37° C. for 24 hours. Thereafter, the growth of inhibited Candida strains was estimated by measurement of absorbance at 600 nm, and the results are shown in Table 5.
- TMB agar medium was prepared as shown in Table 6 below (medium composition per 1 L).
- Lactobacillus crispatus SNUV 220 Lactobacillus fermentum SNUV 175 , Lactobacillus jensenii SNUV 360, and Lactobacillus gasseri SNUV 281, which are the four types of Lactobacillus strains screened in Example 2, was cultured in MRS broth at 37° C. for 20 hours, inoculated onto the TMB agar plate, and then anaerobically cultured at 37° C. for 2 days. After the culture, the plate was exposed to the air for 30 minutes and evaluated via a qualitative experiment in which the color of the colonies turns to blue. The extent to which the color of colonies turned to blue was observed with the naked eye, and the results are shown in Table 7 below.
- the hydrogen peroxide-producing activity of each strain showed a different pattern, and it was confirmed that the production of hydrogen peroxide actively occurred in Lactobacillus fermentum SNUV 175 , Lactobacillus jensenii SNUV 360, and Lactobacillus gasseri SNUV 281. Accordingly, the inhibitory activity of the selected strains is expected to show a difference in the mechanism of action.
- the acid resistance of the strains was determined by comparing the growth rate when cultured at 37° C. for 24 hours in acidic broth prepared by titrating MRS broth (pH 6.7) to pH 2 and pH 3, and the growth rate under basic broth conditions of pH 6.7, and the results are shown in Table 8 below.
- the growth rate of Lactobacillus isolates was measured after culturing in the media containing 0.1% to 4% of bile salt for 24 hours or more, and the results are shown in Table 9 and FIG. 1 .
- all four strains generally showed a growth rate of 50% or higher at the concentration of 0.1% bile salt.
- Lactobacillus fermentum SNUV 175 strain maintained the growth rate of 104.9% to 75.1%, compared to the non-treatment group at the concentration ranging from 0.5% to 4% of bile salt, showing a very high resistance to bile acid. Accordingly, it is expected to maintain a high survival rate even when orally administered.
- antibiotic resistance of the novel Lactobacillus sp. isolates having an activity to inhibit the growth of pathogenic vaginal microorganisms was evaluated.
- codes and standards concerning antibiotic resistance when utilizing Lactobacillus -based lactic acid bacteria in the food were not established, and thus it was evaluated based on the EFSA standard which is the international standard concerning antibiotic resistance of microorganisms added to animal feeds.
- the evaluation of the antibiotic resistance in Lactobacillus strains was performed according to the European Food Safety Authority (EFSA)'s guidelines for nine antibiotics including ampicillin (AMP), chloramphenicol (CHR), clindamycin (CLM), erythromycin (ERY), gentamycin KAN), streptomycin (STR), tetracycline (TET), vancomycin (VAN) and the like.
- EFSA European Food Safety Authority
- the test method used in the evaluation on the antibiotic resistance was performed according to ISO 10932:2010 (IDF 223: 2010), which is the SOP standard for antibiotic resistance test of lactic acid bacteria.
- Each Lactobacillus strain was inoculated at a density of ⁇ 6 ⁇ 10 6 CFU/mL in LSM-broth (90% IsoSensitest- and 10% MRS-broth; Oxoid), and then MIC test strip (Liofilchem, Italy) for each antibiotic was placed thereon. The degree of inhibition and MIC were evaluated after anaerobic culture at 37° C. for 48 hours, and the results are shown in Table 10 and FIG. 2
- Lactobacillus jensenii SNUV 360 and Lactobacillus fermentum SNUV 175 strains showed antibiotic susceptibility satisfying the EFSA criteria for all nine antibiotics used (EFSA guideline in Table 10), and thus it is expected to be used as a health functional food through oral administration
- the hormone control and estrous cycle were induced by intraperitoneally injecting 0.5 mg of beta-esteradiol 3-benzonate to female mice (BALB/c mice). After three days, they were directly infected with Gardnerella vaginalis in the vagina at a concentration of 1 ⁇ 10 7 CFU per mouse to establish a vaginitis animal model.
- Lactobacillus isolates corresponding to 10 8 to 10 9 CFU per mouse were vaginally administered ( 7 mice per group).
- the total bacterial DNA was extracted from the vaginal samples washed with 0.1 mL of PBS (Phosphate Buffered Saline) and microbiome community analysis was performed to measure the relative abundance of Gardnerella vaginalis and other vaginal microbiota.
- the DNA extraction from the vaginal fluid samples was performed using Mobio PowerSoil DNA extraction kit, and for the community analysis, the DNA was amplified via PCR using a primer corresponding to the V4 region of 16S rDNA, and next-generation sequencing analysis was carried out using Illumina Miseq equipment.
- the analyzed sequences were subjected to microbiome analysis including taxon profile, ⁇ -diversity and ⁇ -diversity showing the difference in community structure between groups through Qiime pipeline, and the change in the amount of Gardnerella vaginalis pathogens upon the administration of Lactobacillus isolates was evaluated by calculating relative abundances (/% GV treatment group). The results are shown in Table 11 and FIGS. 3 to 5 .
- Gardnerella vaginalis treatment group Lactobacillus crispatus SNUV 220 treatment group after infection with Gardnerella vaginalis, Lactobacillus jensenii SNUV 360 treatment group after infection with Gardnerella vaginalis, Lactobacillus fermentum SNUV 175 treatment group after infection with Gardnerella vaginalis, Lactobacillus gasseri SNUV 281 treatment group after infection with Gardnerella vaginalis , and metronidazole (0.75%) antibiotics treatment group as a positive control group after infection with Gardnerella vaginalis were designated as ‘GV’, ‘SNUV 220’, ‘SNUV 360’, ‘SNUV 175’, ‘SNUV 281’ and ‘MTZ’, respectively.
Abstract
Description
- The present invention activity relates to a Lactobacillus sp. strain having an ability to inhibit the growth of pathogenic vaginal microorganisms, a growth inhibitor of pathogenic vaginal microorganisms including the same as an active ingredient, and a product for improving, preventing or treating vaginitis including the inhibitor. More specifically, the present invention relates to a novel Lactobacillus sp. strain having an inhibitory effect on the growth of pathogenic vaginal microorganisms wherein the pathogenic vaginal microorganisms include fungi, bacteria and viruses, and specific examples thereof include Candidia spp. strains, Gardnerella spp. strains, or Sneathia spp. strains, or the like, and a pharmaceutical composition, a health functional food or a cleansing product including the same as an active ingredient.
- Globally, three-quarters of the female population is experiencing an infection of inflammatory diseases of the female genitalia including bacterial vaginitis during a period of their life, and it has been reported that 50% of them experience recurrence. In addition, more than 60% of female vaginitis corresponds to asymptomatic infections, and susceptibility to vaginitis is greatly increased through antibiotics, stress, hormonal changes after menopause, and the like.
- There are three types and causes of vaginitis caused by infection, including bacterial vaginosis (BV), candida vaginitis (CV) and trichomonas vulvovaginitis. Among them, the bacterial vaginosis has been recognized as the most common vaginitis, and it has been reported that Atopobium vaginae, Megasphaera sp., Gardnerella vaginalis, Eggerthella sp., Clostridium-like sp., Prevotella bivia, Peptostreptococcus micros, or the like have been found in the vaginal secretions isolated from patients with bacterial vaginosis. In particular, Gardnerella vaginalis is known as the major causative bacteria of bacterial vaginosis.
- Microbial communities in women's vagina are the major factor for maintaining vaginal health, and various bacteria, yeast and other microorganisms coexist in balance. The dominant species of healthy vaginal microflora is lactate-producing Lactobacillus sp., and it is known to inhibit various pathogens causing female diseases including vaginitis by performing functions, such as maintaining vaginal acidity, producing hydrogen peroxide and activating mucosal immune system, and the like.
- Conventional treatment of vaginitis using antibiotics is accompanied by various side effects, such as an increase in recurrence rate, a decrease in abundance of Lactobacillus spp., the generation of inflammation caused by other antibiotic resistant strains besides Gardnerella sp. and Candida sp. Therefore, the importance of inhibiting growth of vaginitis pathogens and recovering vaginal microflora using probiotic Lactobacillus spp. has been highlighted.
- Typical causative bacteria of vaginitis reported so far are Gardnerella vaginalis and Candida albicans, but recently, it has been reported that Sneathia spp. strains are closely associated with the infection of human papillomavirus and preterm birth, that they may also act as vaginitis pathogens in the disturbed vaginal microflora. Accordingly, there is a need to find Lactobacillus lactic acid bacteria having an inhibitory effect on the growth of Sneathia spp. pathogens, in addition to previously known pathogens for vaginitis.
- In the present invention, 190 strains of Lactobacillus spp. isolated from healthy women were screened for the inhibitory activity of pathogenic vaginal microorganisms. Probiotic Lactobacillus strains having novel activity have been found through the detection of Lactobacillus strains having an inhibitory effect against Sneathia spp. strains, which have been reported as microbiological markers associated with human papillomavirus infection and vaginitis-inducing causative bacteria, and a comprehensive inhibitory effect on the occurrence of vaginitis has been confirmed.
- Accordingly, it is one object of the present invention to provide a Lactobacillus sp. strain having an activity to inhibit the growth of pathogenic vaginal microorganisms.
- It is another object of the present invention to provide a pharmaceutical composition for treating and/or preventing diseases associated with the growth of pathogenic vaginal microorganisms, for example, vaginitis, containing the Lactobacillus sp. strain as an active ingredient.
- It is still another object of the present invention to provide a method for preventing and/or treating diseases associated with the growth of pathogenic vaginal microorganisms which comprises administering the Lactobacillus sp. strain to a patient having diseases associated with the growth of pathogenic vaginal microorganisms.
- It is further another object of the present invention to provide a health functional food or a cleansing product, etc., for improving, treating and/or preventing diseases associated with the growth of pathogenic vaginal microorganisms, for example, vaginitis, comprising the Lactobacillus sp. strain as an active ingredient.
- In order to achieve the objects above, the present invention provides a novel Lactobacillus sp. strain having an inhibitory effect on the growth of pathogenic vaginal microorganisms, and a pharmaceutical composition, a health functional food or a cleansing product comprising the same as an active ingredient.
- One embodiment of the present invention relates to a Lactobacillus sp. strain which is characterized by having an inhibitory activity on the growth of pathogenic vaginal microorganisms.
- The Lactobacillus sp. strain is isolated from the vaginal microflora of healthy women, and for example, it may be at least one Lactobacillus sp. strain selected from the group consisting of Lactobacillus crispatus SNUV 220, Lactobacillus fermentum SNUV 175, Lactobacillus jensenii SNUV 360, and Lactobacillus gasseri SNUV 281.
- The pathogenic vaginal microorganism may be at least one selected from the group consisting of fungi, bacteria and viruses. For example, it may be at least one selected from the group consisting of a Candida sp. strain being a causative bacteria of Candidal vaginitis, Sneathia spp. strains associated with the occurrence of bacterial vaginitis, a Gardnerella sp. strain, a vaginitis pathogen, and human papillomavirus associated with human papillomavirus infection. The Lactobacillus spp. strains may exhibit growth inhibitory effects against each of these strains or simultaneously and thus can be used for the prevention and/or treatment of female vaginitis.
- Moreover, the Candida sp. strain may be Candida albicans, but is not limited thereto. The Gardnerella sp. strain may be Gardnerella vaginalis, but is not limited thereto. The Sneathia spp. strain may be Sneathia amnii and Sneathia sanguinegens, but is not limited thereto.
- Further, the Lactobacillus sp. strain may function to actively produce hydrogen peroxide, and may also have acid resistance and bile resistance, but is not limited thereto.
- The hydrogen peroxide activity of the Lactobacillus sp. strain can be qualitatively discriminated for the strains that induces dark green color change, when cultured in TMB medium in which 3,3′, 5,5′-tetramethylbenzidine (250 mg/L), hemin (5 mg/L) and vitamin K (0.5 ug/L) are added to MRS agar medium for Lactobacillus culture.
- The acid resistance of the strains is determined based on the change in the growth rate when cultured in an acidic broth having a pH 2 and a pH 3 for 24 hours, and the bile resistance is determined based on the change in the growth rate when cultured for 24 hours in a broth supplemented with bile salt with a concentration ranging from 0.1% to 4% (w/v).
- The Lactobacillus sp. strain may exhibit a survival rate of 50% or higher at a concentration of 0.1% (w/v) bile salt, but is not limited thereto.
- Another embodiment of the present invention relates to a growth inhibitor of pathogenic vaginal microorganisms containing Lactobacillus sp. strain as an active ingredient, and a pharmaceutical composition for treating and/or preventing vaginitis, a food composition or cleansing product for improving, preventing or treating vaginitis including the inhibitor.
- The Lactobacillus sp. strain may be Lactobacillus crispatus, Lactobacillus fermentum, Lactobacillus jensenii, and Lactobacillus gasseri.
- Specifically, the strain may be at least one Lactobacillus sp. strain selected from the group consisting of Lactobacillus crispatus SNUV 220 deposited under accession No. KCTC18374P, Lactobacillus fermentum SNUV 175 deposited under accession No. KCTC18371P,
Lactobacillus jensenii SNUV 360 deposited under accession No. KCTC18372P,Lactobacillus gasseri SNUV 281 deposited under accession No. KCTC18375P - These strains were deposited with the Korean Collection for Type Cultures located in Yuseong-gu, Daejeon, South Korea on Apr. 7 and 9, 2015 and the accession numbers thereof were assigned.
- The growth inhibitor of vaginitis pathogens containing the Lactobacillus sp. strain according to the present invention as an active ingredient exhibits antiviral activity, antifungal activity against fungal pathogens, and antimicrobial activity against bacteria, and the vaginitis pathogens may be caused by at least one strain selected from the group consisting of a Candida sp. strain, a Gardnerella sp. strain, and a Sneathia spp. strain.
- For example, the Candida sp. strain may be Candida albicans, but is not limited thereto. Further, the Gardnerella sp. strain may be Gardnerella vaginalis, but is not limited thereto. Furthermore, the Sneathia spp. strain may be Sneathia amnii and Sneathia sanguinegens, but is not limited thereto
- The pharmaceutical composition according to the present invention can be administered to mammals including human via various routes. The mode of administration may be any mode commonly used in the art. For example, it may be administered by oral, transdermal, intravenous, intramuscular, subcutaneous routes or the like, and preferably, it may be orally administered.
- The pharmaceutical composition of the present invention may be used after being formulated into an oral preparation, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, etc., and a parental preparation, such as epidermal formulations, suppositories, or sterile injection solutions, in accordance with a conventional method
- The pharmaceutical composition of the present invention may further contain pharmaceutically suitable and physiologically acceptable adjuvants such as carriers, excipients and diluents, etc.
- Examples of carriers, excipients and diluents that can be included in the pharmaceutical composition of the present invention, may include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil. When formulated into a preparation, a diluting agent or an excipient, such as commonly-used fillers, weighting agents, binding agents, wetting agents, disintegrating agents, and surfactants can be used.
- When the pharmaceutical composition for preventing and/or treating vaginitis according to the present invention is provided for parenteral administration, for example, topical preparations such as liquid preparation, gel preparation, cleansing composition, tablet for vagal insertion, suppository, cream, ointment, dressing solution, spray, other coating agents, etc., liquid preparations such as solution-type, suspension-type, emulsion-type preparation, etc., and external skin formulations such as sterile aqueous solution, non-aqueous solvent, suspension, emulsion, freeze-drying preparation, suppository, cream, ointment, jelly, foam, cleansing agent or vaginal insert, preferably, liquid preparation, gel preparation, cleansing composition, tablet for vagal insertion, etc., may be included. The formulation can be prepared, for example, by adding a dissolution adjuvant, an emulsifying agent, a buffering agent for pH control, etc., to sterilized water.
- The non-aqueous solvents and suspending agents may include vegetable oils such as propylene glycol, polyethylene glycol and olive oil, and an injectable ester such as ethyl oleate and the like.
- In an embodiment where the pharmaceutical composition of the present invention is applied to humans, the pharmaceutical composition of the present invention may be administered alone, but considering the mode of administration and the standard pharmaceutical practice, it can be generally administered by mixing with the selected pharmaceutical carrier.
- For example, the composition containing the Lactobacillus sp. strain of the present invention may be orally, intrabuccally, or sublingually administered in a tablet form containing starch or lactose, in a capsule form containing only the active ingredient according to the present invention or containing an excipient in addition to the active ingredient, or in an elixir or suspension form containing a chemical agent for flavor or color.
- The dose of the pharmaceutical composition containing the Lactobacillus sp. strain of the present invention may vary depending on the patient's age, weight, sex, dosage form, health condition and severity of disease. In addition, it can be administered in divided doses once to several times a day at fixed time intervals according to the decision of a doctor or pharmacist. For example, the daily dose may be 0.1 to 500 mg/kg, preferably 0.5 to 300 mg/kg, based on the content of the active ingredient. The above doses are exemplified as an average case, and its dose may increase or decrease depending on individual differences. If the daily dose of the composition containing the mixture extract of the present invention is less than the above-mentioned dose, a significant effect cannot be obtained. If the daily dose exceeds the above-mentioned range, not only it is non-economical, but also it may cause undesirable side effects as the dose deviates from the above range.
- Still another embodiment of the present invention provides a health functional food for improving, treating and/or preventing vaginitis containing the Lactobacillus sp. strain as an active ingredient. The health functional food may be various beverages, fermented milk, food additives, and the like. The Lactobacillus sp. strain is as described above.
- The content of the Lactobacillus sp. strain as an effective ingredient contained in the health functional food is not particularly limited, but may appropriately vary depending on the form of food, desired use or the like. For example, it may be added in an amount of 0.01 to 15% by weight based on the total weight of the food, and the health beverage composition may be added at a ratio of 0.02 to 10 g, preferably 0.3 to 1 g, based on 100 ml.
- In the beverage among the health functional food of the present invention, there is no particular limitation on the liquid ingredient, except that the Lactobacillus strain is contained as an essential ingredient at the indicated ratio, and various flavoring agents or natural carbohydrates may be contained as additional ingredients as in common beverages.
- Examples of the above-mentioned natural carbohydrates include common sugars including monosaccharides, such as glucose, fructose, etc., disaccharides, such as maltose, sucrose, etc., and polysaccharides, such as dextrin, cyclodextrin, etc., as well as sugar alcohols, such as xylitol, sorbitol, erythritol, etc. As flavoring agents other than those mentioned above, natural flavoring agents (thaumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) may be favorably used. The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g, per 100 ml of the composition of the present invention.
- In addition to the above, the health functional food of the present invention may contain various nutrients, vitamins, minerals (electrolyte), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and flavor enhancers (cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickening agents, pH controlling agents, stabilizing agents, preservatives, glycerin, alcohol, carbonizing agents as used in carbonated beverages and the like.
- Moreover, the health functional food of the present invention may contain fruits, as used in preparing natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. Although the proportion of these additives is not of great importance, it is generally selected from a range of 0 to about 20 parts by weight per 100 parts by weight of the health functional food of the present invention.
- Still another embodiment of the present invention provides a cleansing product for improving, treating and/or preventing vaginitis containing the Lactobacillus sp. strain as an active ingredient. The cleansing product may be a solid cosmetic soap, a hand cleaner, a liquid shampoo, a liquid soap, a liquid conditioner, a body cleaner, a creamy soap, or the like. The Lactobacillus sp. strain is as described above.
- The cleansing product may further include a carrier for formulating a preparation. Examples of the carrier include a binding agent, a lubricant, an agent for suspension, a solubilizing agent, a buffer, a preservative, a lubricant, an isotonic agent, an excipient, a stabilizer, a dispersant, a suspending agent, a coloring agent, a flavoring agent, etc.
- The present invention relates to a novel Lactobacillus sp. strain having an ability to inhibit the growth of pathogenic vaginal microorganisms, and a pharmaceutical composition, a health functional food, and a cleansing product containing the same as an active ingredient. Therefore, the present invention exhibits an activity to inhibit the growth of Sneathia spp. pathogens associated with the human papillomavirus (HPV) infection and the occurrence of bacterial vaginitis, and an inhibitory effect on the growth of Gardnerella vaginalis which is a vaginitis pathogen, and Candida albicans which is a causative bacteria of Candidal vaginitis, and thus can be used for the improvement, prevention and/or treatment of female vaginitis.
-
FIG. 1 is a graph showing the measurement results of bile resistance by confirming the growth rate after culturing for 24 hours in a medium containing 0.1% to 4% (w/v) of bile salt according to one embodiment of the present invention. -
FIG. 2 is a representative graph showing the measurement results of antibiotic resistance of Lactobacillus fermentum SNUV 175 andLactobacillus jensenii SNUV 360 according to one embodiment of the present invention against nine antibiotics. -
FIG. 3 is a graph showing the relative abundance of Gardnerella vaginalis for each treatment group, which is the result of microbiome analysis after vaginal administration of four types of Lactobacillus strains according to one embodiment of the present invention into a mouse animal model infected with Gardnerella vaginalis. -
FIG. 4 is a principal component analysis (PCoA) graph showing the change in the community structure of vaginal microflora after 2 days of administration, which are the results of microbiome analysis after vaginal administration of four types of Lactobacillus strains according to one embodiment of the present invention into a mouse animal model infected with Gardnerella vaginalis. -
FIG. 5 is a graph showing the results of univariate analysis of microorganism groups with a significant change for each group, which are the results of microbiome analysis after vaginal administration of four types of Lactobacillus strains according to one embodiment of the present invention into a mouse animal model infected with Gardnerella vaginalis. - Hereinafter, the present invention will be described in more detail by way of Examples. However, these Examples are given for illustrative purposes only, and the scope of the invention is not intended to be limited by these Examples.
- About 190 different Lactobacillus strains were isolated from the vaginal microflora of healthy women. Specifically, the samples for the isolation of vaginal microflora were obtained from nine subjects, including three pairs of identical twins and their mothers who participated in the Korean Twin-Family Cohort Study, and were supplied from Samsung Hospital in the form of mid-vaginal swab samples (IRB No. 144-2011-07-11).
- The swab samples were transferred to the present laboratory in the form of being stored in a refrigerator within 4 hours after collection in a modified Liquid Amies solution and immediately used for the isolation of strains. The samples were sequentially diluted from 10−1 times to 10−8 times and spread on three different media of Chocolate agar, Rogosa agar, and Columbia agar, and cultured for 48 hours under anaerobic conditions.
- After the culture, purely isolated colonies were randomly selected and subjected to shake culture in brain heart infusion (BHI) broth containing 5% human serum. Genomic DNA was extracted from the cells, and PCR reaction was carried out using UnivFwd (5′-AGA GTT TGA TCM TGG CTC AG-3′) primer and UnivRev (5′-GGY TAC CTT GTT ACG ACT T-3′) primer for 16S ribosomal RNA typing. The PCR products were purified using QIAquick PCR purification kit and subjected to sequence analysis using ABI3711 automatic sequencer.
- The results are the same as those shown in Tables 1 and 2 below. Using such sequence information, the identification of the strains was finally completed by comparing with BLAST program of Genbank (www.ncbi.nlm.nhi.gov) and ExTaxon database program (www.ezbiocloud.net/eztaxon), together with the identification data of the previous result report.
-
TABLE 1 SEQ ID Species Name Nucleotide sequence (5′→3′) NO Lactobacillus SNUV TTACTTCGGCAATGACGTTAGGAAAGC 1 crispatus 220 GAGCGGCGGATGGGTGAGTAACACGTG GGGAACCTGCCCCATAGTCTGGGATAC CACTTGGAAACAGGTGCTAATACCGGA TAAGAAAGCAGATCGCATGATCAGCTT TTAAAAGGCGGCGTAAGCTGTCGCTAT GGGATGGCCCCGCGGTGCATTAGCTAG TTGGTAAGGTAAAGGCTTACCAAGGCG ATGATGCATAGCCGAGTTGAGAGACTG ATCGGCCACATTGGGACTGAGACACGG CCCAAACTCCTACGGGAGGCAGCAGTA GGGAATCTTCCACAATGGACGCAAGTC TGATGGAGCAACGCCGCGTGAGTGAAG AAGGTTTTCGGATCGTAAAGCTCTGTT GTTGGTGAAGAAGGATAGAGGTAGTAA CTGGCCTTTATTTGACGGTAATCAACC AGAAAGTCACGGCTAACTACGTGCCAG CAGCCGCGGTAATACGTAGGTGGCAAG CGTTGTCCGGATTTATTGGGCGTAAGC GAGCGCAGGCGGAAGAATAAGTCTGAT GTGAAAGCCCTCGGCTTAACCGAGGAA CTGCATCGGAAACTGTTTTTCTTGAGT GCAGAAGAGGAGAGTGGAACTCCATGT GTAGCGGTGGAATGCGTAGATATATGG AAGAACACCAGTGGCGAAGGCGGCTCT CTGGTCTGCAACTGACGCTGAGGCTCG AAAGCATGGGTAGCGAACAGGATTAGA TACCCTGGTAGTCCATGCCGTAAACGA TGAGTGCTAAGTGTTGGGAGGTTTCCG CCTCTCAGTGCTGCAGCTAACGCATTA AGCACTCCGCCTGGGGAGTACGACCGC AAGGTTGAAACTCAAAGGAATTGACGG GGGCCCGCACAAGCGGTGGAGCATGTG GTTTAATTCGAAGCAACGCGAAGAACC TTACCAGGTCTTGACATCTAGTGCC Lactobacillus SNUV CTGCCCAGAAGCGGGGGACAACATTTG 2 fermentum 175 GAAACAGATGCTAATACCGCATAACAA CGTTGTTCGCATGAACAACGCTTAAAA GATGGCTTCTCGCTATCACTTCTGGAT GGACCTGCGGTGCATTAGCTTGTTGGT GGGGTAACGGCCTACCAAGGCGATGAT GCATAGCCGAGTTGAGAGACTGATCGG CCACAATGGGACTGAGACACGGCCCAT ACTCCTACGGGAGGCAGCAGTAGGGAA TCTTCCACAATGGGCGCAAGCCTGATG GAGCAACACCGCGTGAGTGAAGAAGGG TTTCGGCTCGTAAAGCTCTGTTGTTAA AGAAGAACACGTATGAGAGTAACTGTT CATACGTTGACGGTATTTAACCAGAAA GTCACGGCTAACTACGTGCCAGCAGCC GCGGTAATACGTAGGTGGCAAGCGTTA TCCGGATTTATTGGGCGTAAAGAGAGT GCAGGCGGTTTTCTAAGTCTGATGTGA AAGCCTTCGGCTTAACCGGAGAAGTGC ATCGGAAACTGGATAACTTGAGTGCAG AAGAGGGTAGTGGAACTCCATGTGTAG CGGTGGAATGCGTAGATATATGGAAGA ACACCAGTGGCGAAGGCGGCTACCTGG TCTGCAACTGACGCTGAGACTCGAAAG CATGGGTAGCGAACAGGATTAGATACC CTGGTAGTCCATGCCGTAAACGATGAG TGCTAGGTGTTGG -
TABLE 2 SEQ ID Species Name Nucleotide sequence (5′→3′) NO Lactobacillus SNUV AAAAGCTACTTTCGCATGAAAGAAGTT 3 jensenii 360 TAAAAGGCGGCGTAAGCTGTCGCTAAA GGATGGACCTGCGATGCATTAGCTAGT TGGTAAGGTAACGGCTTACCAAGGCGA TGATGCATAGCCGAGTTGAGAGACTGA TCGGCCACATTGGGACTGAGACACGGC CCAAACTCCTACGGGAGGCAGCAGTAG GGAATCTTCCACAATGGACGAAAGTCT GATGGAGCAACGCCGCGTGAGTGAAGA AGGTTTTCGGATCGTAAAGCTCTGTTG TTGGTGAAGAAGGATAGAGGTAGTAAC TGGCCTTTATTTGACGGTAATCAACCA GAAAGTCACGGCTAACTACGTGCCAGC AGCCGCGGTAATACGTAGGTGGCAAGC GTTGTCCGGATTTATTGGGCGTAAAGC GAGCGCAGGCGGATTGATAAGTCTGAT GTGAAAGCCTTCGGCTCAACCGAAGAA CTGCATCAGAAACTGTCAATCTTGAGT GCAGAAGAGGAGAGTGGAACTCCATGT GTAGCGGTGGAATGCGTAGATATATGG AAGAACACCAGTGGCGAAGGCGGCTCT CTGGTCTGTAACTGACGCTGAGGCTCG AAAGCATGGGTAGCGAACAGGATTAGA TACCCTGGTAGTCCATGCCGTAAACGA TGAGTGCTAAGTGTTGGGAGGTTTCCG CCTCTCAGTGCTGCAGCTAACGCATTA AGCACTCCGCCTGGGG Lactobacillus SNUV CGGATAACAACACTAGACGCATGTCTA 4 gasseri 281 GAGTTTAAAAGATGGTTCTGCTATCAC TCTTGGATGGACCTGCGGTGCATTAGC TAGTTGGTAAGGCAACGGCTTACCAAG GCAATGATGCATAGCCGAGTTGAGAGA CTGATCGGCCACATTGGGACTGAGACA CGGCCCAAACTCCTACGGGAGGCAGCA GTAGGGAATCTTCCACAATGGACGCAA GTCTGATGGAGCAACGCCGCGTGAGTG AAGAAGGGTTTCGGCTCGTAAAGCTCT GTTGGTAGTGAAGAAAGATAGAGGTAG TAACTGGCCTTTATTTGACGGTAATTA CTTAGAAAGTCACGGCTAACTACGTGC CAGCAGCCGCGGTAATACGTAGGTGGC AAGCGTTGTCCGGATTTATTGGGCGTA AAGCGAGTGCAGGCGGTTCAATAAGTC TGATGTGAAAGCCTTCGGCTCAACCGG GAATTGCATCAGAAACTGTTGAACTTG AGTGCAGAAGAGGAGAGTGGAACTCCA TGTGTAGCGGTGGAATGCGTAGATATA TGGAAGAACACCAGTGGCGAAGGCGGC TCTCTGGTCTGCAACTGACGCTGAGGC TCGAAAGCATGGGTAGCGAACAGGATT AGATACCCTGGTAGTCCATGCCGTAAA CGATGAGTGCTAAGTGTTGGGAGGTTT CCGCCTCTCAGTGCTGCAGCTAACGCA TTAAGCACTCCGCCTGGGG -
TABLE 3 Pairwise Diff/ Similarity Total Completeness Name Taxonomy Accession (%) nt (%) SNUV Bacteria; Firmicutes; Bacilli; Y17362 100 0/650 100 220 Lactobacillales; Lactobacillaceae; Lactobacillus; Lactobacillus crispatus; SNUV Bacteria; Firmicutes; Bacilli; AP008937 99.86 1/715 100 175 Lactobacillales; Lactobacillaceae; Lactobacillus; Lactobacillus fermentum; Bacteria; Firmicutes; Bacilli; AJ575812 99.86 1/715 100 Lactobacillales; Lactobacillaceae; Lactobacillus; Lactobacillus fermentum; SNUV Bacteria; Firmicutes; Bacilli; AF243176 99.86 1/718 98.9 360 Lactobacillales; Lactobacillaceae; Lactobacillus; Lactobacillus jensenii; Bacteria; Firmicutes; Bacilli; Y18654 99.86 1/717 95.5 Lactobacillales; Lactobacillaceae; Lactobacillus; Lactobacillus fornicalis; SNUV Bacteria; Firmicutes; Bacilli; CP000413 99.86 1/722 100 281 Lactobacillales; Lactobacillaceae; Lactobacillus; Lactobacillus gasseri; - As shown in Tables 1 to 3, the results were found to be Lactobacillus crispatus, Lactobacillus fermentum, Lactobacillus jensenii, and Lactobacillus gasseri, respectively. Accordingly, they were assigned SNUV 220, SNUV 175,
SNUV 360, andSNUV 281, respectively, and deposited with the Korean Collection for Type Cultures (KCTC) located in Yuseong-gu, Daejeon, South Korea and were assigned accession numbers KCTC18374P (SNUV 220, deposited on April 9), KCTC18371P (SNUV 175, deposited on April 7), KCTC18372P (SNUV 360, deposited on April 7) and KCTC18375P (SNUV 281, deposited on April 9). - For pure isolation and long-term storage of the identified strains, glycerol (16% v/v) was added to the culture medium that reached the exponential phase, and stored at −80° C. as a stock. In order to prepare cell culture medium for the evaluation of inhibitory activity of vaginitis pathogens for each strain, 1% of each strain was inoculated in an anaerobic MRS medium and cultured at 37° C. for 24 hours. Then, the microbial cells were removed by centrifugation at 13000×g for 10 minutes, and the supernatant was passed through a membrane filter with a pore size of 0.22 um and then stored at −80° C. until they were used in the experiment.
- Each of Lactobacillus strains isolated in Example 1 was used for the assay by culturing at 37° C. for 20 hours using MRS broth (Difco, USA). The strains of Sneathia spp. and Gardnerella vaginalis were inoculated into NYCIII broth and then anaerobically cultured at 37° C. for 48 hours or 24 hours, respectively, and used in the experiment.
- As for the Sneathia spp. strains used in the experiment, two kinds of strains of Sneathia spp., which was isolated in Virginia Commonwealth University School of Medicine and has been reported in the paper, and Sneathia sanguinegens that the present inventors isolated from Korean women were used. As the Gardnerella vaginalis strain used in the experiment, KCTC 5096 strain furnished from the Korean Collection for Type Cultures (KCTC) was used.
- After dispensing and solidifying 15 ml of MRS agar medium on a plate, 7 ml of NYCIII soft agar medium (0.75% agar) inoculated with the strains of Sneathia spp. and Gardnerella vaginalis at a density of 5×106 CFU/mL was formed as multilayer medium. Once the upper soft agar medium had been solidified, a diffusion paper disc (8 mm in diameter) was placed on the medium, and about 20 μl of culture product of each Lactobacillus strain was absorbed thereto. Then, the plate was added to an anaerobic jar and anaerobically incubated at 37° C. for 48 hours. After the incubation, the growth inhibitory zone of the strains of Sneathia spp. and Gardnerella vaginalis appearing around the disk was measured. The inhibitory activity was expressed as the diameter (mm) of the transparent disk in which the growth of the strains was inhibited from the center of the disk.
- The disk inhibition assay was performed against the strains of Sneathia spp. and Gardnerella vaginalis with culture supernatant of isolated Lactobacillus strains, and the representative result are shown in Table 4 below.
-
TABLE 4 disk inhibition (diameter, mm) Sn. Species Isolates_No Sn. Amnii Sanguinegens G. vaginalis Lactobacillus SNUV 220 23 46 19 crispatus Lactobacillus SNUV 175 — 19 — fermentum Lactobacillus SNUV 360 60 60 25 jensenii Lactobacillus SNUV 281 — 23 19 gasseri Lactobacillus SNUV 215 — — — crispatus Lactobacillus SNUV 110 — — — fermentum Lactobacillus SNUV 212 — — — jensenii Lactobacillus SNUV 445 — — — gasseri - As can be confirmed in Table 4, although the size of disk inhibition of the Lactobacillus culture supernatant was slightly different depending on the type of the inhibitory strains, four types of Lactobacillus isolated strains which simultaneously inhibited the growth of Sneathia spp. and Gardnerella vaginalis strains, that is, Lactobacillus crispatus SNUV 220, Lactobacillus fermentum SNUV 175,
Lactobacillus jensenii SNUV 360, andLactobacillus gasseri SNUV 281 were selected. In particular, theLactobacillus jensenii SNUV 360 strain exhibited a most potent inhibitory activity against both strains of Sneathia spp. and Gardnerella sp. - In order to confirm the inhibitory activity against Candida albicans strain, which is a causative bacteria of Candidal vaginitis of the four kinds of Lactobacillus strains screened in Example 2 above, Candida albicans ATCC44858 strain (American Type Culture Collection), and MYA4788 strain (American Type Culture Collection), which has been proven to cause vaginitis in animal experiments, were selected as target strains for selection of Lactobacillus isolates having an inhibitory function, and the experiments was conducted accordingly.
- Specifically, the Candida albicans strains were evaluated using a 96-well diffusion test. For the experiment, 50 ul of Candida albicans ATCC44858 (or Candida albicans M4788) culture media diluted by adding 100 ul of YM medium and 100 ul of Lactobacillus culture supernatant was added to a well of 96-well plate, and then cultured at 37° C. for 24 hours. Thereafter, the growth of inhibited Candida strains was estimated by measurement of absorbance at 600 nm, and the results are shown in Table 5.
-
TABLE 5 optical density after 24 h (O.D. at 600 nm) Candida Candida 44858 MYA4788 Species Isolates_No (co-culture) (co-culture) Negative control group 1 1 Lactobacillus crispatus SNUV 220 0.028 0.007 Lactobacillus SNUV 175 — 0.003 fermentum Lactobacillus jensenii SNUV 360 — 0.007 Lactobacillus gasseri SNUV 281 0.012 0.010 - As can be confirmed in Table 5, all the selected Lactobacillus crispatus SNUV 220, Lactobacillus fermentum SNUV 175,
Lactobacillus jensenii SNUV 360, andLactobacillus gasseri SNUV 281 had a killing activity against Candida strains close to 100%. Accordingly, all four isolated strains have been shown to have a significant effect on the prevention of vaginitis. - In order to investigate the degree of hydrogen peroxide production, a TMB agar medium was prepared as shown in Table 6 below (medium composition per 1 L).
-
TABLE 6 Difco Lactobacilli MRS medium 55 g TMB 250 mg Starch, soluble 100 g Hemnin solution 10 mL Vitamin K solution 0.2 mL Peroxidase solution (1 mg/mL) 10 mL - Subsequently, each of Lactobacillus crispatus SNUV 220, Lactobacillus fermentum SNUV 175,
Lactobacillus jensenii SNUV 360, andLactobacillus gasseri SNUV 281, which are the four types of Lactobacillus strains screened in Example 2, was cultured in MRS broth at 37° C. for 20 hours, inoculated onto the TMB agar plate, and then anaerobically cultured at 37° C. for 2 days. After the culture, the plate was exposed to the air for 30 minutes and evaluated via a qualitative experiment in which the color of the colonies turns to blue. The extent to which the color of colonies turned to blue was observed with the naked eye, and the results are shown in Table 7 below. -
TABLE 7 Species Isolates_No H2O2 production Lactobacillus crispatus SNUV 220 − Lactobacillus fermentum SNUV 175 +++ Lactobacillus jensenii SNUV 360 +++ Lactobacillus gasseri SNUV 281 ++ - As can be confirmed in Table 7, the hydrogen peroxide-producing activity of each strain showed a different pattern, and it was confirmed that the production of hydrogen peroxide actively occurred in Lactobacillus fermentum SNUV 175,
Lactobacillus jensenii SNUV 360, andLactobacillus gasseri SNUV 281. Accordingly, the inhibitory activity of the selected strains is expected to show a difference in the mechanism of action. - The acid resistance of the strains was determined by comparing the growth rate when cultured at 37° C. for 24 hours in acidic broth prepared by titrating MRS broth (pH 6.7) to pH 2 and pH 3, and the growth rate under basic broth conditions of pH 6.7, and the results are shown in Table 8 below.
-
TABLE 8 Species Isolates_No pH 6.7 pH 3 pH 2 Lactobacillus SNUV 220 +++ + − crispatus Lactobacillus SNUV 175 +++ +++ + fermentum Lactobacillus SNUV 360 +++ + − jensenii Lactobacillus SNUV 281 ++ + + gasseri - As can be confirmed in Table 8, all four strains showed the growth under the culture conditions of pH 3, and in particular, SNUV 175 strain and
SNUV 281 strain showed the growth even under the condition of pH 2, indicating that they have a strong acid resistance. - For the evaluation of bile resistance, the growth rate of Lactobacillus isolates was measured after culturing in the media containing 0.1% to 4% of bile salt for 24 hours or more, and the results are shown in Table 9 and
FIG. 1 . -
TABLE 9 bile salts bile salts bile salts bile salts bile salts Species 4% 2% 1% 0.5% 0.1% Negative control group 100 100 100 100 100 Lactobacillus crispatus 2.1 13.7 12.1 27.0 81.8 SNUV 220 Lactobacillus fermentum 75.1 88.0 96.5 104.9 127.1 SNUV 175 Lactobacillus jensenii SNUV 13.5 12.6 22.8 24.1 59.3 360 Lactobacillus gasseri SNUV 2.2 16.2 15.3 24.4 76.9 281 - As can be confirmed in Table 9 and
FIG. 1 , all four strains generally showed a growth rate of 50% or higher at the concentration of 0.1% bile salt. In particular, Lactobacillus fermentum SNUV 175 strain maintained the growth rate of 104.9% to 75.1%, compared to the non-treatment group at the concentration ranging from 0.5% to 4% of bile salt, showing a very high resistance to bile acid. Accordingly, it is expected to maintain a high survival rate even when orally administered. - In order to confirm the safety when applied to functional food materials, etc., antibiotic resistance of the novel Lactobacillus sp. isolates having an activity to inhibit the growth of pathogenic vaginal microorganisms was evaluated. Currently, codes and standards concerning antibiotic resistance when utilizing Lactobacillus-based lactic acid bacteria in the food were not established, and thus it was evaluated based on the EFSA standard which is the international standard concerning antibiotic resistance of microorganisms added to animal feeds.
- Specifically, the evaluation of the antibiotic resistance in Lactobacillus strains was performed according to the European Food Safety Authority (EFSA)'s guidelines for nine antibiotics including ampicillin (AMP), chloramphenicol (CHR), clindamycin (CLM), erythromycin (ERY), gentamycin KAN), streptomycin (STR), tetracycline (TET), vancomycin (VAN) and the like. The test method used in the evaluation on the antibiotic resistance was performed according to ISO 10932:2010 (IDF 223: 2010), which is the SOP standard for antibiotic resistance test of lactic acid bacteria. Each Lactobacillus strain was inoculated at a density of ˜6×106 CFU/mL in LSM-broth (90% IsoSensitest- and 10% MRS-broth; Oxoid), and then MIC test strip (Liofilchem, Italy) for each antibiotic was placed thereon. The degree of inhibition and MIC were evaluated after anaerobic culture at 37° C. for 48 hours, and the results are shown in Table 10 and
FIG. 2 -
TABLE 10 L. jensenii SNUV EFSA L. fermentum EFSA antibiotics 360 guideline SNUV 175 guideline AMP 0.094 1 0.125 2 CHL 3 4 2 4 CLM 0.38 1 0.064 1 ERY 0.19 1 0.38 1 GEN 2 16 1 16 KAN 4 16 24 32 STR 3 16 12 64 TET 0.75 4 2 8 VAN 0.5 2 64 Not required - As can be confirmed in Table 10 and
FIG. 2 , among the four isolation strains,Lactobacillus jensenii SNUV 360 and Lactobacillus fermentum SNUV 175 strains showed antibiotic susceptibility satisfying the EFSA criteria for all nine antibiotics used (EFSA guideline in Table 10), and thus it is expected to be used as a health functional food through oral administration - The hormone control and estrous cycle were induced by intraperitoneally injecting 0.5 mg of beta-esteradiol 3-benzonate to female mice (BALB/c mice). After three days, they were directly infected with Gardnerella vaginalis in the vagina at a concentration of 1×107 CFU per mouse to establish a vaginitis animal model.
- Thereafter, four kinds of Lactobacillus isolates corresponding to 108 to 109 CFU per mouse were vaginally administered (7 mice per group). On day 2, the total bacterial DNA was extracted from the vaginal samples washed with 0.1 mL of PBS (Phosphate Buffered Saline) and microbiome community analysis was performed to measure the relative abundance of Gardnerella vaginalis and other vaginal microbiota. The DNA extraction from the vaginal fluid samples was performed using Mobio PowerSoil DNA extraction kit, and for the community analysis, the DNA was amplified via PCR using a primer corresponding to the V4 region of 16S rDNA, and next-generation sequencing analysis was carried out using Illumina Miseq equipment.
- The analyzed sequences were subjected to microbiome analysis including taxon profile, α-diversity and β-diversity showing the difference in community structure between groups through Qiime pipeline, and the change in the amount of Gardnerella vaginalis pathogens upon the administration of Lactobacillus isolates was evaluated by calculating relative abundances (/% GV treatment group). The results are shown in Table 11 and
FIGS. 3 to 5 . - Gardnerella vaginalis treatment group, Lactobacillus crispatus SNUV 220 treatment group after infection with Gardnerella vaginalis,
Lactobacillus jensenii SNUV 360 treatment group after infection with Gardnerella vaginalis, Lactobacillus fermentum SNUV 175 treatment group after infection with Gardnerella vaginalis,Lactobacillus gasseri SNUV 281 treatment group after infection with Gardnerella vaginalis, and metronidazole (0.75%) antibiotics treatment group as a positive control group after infection with Gardnerella vaginalis were designated as ‘GV’, ‘SNUV 220’, ‘SNUV 360’, ‘SNUV 175’, ‘SNUV 281’ and ‘MTZ’, respectively. -
TABLE 11 Relative abundance (/% GV group) GV 100 SNUV 220 18.0 SNUV 360 5.3 SNUV 175 2.6 SNUV 281 6.9 MTZ 74.9 - As can be confirmed in Table 11 and
FIG. 3 , the strains of Lactobacillus crispatus SNUV 220,Lactobacillus jensenii SNUV 360, Lactobacillus fermentum SNUV 175, Lactobacillus gasseri of the present invention have been found to reduce the amount of Gardnerella vaginalis pathogens in in vivo vaginitis model. - As can be confirmed in
FIG. 4 , when the four Lactobacillus strains of the present invention were administered, it was found that the community structure of the vaginal microflora was all changed compared to Gardnerella vagina/is-infected control group. The changes in the community structure after administration of Gardnerella and Lactobacillus strains were measured using Unweighted UniFrac distance and are shown inFIG. 4 . During the vaginal administration of metronidazole, a positive control currently used as a therapeutic agent for vaginitis disease, there was no change in the community structure observed in the treatment group of Lactobacillus strains. - In addition to confirming that the microbial community structure in the vagina, significantly changed taxon profile was analyzed by LefSe program after administration of Lactobacillus strains to Gardnerella vagina/is-infected mouse.
- The results are as shown in cladogram of
FIG. 5 . From the above analysis, it was confirmed that Gardnerella vaginalis and Staphylococcus spp. were significantly increased in group (C) infected with Gardnerella alone, and the Lactobacillus strains were significantly increased in groups (D, E, G) in which the Lactobacillus strains of the present invention were administered once after infection with Gardnerella vaginalis, thereby the administration of Lactobacillus strains changed the vaginal community structure and taxa profile and had a modulatory effect on the vaginal microflora.
Claims (25)
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR10-2015-0053702 | 2015-04-16 | ||
KR20150053702 | 2015-04-16 | ||
KR1020160022840A KR101860552B1 (en) | 2015-04-16 | 2016-02-25 | Lactobacilli having inhibitory effect against pathogenic microorganisms in vagina |
KR10-2016-0022840 | 2016-02-25 | ||
PCT/KR2016/002988 WO2016167489A1 (en) | 2015-04-16 | 2016-03-24 | Lactobacillus sp. strain having ability to inhibit proliferation of virginal pathogenic microorganisms |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/KR2016/002988 A-371-Of-International WO2016167489A1 (en) | 2015-04-16 | 2016-03-24 | Lactobacillus sp. strain having ability to inhibit proliferation of virginal pathogenic microorganisms |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US17/019,677 Continuation US20200405789A1 (en) | 2015-04-16 | 2020-09-14 | Lactobacillus sp. strain having ability to inhibit growth of virginal pathogenic microorganisms |
Publications (1)
Publication Number | Publication Date |
---|---|
US20180117100A1 true US20180117100A1 (en) | 2018-05-03 |
Family
ID=57251933
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US15/566,352 Abandoned US20180117100A1 (en) | 2015-04-16 | 2016-03-24 | Lactobacillus sp. strain having ability to inhibit proliferation of virginal pathogenic microorganisms |
US17/019,677 Pending US20200405789A1 (en) | 2015-04-16 | 2020-09-14 | Lactobacillus sp. strain having ability to inhibit growth of virginal pathogenic microorganisms |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US17/019,677 Pending US20200405789A1 (en) | 2015-04-16 | 2020-09-14 | Lactobacillus sp. strain having ability to inhibit growth of virginal pathogenic microorganisms |
Country Status (2)
Country | Link |
---|---|
US (2) | US20180117100A1 (en) |
KR (3) | KR101860552B1 (en) |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111135157A (en) * | 2018-07-19 | 2020-05-12 | 大江生医股份有限公司 | Use of a combination of 3-phenyllactic acid and prebiotics for improving the bacterial phase |
JP2021524747A (en) * | 2018-05-23 | 2021-09-16 | コ・バイオラブズ・インコーポレイテッド | Lactobacillus crispatus KBL693 strain and its use |
CN114129601A (en) * | 2021-12-22 | 2022-03-04 | 南方医科大学第七附属医院(佛山市南海区第三人民医院) | Application of lactobacillus gasseri LGV03 in preparation of medicine for preventing or treating HPV infection |
JP2022524157A (en) * | 2019-08-09 | 2022-04-27 | スーチュァン アナエロビック バイオテクノロジー カンパニー リミテッド | Composite Lactobacillus Composition and Its Uses in Vaginal Health in Women |
CN114452307A (en) * | 2020-11-10 | 2022-05-10 | 西安正浩生物制药有限公司 | Lactobacillus capsule with treatment and prevention effects on vaginitis and preparation method thereof |
Families Citing this family (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR102063757B1 (en) * | 2019-05-03 | 2020-01-08 | 주식회사 메디오젠 | Lactobacillus for the treatment or improvement of vaginitis with antimicrobial effect on Gardnerella vaginalis and Candida albicans |
KR102269965B1 (en) * | 2020-12-31 | 2021-06-28 | 주식회사 바이오뱅크힐링 | Lactobacillus paragasseri strain, and vesicles from thereof and anti-inflammation and anti-bacteria uses of thereof |
KR102296289B1 (en) * | 2020-12-31 | 2021-09-01 | 주식회사 바이오뱅크힐링 | Lactobacillus gasseri strain, and vesicles from thereof and anti-inflammation and anti-bacteria uses of thereof |
KR102488955B1 (en) * | 2022-01-14 | 2023-01-17 | 주식회사 메디오젠 | Feminine cleanser with skin moisturizing ability |
KR102434486B1 (en) * | 2022-04-06 | 2022-08-19 | 주식회사 큐옴바이오 | Vagina-derived lactobacillus rhamnosus vg.q1 strain having antimicrobial activity against microorganisms causing vaginosis and uses thereof |
KR102434487B1 (en) * | 2022-04-06 | 2022-08-19 | 주식회사 큐옴바이오 | Vagina-derived lactobacillus rhamnosus vg.q2 strain having antimicrobial activity against microorganisms causing vaginosis and uses thereof |
KR102554125B1 (en) * | 2022-10-24 | 2023-07-13 | (주)헥토헬스케어 | Lactobacillus gasseri BELG08 preventive or ameliorating effect on vaginitis and composition comprising the same |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1424075A2 (en) * | 2002-10-30 | 2004-06-02 | Food Industry Research and Development Institute | Acid and bile-resistant lactobacillus isolates having the ability to lower and addimilate cholesterol |
US20140147425A1 (en) * | 2012-11-23 | 2014-05-29 | Seres Health, Inc. | Synergistic bacterial compositions and methods of production and use thereof |
-
2016
- 2016-02-25 KR KR1020160022840A patent/KR101860552B1/en active IP Right Grant
- 2016-03-24 US US15/566,352 patent/US20180117100A1/en not_active Abandoned
-
2017
- 2017-03-31 KR KR1020170041922A patent/KR101867048B1/en active IP Right Grant
-
2018
- 2018-01-10 KR KR1020180003582A patent/KR101860553B1/en active IP Right Grant
-
2020
- 2020-09-14 US US17/019,677 patent/US20200405789A1/en active Pending
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1424075A2 (en) * | 2002-10-30 | 2004-06-02 | Food Industry Research and Development Institute | Acid and bile-resistant lactobacillus isolates having the ability to lower and addimilate cholesterol |
US20140147425A1 (en) * | 2012-11-23 | 2014-05-29 | Seres Health, Inc. | Synergistic bacterial compositions and methods of production and use thereof |
Non-Patent Citations (1)
Title |
---|
Kim Asian-Australian Journal of Animal Science, Vol. 19, no9, pp.1335-1341; 2006 * |
Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2021524747A (en) * | 2018-05-23 | 2021-09-16 | コ・バイオラブズ・インコーポレイテッド | Lactobacillus crispatus KBL693 strain and its use |
US11633439B2 (en) | 2018-05-23 | 2023-04-25 | Kobiolabs, Inc. | Lactobacillus crispatus KBL693 strain and use thereof |
JP7442195B2 (en) | 2018-05-23 | 2024-03-04 | コバイオラブズ・インコーポレイテッド | Lactobacillus crispatus KBL693 strain and its use |
CN111135157A (en) * | 2018-07-19 | 2020-05-12 | 大江生医股份有限公司 | Use of a combination of 3-phenyllactic acid and prebiotics for improving the bacterial phase |
JP2022524157A (en) * | 2019-08-09 | 2022-04-27 | スーチュァン アナエロビック バイオテクノロジー カンパニー リミテッド | Composite Lactobacillus Composition and Its Uses in Vaginal Health in Women |
JP7354274B2 (en) | 2019-08-09 | 2023-10-02 | スーチュァン アナエロビック バイオテクノロジー カンパニー リミテッド | Complex lactobacillus composition and its use in female vaginal health |
CN114452307A (en) * | 2020-11-10 | 2022-05-10 | 西安正浩生物制药有限公司 | Lactobacillus capsule with treatment and prevention effects on vaginitis and preparation method thereof |
CN114129601A (en) * | 2021-12-22 | 2022-03-04 | 南方医科大学第七附属医院(佛山市南海区第三人民医院) | Application of lactobacillus gasseri LGV03 in preparation of medicine for preventing or treating HPV infection |
Also Published As
Publication number | Publication date |
---|---|
KR20180006998A (en) | 2018-01-19 |
KR101860553B1 (en) | 2018-05-23 |
KR20160123983A (en) | 2016-10-26 |
KR101867048B1 (en) | 2018-06-12 |
KR20170039637A (en) | 2017-04-11 |
KR101860552B1 (en) | 2018-07-02 |
US20200405789A1 (en) | 2020-12-31 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20200405789A1 (en) | Lactobacillus sp. strain having ability to inhibit growth of virginal pathogenic microorganisms | |
JP4455333B2 (en) | Probiotic bacteria: Lactobacillus fermentum | |
US8846027B2 (en) | Compositions for the vaginal and oral administration of Lactobacillus and uses thereof | |
CN1888051B (en) | Plant lactobacillus strain and its application | |
JP6837015B2 (en) | For example, Lactobacillus rhamnosus bacteria for the treatment of bacterial vaginosis | |
KR101638984B1 (en) | Nano-Sized Lactic Acid Bacteria from Kimchi | |
TWI627276B (en) | Novel lactobacillus crispatus strain | |
KR101287126B1 (en) | Pharmaceutical compositon prevention and treatment of vaginitis and urinary tract infection comprising fermented solution of plant-originated Lactic acid bacteria | |
AU2021269282B2 (en) | Novel lactic acid bacteria and use thereof | |
KR102323783B1 (en) | Clostridium scindens having inhibitory effect against Clostridium difficile | |
KR102084973B1 (en) | Composition for preventing or treating colitis comprising enterococcus faecalis | |
KR102543494B1 (en) | Novel probiotics and use thereof | |
EP3261723B1 (en) | Probiotic lactobacillus plantarum strains for urinary tract infections | |
KR101957211B1 (en) | Lactobacillus sp. strain having antimicrobial activity against microorganisms causing premature birth and vaginosis, antiviral activity againt HSV and improved vagina adhesion ability and uses thereof | |
WO2018112741A1 (en) | Lactobacillus acidophilus, culture method therefor and application thereof | |
KR102434487B1 (en) | Vagina-derived lactobacillus rhamnosus vg.q2 strain having antimicrobial activity against microorganisms causing vaginosis and uses thereof | |
CN114774315B (en) | Application of lactobacillus rhamnosus strain LRa05 in preparation of immunity enhancing product and/or eczema relieving product | |
KR102486970B1 (en) | Lactobacillus plantarum having inhibitory effect against Clostridium difficile | |
US20130309212A1 (en) | Lactobacillus salivarius and method for preparing metabolite thereof, composition of lactobacillus salivarius and metabolite thereof and use of the composition | |
KR20200018532A (en) | Lactobacillus sp. strain having inhibitory effect against microorganisms causing vaginosis uses thereof | |
KR101355441B1 (en) | Lactobacillus johnsonii HY7042 helpful to maintain healthy vaginal environment and products containing thereof as effective component | |
KR101960189B1 (en) | NOVEL STRAIN OF Lactobacillus plantarum AND USE THEREOF | |
EP3932416A2 (en) | Composition for improving, preventing, or treating bone diseases or metabolic diseases, including novel lactobacillus sakei cvl-001 strain and culture medium thereof | |
KR102434486B1 (en) | Vagina-derived lactobacillus rhamnosus vg.q1 strain having antimicrobial activity against microorganisms causing vaginosis and uses thereof | |
KR101982759B1 (en) | NOVEL STRAIN OF Bacillus subtilis AND COMPOSITION FOR PREVENTING OR TREATING OF PHATHOGEN BACTERIUM COMPRISING THE SAME |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AS | Assignment |
Owner name: KOBIOLABS, INC., KOREA, REPUBLIC OF Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:KO, GWANG PYO;YOU, HYUN JU;KWON, BO MI;REEL/FRAME:043858/0690 Effective date: 20170816 |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: DOCKETED NEW CASE - READY FOR EXAMINATION |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: NON FINAL ACTION MAILED |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: RESPONSE TO NON-FINAL OFFICE ACTION ENTERED AND FORWARDED TO EXAMINER |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: NON FINAL ACTION MAILED |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: RESPONSE TO NON-FINAL OFFICE ACTION ENTERED AND FORWARDED TO EXAMINER |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: FINAL REJECTION MAILED |
|
STCB | Information on status: application discontinuation |
Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION |