CN114574396A - 一种益生菌组合物及其应用 - Google Patents
一种益生菌组合物及其应用 Download PDFInfo
- Publication number
- CN114574396A CN114574396A CN202210268752.0A CN202210268752A CN114574396A CN 114574396 A CN114574396 A CN 114574396A CN 202210268752 A CN202210268752 A CN 202210268752A CN 114574396 A CN114574396 A CN 114574396A
- Authority
- CN
- China
- Prior art keywords
- probiotic composition
- bee
- bifidobacterium
- lactobacillus
- staphylococcus aureus
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000006041 probiotic Substances 0.000 title claims abstract description 33
- 235000018291 probiotics Nutrition 0.000 title claims abstract description 33
- 230000000529 probiotic effect Effects 0.000 title claims abstract description 30
- 239000000203 mixture Substances 0.000 title claims abstract description 29
- 241000186000 Bifidobacterium Species 0.000 claims abstract description 9
- 241000186660 Lactobacillus Species 0.000 claims abstract description 9
- 229940039696 lactobacillus Drugs 0.000 claims abstract description 9
- 244000144987 brood Species 0.000 claims abstract description 8
- 241000186610 Lactobacillus sp. Species 0.000 claims abstract description 3
- 241000194032 Enterococcus faecalis Species 0.000 claims description 11
- 229940032049 enterococcus faecalis Drugs 0.000 claims description 11
- 238000002360 preparation method Methods 0.000 claims description 8
- 230000001580 bacterial effect Effects 0.000 claims description 6
- 241000191940 Staphylococcus Species 0.000 claims description 2
- 239000002778 food additive Substances 0.000 claims 1
- 241000191967 Staphylococcus aureus Species 0.000 abstract description 16
- 241000257303 Hymenoptera Species 0.000 abstract description 10
- 241001464948 Coprococcus Species 0.000 abstract description 9
- 239000002068 microbial inoculum Substances 0.000 abstract description 8
- 238000000338 in vitro Methods 0.000 abstract description 7
- 240000004355 Borago officinalis Species 0.000 abstract description 6
- 235000007689 Borago officinalis Nutrition 0.000 abstract description 6
- 239000000654 additive Substances 0.000 abstract description 6
- 230000000996 additive effect Effects 0.000 abstract description 6
- 238000009776 industrial production Methods 0.000 abstract description 2
- 208000024891 symptom Diseases 0.000 abstract description 2
- 230000000844 anti-bacterial effect Effects 0.000 abstract 1
- 241000256844 Apis mellifera Species 0.000 description 14
- 239000006228 supernatant Substances 0.000 description 13
- 238000002474 experimental method Methods 0.000 description 11
- 239000001963 growth medium Substances 0.000 description 11
- 201000010099 disease Diseases 0.000 description 9
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 9
- 239000000047 product Substances 0.000 description 9
- 241000894006 Bacteria Species 0.000 description 7
- 239000003242 anti bacterial agent Substances 0.000 description 7
- 229940088710 antibiotic agent Drugs 0.000 description 7
- 230000003385 bacteriostatic effect Effects 0.000 description 6
- 230000005764 inhibitory process Effects 0.000 description 6
- 239000000843 powder Substances 0.000 description 6
- 244000052616 bacterial pathogen Species 0.000 description 5
- 230000002401 inhibitory effect Effects 0.000 description 5
- 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 4
- 238000012258 culturing Methods 0.000 description 4
- 210000001035 gastrointestinal tract Anatomy 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 229920001817 Agar Polymers 0.000 description 3
- 241000282414 Homo sapiens Species 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- 239000003795 chemical substances by application Substances 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
- 238000011161 development Methods 0.000 description 3
- 230000000366 juvenile effect Effects 0.000 description 3
- 238000009630 liquid culture Methods 0.000 description 3
- LWIHDJKSTIGBAC-UHFFFAOYSA-K tripotassium phosphate Chemical compound [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 3
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- 241001478240 Coccus Species 0.000 description 2
- 241000238631 Hexapoda Species 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 241001468189 Melissococcus Species 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 208000024780 Urticaria Diseases 0.000 description 2
- 230000003213 activating effect Effects 0.000 description 2
- 230000010138 bee pollination Effects 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 239000002552 dosage form Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 229960003276 erythromycin Drugs 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 235000012907 honey Nutrition 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 238000007790 scraping Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- 206010067484 Adverse reaction Diseases 0.000 description 1
- 241000194103 Bacillus pumilus Species 0.000 description 1
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 208000019359 Enterococcus faecalis infection Diseases 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 206010061598 Immunodeficiency Diseases 0.000 description 1
- 208000029462 Immunodeficiency disease Diseases 0.000 description 1
- 241001468188 Melissococcus plutonius Species 0.000 description 1
- 241000193418 Paenibacillus larvae Species 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 108010034396 Streptogramins Proteins 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000012272 crop production Methods 0.000 description 1
- 238000012864 cross contamination Methods 0.000 description 1
- YXVFQADLFFNVDS-UHFFFAOYSA-N diammonium citrate Chemical compound [NH4+].[NH4+].[O-]C(=O)CC(O)(C(=O)O)CC([O-])=O YXVFQADLFFNVDS-UHFFFAOYSA-N 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
- 235000019797 dipotassium phosphate Nutrition 0.000 description 1
- 231100000676 disease causative agent Toxicity 0.000 description 1
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000007813 immunodeficiency Effects 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 244000000053 intestinal parasite Species 0.000 description 1
- 230000001418 larval effect Effects 0.000 description 1
- 229940041028 lincosamides Drugs 0.000 description 1
- 239000003120 macrolide antibiotic agent Substances 0.000 description 1
- 229940041033 macrolides Drugs 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 229940099596 manganese sulfate Drugs 0.000 description 1
- 235000007079 manganese sulphate Nutrition 0.000 description 1
- 239000011702 manganese sulphate Substances 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 235000013923 monosodium glutamate Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 231100000719 pollutant Toxicity 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 229910000160 potassium phosphate Inorganic materials 0.000 description 1
- 235000011009 potassium phosphates Nutrition 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 239000003223 protective agent Substances 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 229940073490 sodium glutamate Drugs 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 229940041030 streptogramins Drugs 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000001018 virulence Effects 0.000 description 1
- 239000012138 yeast extract 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
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/90—Feeding-stuffs specially adapted for particular animals for insects, e.g. bees or silkworms
-
- 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
- A23V2400/51—Bifidobacterium
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Polymers & Plastics (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Virology (AREA)
- Animal Husbandry (AREA)
- Biomedical Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biochemistry (AREA)
- Food Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Physiology (AREA)
- Molecular Biology (AREA)
- Tropical Medicine & Parasitology (AREA)
- General Engineering & Computer Science (AREA)
- Insects & Arthropods (AREA)
- Birds (AREA)
- Agronomy & Crop Science (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- Environmental Sciences (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
本发明公开了一种益生菌组合物及其应用，所述益生菌组合物由乳杆菌（Lactobacillus sp.）W8173和双歧杆菌（Bifidobacterium sp.）W8118组成；本发明益生菌组合物能改善蜜蜂欧洲幼虫腐臭病症状，在体外抑菌实验中，能够抑制蜂房蜜蜂球菌、金黄色葡萄球菌、粪球菌的生长，本发明益生菌组合物环保，无污染，不会影响蜂产品品质，可以作为菌剂、饲料或蜂粮添加剂，适用于工业化生产和市场推广应用。
Description
技术领域
本发明属于微生物技术领域，具体涉及一种益生菌组合物及其应用。
背景技术
蜜蜂是重要的经济昆虫，在构建与维持自然生态系统和生态农业、供给人类高级营养和保健功能产品的过程中扮演重要角色。在人类利用的粮食作物中，有76%依靠蜜蜂传花授粉。蜜蜂授粉产生的经济效益比蜂蜜产值还要高10至15倍。2019年5月6日，联合国生物多样性和生态系统服务政府间科学政策平台（IPBES）发布的权威报告显示，因为传粉动物的消失，全球农作物生产每年面临着约2350-5770亿美元的损失。但随着生态环境失衡的加剧，蜜蜂遭受病虫害、农药的威胁也逐渐加剧。其中蜜蜂细菌性疾病作为影响蜜蜂安全的主要危害之一，严重影响蜜蜂的生长发育，具有危害严重、传播迅速、持续时间长等特点，同时对蜂产品安全构成严重威胁。蜜蜂处于许多已知的病原体的威胁，其中为人所熟知的有革兰氏阳性菌蜂房蜜蜂球菌和幼虫芽孢杆菌引起的欧洲幼虫腐臭病和美洲幼虫腐臭病，主要针对蜜蜂幼虫并造成其大规模死亡。通常带有细菌病原体的成年蜜蜂，在采集花蜜及花粉时容易造成种群之间的交叉感染。革兰氏阳性菌蜂房蜜蜂球菌（Melissococcus pluton）被认为是欧幼病的诱因。蜂房蜜蜂球菌为兼性厌氧菌，生长需要的钠、钾比应等于或低于1，需要向培养基中添加磷酸钾来分离和维持该菌的生长。一些次级入侵者，如短小芽孢杆菌、粪肠球菌等也被认为是欧幼病的致病因子，但这种说法仍然存在争议，有学者认为次级入侵者不会增加欧幼病的致死率，可能仅仅是因为幼虫的免疫缺陷或死亡使得次级入侵者在幼虫体内定值。
金黄色葡萄球菌（Staphylococcus aureus）作为致病菌为人们而熟知，同时也是蜂粮的主要污染物。金黄色葡萄球菌的传统抑制方法仍以抗生素为主，临床上常使用大环内酯类(MLSB类)即(如红霉素)、林可酰胺类(如克林霉素)、B类链阳霉素类抗菌药物治疗金黄色葡萄球菌引起的感染。克林霉素因其半衰期长、不良反应少被广泛使用。然而，随着广谱抗生素的大量使用，多重耐药菌的发生呈上升趋势。金黄色葡萄球菌对克林霉素的耐药率逐年上升，仅次于青霉素和红霉素。同时抗生素的添加也降低了蜂蜜的质量，从而造成经济损失。
粪肠球菌（Enterococcus faecalis）属于动物肠道内的正常菌群，一般认为粪肠球菌对动物或人的机体无害，是一种共生菌。近年研究已证实，部分粪肠球菌在长期进化中出现毒力基因，可引起广泛的感染。而且由于抗生素的长期和大量不规范使用，粪肠球菌获得性耐药性不断上升，治疗粪肠球菌感染日益困难。
蜂场在治疗时因没有有效的治疗手段，滥用抗生素，不仅导致病源菌产生抗药性，而且会破坏蜜蜂肠道微生态平衡，使疾病进一步恶化；同时抗生素的过度施用，会导致其在蜂产品中的残留，影响产品品质，带来食品安全性威胁。当面临众多的肠道寄生虫和病原菌威胁时，肠道微生物能够通过产生抗菌物质来抑制肠道中病原菌微生物的生长。因此选用蜜蜂肠道中的益生菌作为蜂粮添加剂，是一种高效绿色的防治疾病的手段。
目前未见任何关于益生菌组合物用于欧洲幼虫腐臭病防治及蜂粮添加物的相关报道。
发明内容
针对现有技术存在的问题，本发明提供了一种益生菌组合物，所述益生菌组合物由乳杆菌（Lactobacillus sp.）W8173和双歧杆菌（Bifidobacterium sp.）W8118组成，乳杆菌W8173保藏编号为CGMCC NO.21787（202110510861.4）和双歧杆菌W8118的保藏编号为CGMCC NO.21787（202110509842.X），这些菌种均在其他专利文献中公开发表过。
本发明另一目的是将上述益生菌组合物应用在制备蜂粮添加剂中，防止金黄色葡萄球菌（Staphylococcus aureus）、粪肠球菌（Enterococcus faecalis）的污染。
本发明还可以将益生菌组合物应用在制备防治欧洲幼虫腐臭病的菌剂或饲料中，即以益生菌组合物或其发酵产物为活性成分，在制备防治欧洲幼虫腐臭病菌剂或饲料中的应用；
本发明防治欧洲幼虫腐臭病致病菌菌剂或饲料的成分（或有效成分）或者抑制金黄色葡萄球菌、粪肠球菌的添加剂为益生菌组合物或其发酵产物，还可以加入一种或多种防治菌剂或饲料上可接受的辅料，制成菌剂、饲料或添加剂适用的剂型，按常规添加量添加使用，有效活菌数不低于1011 cfu/g。
本发明的有益效果在于：
本发明旨在为蜜蜂的疾病防治和新型益生菌的市场化开发和产业化应用奠定基础；本发明提出的益生菌组合物，具有抑制蜂房蜜蜂球菌和金黄色葡萄球菌或粪球菌的作用，在治疗欧洲幼虫腐臭病和蜂粮的储备中有巨大潜力；
本发明提供的益生菌组合物，能改善蜜蜂欧洲幼虫腐臭病症状，在体外抑菌圈实验中，可以直接抑制蜂房蜜蜂球菌；同时在抑菌圈实验中发现益生菌组合物对金黄色葡萄球菌具有较好的抑制效果；且本发明益生菌组合制剂或饲料环保、无污染，不会影响蜂产品品质，适用于工业化生产和市场推广应用。
附图说明
图1为益生菌组合物上清液对蜂房蜜蜂球菌的体外抑菌实验结果；
图2为益生菌组合物上清液对金黄色葡萄球菌的体外抑菌实验结果；
图3为益生菌组合物上清液对粪球菌的体外抑菌实验结果。
具体实施方式
下面通过附图和实施例对本发明进一步说明，但本发明保护范围不局限于所述内容，本实施例中方法如无特殊说明的均按常规方法操作，所用试剂如无特殊说明均为常规市售试剂或按常规方法配置的试剂，培养基均为市售产品，按使用说明使用；
其中乳杆菌W8173所用培养基为MRS培养基：蛋白胨10g、牛肉膏10g、酵母提取物5g、磷酸氢二钾2g、柠檬酸二铵2g、乙酸钠5g、葡萄糖20g、吐温-80 1mL、硫酸镁0.5g、硫酸锰0.25g、琼脂粉15g，用蒸馏水定容至1L，将pH调至6.2-6.4；双歧杆菌W8118所用培养基为TPY培养基；
实施例1：冻干菌剂的制备
（1）将乳杆菌W8173用MRS培养基活化，双歧杆菌W8118用TPY培养基活化，接种量为3%，将活化好的菌种，按照4%接种量分别转接到MRS液体培养基和TPY液体培养基中，在37℃培养25h，离心(5000 g、15 min)收获菌体，备用；将上述菌体按照体积比为3:3:2加入保护剂（海藻糖1%、谷氨酸钠0.5%、脱脂乳1%、抗坏血酸0.25%、葡聚糖0.25%，g/100mL蒸馏水），混合均匀备用。在进行冷冻干燥之前，样品需在-80℃超低温冰箱预冻3h，随后采用三段式冷冻干燥，在512 Pa压力下冻干6 h，随后在256 Pa压力下冻干5 h，之后在103 Pa下冻干，直至获得粉末产品。将冻干的菌剂分别在4 ℃和25 ℃保存菌粉，每隔2个月测定每种菌粉的活菌数，平行3次实验。结果如下表所示；由表可知，冻干的菌剂在4 ℃和25 ℃保存10个月后，活菌数仍保持在较高的水平；在25 ℃保存下的稳定性较4 ℃保存下虽然有一定的差距，但是仍然保持在90%的存活水平；混合菌粉的有效菌数不低于2×1011 cfu/g。
实施例2：混合菌粉体外抑菌实验
（1）受试品混合菌粉无菌上清液的配置：将上述实验中的混合菌粉冻干菌剂，接种于BHI液体培养基中培养5天，将培养液通过低温高速离心机8000g离心10 min，取上层清液，最后将上层清液通过0.22 μm孔径滤膜过滤，得到益生菌组合物无菌上清液；
（2）致病菌蜂房蜜蜂球菌的培养及抑菌圈实验
在10mL离心管中加入5mL 1×PBS缓冲液，将在平板上处于最佳生长状态的蜂房蜜蜂球菌全部用接种环刮于离心管中，用涡旋振荡器振荡均匀，制备成菌悬液，取50μL菌悬液置于KSBHI平板上，涂布均匀；在涂布后的琼脂培养基上制备10mm的孔；取益生菌组合物无菌上清液100μL置于制备好的孔洞中；将平板置于5% CO2培养箱35℃培养3-4天；观察孔周围是否有抑制区域的形成。
结果见图1，从图中可以看出益生菌组合物的无菌上清液和对蜂房蜜蜂球菌具有较好的抑制作用，且比单一的乳杆菌W8173的抑菌效果好。
实施例3：益生菌组合物对金黄色葡萄球菌和粪球菌的体外抑菌实验
（1）无菌上清液的制备同实施例2步骤（1）；
（2）金黄色葡萄球菌及粪球菌的培养及抑菌圈实验
在10 mL离心管中加入5 mL 1×PBS缓冲液，将在平板上处于最佳生长状态的金黄色葡萄球菌及粪球菌全部用接种环刮于2个离心管中，用涡旋振荡器振荡均匀，制备成菌悬液，取50μL菌悬液置于BHI平板上，涂布均匀；在涂布后的琼脂培养基上制备10 mm的孔；取无菌上清液100 μL分别置于制备好的金黄色葡萄球菌及粪球菌的孔洞中；将平板置于5%CO2培养箱35℃培养3-4天；观察孔周围是否有抑制区域的形成；
由图2和图3结果可知，益生菌组合物的无菌上清液对金黄色葡萄球菌和粪球菌具有较好的抑制作用。
上述实验结果表明，本发明益生菌组合物能够有效抑制蜂房蜜蜂球菌、金黄色葡萄球菌、粪球菌的生长，为蜜蜂的疾病防治和新型益生菌的市场化开发和产业化应用奠定基础。
Claims (3)
1.一种益生菌组合物，其特征在于：由乳杆菌（Lactobacillus sp.）W8173和双歧杆菌（Bifidobacterium sp.）W8118组成。
2.权利要求1所述的益生菌组合物在制备蜂粮添加剂中的应用，其特征在于：益生菌组合物能抑制金黄色葡萄球菌（Staphylococcus aureus）和粪肠球菌（Enterococcus faecalis）。
3.权利要求1所述的益生菌组合物在制备治疗欧洲幼虫腐臭病的菌剂或饲料中的应用。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210268752.0A CN114574396B (zh) | 2022-03-18 | 2022-03-18 | 一种益生菌组合物及其应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210268752.0A CN114574396B (zh) | 2022-03-18 | 2022-03-18 | 一种益生菌组合物及其应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114574396A true CN114574396A (zh) | 2022-06-03 |
| CN114574396B CN114574396B (zh) | 2024-02-06 |
Family
ID=81777617
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210268752.0A Active CN114574396B (zh) | 2022-03-18 | 2022-03-18 | 一种益生菌组合物及其应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114574396B (zh) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103114051A (zh) * | 2012-11-15 | 2013-05-22 | 上海交大昂立股份有限公司 | 一种罗伊氏乳杆菌及其应用 |
| CN105176856A (zh) * | 2015-05-19 | 2015-12-23 | 浙江工商大学 | 一种清酒乳杆菌lz217筛选和培养方法及其应用 |
| US20210177963A1 (en) * | 2015-11-30 | 2021-06-17 | Joseph E. Kovarik | Method and System for Protecting Honey Bees from Pesticides |
| CN113215039A (zh) * | 2021-05-11 | 2021-08-06 | 中国农业大学 | 一株乳杆菌w8173及其应用 |
| CN113265350A (zh) * | 2021-05-11 | 2021-08-17 | 昆明理工大学 | 一种双歧杆菌w8118及其应用 |
| CN113265351A (zh) * | 2021-05-11 | 2021-08-17 | 昆明理工大学 | 一株乳杆菌w8172及其应用 |
-
2022
- 2022-03-18 CN CN202210268752.0A patent/CN114574396B/zh active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103114051A (zh) * | 2012-11-15 | 2013-05-22 | 上海交大昂立股份有限公司 | 一种罗伊氏乳杆菌及其应用 |
| CN105176856A (zh) * | 2015-05-19 | 2015-12-23 | 浙江工商大学 | 一种清酒乳杆菌lz217筛选和培养方法及其应用 |
| US20210177963A1 (en) * | 2015-11-30 | 2021-06-17 | Joseph E. Kovarik | Method and System for Protecting Honey Bees from Pesticides |
| CN113215039A (zh) * | 2021-05-11 | 2021-08-06 | 中国农业大学 | 一株乳杆菌w8173及其应用 |
| CN113265350A (zh) * | 2021-05-11 | 2021-08-17 | 昆明理工大学 | 一种双歧杆菌w8118及其应用 |
| CN113265351A (zh) * | 2021-05-11 | 2021-08-17 | 昆明理工大学 | 一株乳杆菌w8172及其应用 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114574396B (zh) | 2024-02-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| AU2008246387B2 (en) | Novel bacteria isolated from fresh honey or the honey producing tract of honey bees | |
| US20190247449A1 (en) | Compositions comprising a non-pathogenic bacteria and methods for protecting plant and animal hosts from fungal, bacterial and viral diseases | |
| NL2026421B1 (en) | Strain of clostridium perfringens phage, bacteriostatic agent including the phage, and preparation method and use thereof | |
| WO2009102143A2 (ko) | 김치유래 락토바실러스 플란타룸 bls41 및 그 용도 | |
| CN113040390B (zh) | 一株益生、耐盐约氏乳杆菌及其在畜禽水产养殖中防治病原菌的应用 | |
| WO2016046706A1 (en) | Probiotic fermented feed additives | |
| CN102356817A (zh) | 两株裂解性哈维弧菌噬菌体制剂的制备及其应用 | |
| Rajeswari et al. | Antimicrobial activities of cow dung extracts against human pathogens | |
| RU2437563C1 (ru) | Пробиотическая кормовая добавка | |
| WO2016027279A1 (en) | Novel bacterium of bacillus genus and uses thereof | |
| CN113215039A (zh) | 一株乳杆菌w8173及其应用 | |
| KR101818859B1 (ko) | 슈도모나스 아조토포만스 균주 kacc 92125p 및 이를 포함하는 조성물 | |
| KR101643235B1 (ko) | 황색포도상구균에 대해 숙주감수성을 나타내는 박테리오파지 시포비리데 패밀리 sap4 및 이를 함유하는 조성물 | |
| Subramanian et al. | Newparadigm in silkworm disease management using probiotic application of Streptomyces noursei | |
| JP2010136668A (ja) | 新規シュウドモナス属細菌 | |
| KR101596665B1 (ko) | 황색포도상구균에 대해 숙주감수성을 나타내는 박테리오파지 sa12 및 이를 함유하는 조성물 | |
| CN114574396B (zh) | 一种益生菌组合物及其应用 | |
| KR100371503B1 (ko) | 양식어류의 사료첨가제용 미생물 제제 및 그 제조방법 | |
| WO2023227648A1 (en) | Bacterial biocontrol agent | |
| El-Sheekh et al. | Antimicrobial activity of the cyanobacteria Anabaena wisconsinense and Oscillatoria curviceps against pathogens of fish in aquaculture | |
| Khalafalla et al. | Novel potential probiotics from gut microbiota of honeybees (Apis mellifera) in clover feeding season in Egypt. | |
| RU2292132C2 (ru) | Способ биологической борьбы с плесневением сена | |
| Ramesh et al. | Feasibility of shrimp gut probionts with anti-vibrio and anti-QS in penaeid culture | |
| CN113528461B (zh) | 分离的杀鲑气单胞菌噬菌体、其组合物及应用 | |
| RU2799554C1 (ru) | Новый пробиотический препарат на основе консорциума спорообразующих бактерий для аквакультуры и животных и способ его получения |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |