CN118045106A - Application of Burkholderia vietnamensis in preparation of biological preparation for inhibiting and/or killing Demodex - Google Patents
Application of Burkholderia vietnamensis in preparation of biological preparation for inhibiting and/or killing Demodex Download PDFInfo
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- CN118045106A CN118045106A CN202410185714.8A CN202410185714A CN118045106A CN 118045106 A CN118045106 A CN 118045106A CN 202410185714 A CN202410185714 A CN 202410185714A CN 118045106 A CN118045106 A CN 118045106A
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- burkholderia
- vietnaensis
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- 241001453380 Burkholderia Species 0.000 title claims abstract description 50
- 241001128004 Demodex Species 0.000 title claims abstract description 26
- 230000002147 killing effect Effects 0.000 title claims abstract description 21
- 230000002401 inhibitory effect Effects 0.000 title claims abstract description 15
- 238000002360 preparation method Methods 0.000 title claims abstract description 13
- 238000000855 fermentation Methods 0.000 claims abstract description 40
- 230000004151 fermentation Effects 0.000 claims abstract description 40
- 239000006228 supernatant Substances 0.000 claims abstract description 28
- 239000002609 medium Substances 0.000 claims description 18
- 239000003124 biologic agent Substances 0.000 claims description 13
- 239000007330 chocolate agar Substances 0.000 claims description 8
- 239000001963 growth medium Substances 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 7
- 238000000034 method Methods 0.000 claims description 7
- 239000007787 solid Substances 0.000 claims description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 4
- 239000001888 Peptone Substances 0.000 claims description 4
- 108010080698 Peptones Proteins 0.000 claims description 4
- 239000001110 calcium chloride Substances 0.000 claims description 4
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 4
- 229940041514 candida albicans extract Drugs 0.000 claims description 4
- 238000009630 liquid culture Methods 0.000 claims description 4
- 235000019319 peptone Nutrition 0.000 claims description 4
- 239000012138 yeast extract Substances 0.000 claims description 4
- 230000001954 sterilising effect Effects 0.000 claims description 3
- 238000000338 in vitro Methods 0.000 abstract description 4
- 241000238876 Acari Species 0.000 description 6
- 230000004083 survival effect Effects 0.000 description 5
- 239000003708 ampul Substances 0.000 description 4
- 230000000694 effects Effects 0.000 description 3
- 241000238631 Hexapoda Species 0.000 description 2
- 208000010217 blepharitis Diseases 0.000 description 2
- 208000002874 Acne Vulgaris Diseases 0.000 description 1
- 206010067484 Adverse reaction Diseases 0.000 description 1
- 239000005995 Aluminium silicate Substances 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 206010016936 Folliculitis Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 208000023715 Ocular surface disease Diseases 0.000 description 1
- 241001303601 Rosacea Species 0.000 description 1
- 206010000496 acne Diseases 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 235000012211 aluminium silicate Nutrition 0.000 description 1
- 235000019606 astringent taste Nutrition 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 210000000720 eyelash Anatomy 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- NLYAJNPCOHFWQQ-UHFFFAOYSA-N kaolin Chemical compound O.O.O=[Al]O[Si](=O)O[Si](=O)O[Al]=O NLYAJNPCOHFWQQ-UHFFFAOYSA-N 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 244000045947 parasite Species 0.000 description 1
- 230000007918 pathogenicity Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 201000004700 rosacea Diseases 0.000 description 1
- 238000011076 safety test Methods 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 229940111630 tea tree oil Drugs 0.000 description 1
- 239000010677 tea tree oil Substances 0.000 description 1
Classifications
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P33/00—Antiparasitic agents
- A61P33/14—Ectoparasiticides, e.g. scabicides
-
- 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
-
- 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
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Animal Behavior & Ethology (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Molecular Biology (AREA)
- Mycology (AREA)
- Epidemiology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention belongs to the technical field of mite-inhibiting and killing preparations, and relates to application of Vietnam burkholderia in preparing biological preparations for inhibiting and/or killing Demodex. The Vietnam burkholderia fermentation supernatant provided by the invention has high safety and has the function of killing Demodex in vitro.
Description
Technical Field
The invention belongs to the technical field of mite-inhibiting and killing preparations, and particularly relates to application of Burkholderia vietnamensis in preparation of a biological preparation for inhibiting and/or killing Demodex.
Background
Demodex mites are common parasites of humans and have a certain pathogenicity. Commonly caused diseases include rosacea, folliculitis, acne, blepharitis, and the like. Acarid blepharitis is a common ocular surface disease that can cause discomfort such as drop of eyelashes, red eyes, astringency of eyes, tearing, etc. At present, the main treatment methods are tea tree oil and antibiotic eye cream, but the effect is not ideal, and some patients have incomplete mite removal and are easy to relapse. At present, no biological agent for killing mites exists.
Disclosure of Invention
The invention aims to provide application of Vietnam burkholderia in preparing biological agents for inhibiting the growth of Demodex and/or killing Demodex. The invention discovers that the fermentation supernatant of Burkholderia vietnamensis has the function of killing Demodex in vitro.
The invention provides application of Vietnam burkholderia fermentation supernatant in preparing a biological agent for inhibiting the growth of Demodex and/or killing Demodex.
Preferably, the burkholderia vietnaensis is burkholderia vietnamensis SHBCCD 51593=atcc BAA-248=lmg 10929, purchased from the Shanghai collection biotechnology center.
The invention also provides a biological agent for inhibiting the growth of Demodex and/or killing Demodex, the biological agent comprising Vietnam Burkholderia fermentation supernatant.
Preferably, the burkholderia vietnaensis is burkholderia vietnamensis SHBCCD 51593=atcc BAA-248=lmg 10929, purchased from the Shanghai collection biotechnology center.
The invention also provides a preparation method of the Vietnam Burkholderia fermentation supernatant in the application of the technical scheme, which comprises the following steps:
Inoculating Burkholderia vietnaensis to a solid culture medium for subculture, inoculating to a liquid culture medium for fermentation culture after subculture, and centrifuging to obtain a Burkholderia vietnaensis fermentation supernatant.
Preferably, the solid medium comprises chocolate agar medium.
Preferably, the liquid medium comprises: 10g/L peptone, 1g/L yeast extract, 0.2g/L calcium chloride, and sterilizing water, and the pH value is 7.2-7.4.
Preferably, the temperature of the subculture is 35-37 ℃; the fermentation culture time is 24-72 hours; the times of subculture are 1-3 times.
Preferably, the temperature of the fermentation culture is 35-37 ℃, and the time of the fermentation culture is 24-72 hours; the rotation speed of the fermentation culture is 150-300 rpm.
Preferably, the centrifugal speed is 7000 to 9000rpm.
The invention provides application of Vietnam burkholderia fermentation supernatant in preparing a biological agent for inhibiting the growth of Demodex and/or killing Demodex. The invention discovers that the Vietnam Burkholderia fermentation supernatant has the function of killing Demodex in vitro for the first time, and the Vietnam Burkholderia fermentation supernatant has high safety and can kill Demodex.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions of the prior art, the drawings that are needed in the embodiments will be briefly described below, it being obvious that the drawings in the following description are only some embodiments of the present invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a graph showing the results of killing Demodex mites with the liquid medium and Vietnam Burkholderia fermentation supernatant provided by the invention;
fig. 2 is a diagram of safety test results provided by the invention.
Detailed Description
The invention provides application of Vietnam burkholderia fermentation supernatant in preparing a biological agent for inhibiting the growth of Demodex and/or killing Demodex. In the present invention, the burkholderia vietnaensis is burkholderia vietnaensis SHBCCD 51593=atcc BAA-248=lmg 10929, purchased from Shanghai collection biotechnology center. The invention discovers that the Vietnam Burkholderia fermentation supernatant has the effect of killing Demodex mites in vitro for the first time, and particularly shortens the survival time of mites.
The invention also provides a biological agent for inhibiting the growth of Demodex and/or killing Demodex, the biological agent comprising Vietnam Burkholderia fermentation supernatant. In the present invention, the burkholderia vietnaensis is burkholderia vietnaensis SHBCCD 51593=atcc BAA-248=lmg 10929, purchased from Shanghai collection biotechnology center.
The invention also provides a preparation method of the Vietnam Burkholderia fermentation supernatant in the application of the technical scheme, which comprises the following steps:
Inoculating Burkholderia vietnaensis to a solid culture medium for subculture, inoculating to a liquid culture medium for fermentation culture after subculture, and centrifuging to obtain a Burkholderia vietnaensis fermentation supernatant.
The invention inoculates Burkholderia vietnaensis into a solid culture medium for subculture. In the present invention, the solid medium preferably comprises a chocolate agar medium. The source of the chocolate agar medium is not particularly limited, and the chocolate agar medium is a conventional commercial chocolate agar medium, such as the chocolate agar medium purchased from Jiangmen Kaolin trade company.
After subculture, the strain is inoculated to a liquid culture medium for fermentation culture and centrifuged to obtain Vietnam Burkholderia fermentation supernatant. In the present invention, the liquid medium preferably includes: 10g/L peptone, 1g/L yeast extract, 0.2g/L calcium chloride, and sterilizing water, and the pH value is 7.2-7.4. In the present invention, the temperature of the subculture is preferably 35 to 37 ℃, more preferably 37 ℃; the fermentation culture time is preferably 24-72 hours, more preferably 48 hours; the number of subcultures is preferably 1 to 3, more preferably 2. In the present invention, the temperature of the fermentation culture is preferably 35 to 37 ℃, more preferably 37 ℃, and the time of the fermentation culture is preferably 24 to 72 hours, more preferably 48 hours; the rotation speed of the fermentation culture is preferably 150 to 300rpm, more preferably 200rpm. In the present invention, the centrifugal rotational speed is preferably 7000 to 9000rpm, more preferably 8000rpm.
For further explanation of the present invention, the application of the vietnam burkholderia of the present invention in the preparation of biological agents for inhibiting the growth of and/or killing demodex mites is described in detail below with reference to the accompanying drawings and examples, but they should not be construed as limiting the scope of the present invention.
Example 1
Burkholderia vietnaensis model strain: purchased from Shanghai collection biotechnology center, burkholderia vietnam SHBCCD 51593=atcc BAA-248=lmg 10929.
The Vietnam Burkholderia powder is thrown to the bottom of an ampoule in a biosafety cabinet, and the ampoule is knocked out after the top of the ampoule is sterilized by alcohol. 0.1ml of sterile water was added to the ampoule, gently flicked, mixed well, the solution was aspirated with a sterile pipette, and inoculated into chocolate agar medium (available from Jiangmen Kaili trade Co., ltd.). After 48h of culture in a 37 ℃ incubator, subculturing for 2 generations, placing the culture in a liquid medium (peptone 10g/L, yeast extract 1g/L, calcium chloride 0.2g/L, sterilized water for dissolution, pH 7.2) and culturing for 48h at 200rpm in a 37 ℃ incubator. Followed by centrifugation at 8000rpm to obtain a fermentation supernatant. The supernatant is preserved aseptically in a refrigerator at 4 ℃.
Mites were placed in the fermentation supernatant stock and their survival time was recorded by observation.
Judgment criteria for mite survival: the limb joints of the insects are kept still for 2min, and the body surfaces of the insects shrink, change color, deform, cause the contents to flow outwards, and the like.
In order to more accurately record the mite death time, after the first found mite death, it is confirmed again at the next observation. If the mites are still not obviously active in the second observation, determining that the mites die, and determining the time when the death characteristics appear for the first time as the death time; on the contrary, mite death cannot be determined and needs to be observed continuously.
Observation of mite-killing effect of Vietnam Burkholderia fermentation supernatant: mite survival times in liquid medium and in Vietnam Burkholderia fermentation supernatants were compared. Each group included 15 mites and the test results are shown in figure 1. The survival time in liquid medium (control) was about 58.+ -. 10.06 hours, and in Burkholderia vietnaesite fermentation supernatant (experimental) was about 49.4.+ -. 8.32 hours.
Example 2
The supernatant of Burkholderia vietnamensis is smeared on the skin surface of a mouse, and no obvious inflammation stimulus such as red swelling is seen for 48 hours. The material was drawn and CD3 stained, and no significant increase was seen in experimental group CD3 compared to control group. As shown in fig. 2. Conclusion: the result shows that the supernatant of the Burkholderia vietnaensis does not irritate the skin and causes adverse reaction, and has higher safety.
Although the foregoing embodiments have been described in some, but not all, embodiments of the invention, it should be understood that other embodiments may be devised in accordance with the present embodiments without departing from the spirit and scope of the invention.
Claims (10)
1. Application of Vietnam burkholderia fermentation supernatant in preparing biological preparation for inhibiting and/or killing Demodex.
2. The use according to claim 1, wherein the burkholderia vietnaensis is burkholderia vietnaensis SHBCCD 51593=atcc BAA-248=lmg 10929, available from the Shanghai collection biotechnology center.
3. A biological agent for inhibiting and/or killing demodex mites, characterized in that said biological agent comprises a fermentation supernatant of burkholderia vietnaesite.
4. A biological agent according to claim 3, wherein the burkholderia vietnam is burkholderia vietnam SHBCCD51593 = ATCC BAA-248 = LMG10929, available from the Shanghai collection biotechnology center.
5. The method for preparing the fermentation supernatant of Burkholderia vietnaensis for use according to claim 1, comprising the steps of:
Inoculating Burkholderia vietnaensis to a solid culture medium for subculture, inoculating to a liquid culture medium for fermentation culture after subculture, and centrifuging to obtain a Burkholderia vietnaensis fermentation supernatant.
6. The method of claim 5, wherein the solid medium comprises chocolate agar medium.
7. The method of claim 5, wherein the liquid medium comprises: 10g/L peptone, 1g/L yeast extract, 0.2g/L calcium chloride, and sterilizing water, and the pH value is 7.2-7.4.
8. The method according to claim 5, wherein the subculture temperature is 35 to 37 ℃; the fermentation culture time is 24-72 hours; the times of subculture are 1-3 times.
9. The method according to claim 5, wherein the fermentation culture is carried out at a temperature of 35 to 37 ℃ for 24 to 72 hours; the rotation speed of the fermentation culture is 150-300 rpm.
10. The method according to claim 5, wherein the centrifugal rotational speed is 7000 to 9000rpm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410185714.8A CN118045106A (en) | 2024-02-19 | 2024-02-19 | Application of Burkholderia vietnamensis in preparation of biological preparation for inhibiting and/or killing Demodex |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410185714.8A CN118045106A (en) | 2024-02-19 | 2024-02-19 | Application of Burkholderia vietnamensis in preparation of biological preparation for inhibiting and/or killing Demodex |
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| CN118045106A true CN118045106A (en) | 2024-05-17 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202410185714.8A Pending CN118045106A (en) | 2024-02-19 | 2024-02-19 | Application of Burkholderia vietnamensis in preparation of biological preparation for inhibiting and/or killing Demodex |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013032693A2 (en) * | 2011-08-27 | 2013-03-07 | Marrone Bio Innovations, Inc. | Isolated bacterial strain of the genus burkholderia and pesticidal metabolites therefrom-formulations and uses |
| WO2015034629A1 (en) * | 2013-09-07 | 2015-03-12 | Marrone Bio Innovations, Inc. | Methods and compositions for control of mite infestations using a newly discovered species of burkholderia |
| WO2018141063A1 (en) * | 2017-02-02 | 2018-08-09 | Mcmaster University | Bicarbonate as a potentiator for antimicrobial agents |
| WO2020198414A1 (en) * | 2019-03-26 | 2020-10-01 | University Of Florida Research Foundation | Compositions and methods relating to insecticides |
| CN116348589A (en) * | 2020-08-03 | 2023-06-27 | 巴斯夫欧洲公司 | Materials and methods for improving plant health |
-
2024
- 2024-02-19 CN CN202410185714.8A patent/CN118045106A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013032693A2 (en) * | 2011-08-27 | 2013-03-07 | Marrone Bio Innovations, Inc. | Isolated bacterial strain of the genus burkholderia and pesticidal metabolites therefrom-formulations and uses |
| WO2015034629A1 (en) * | 2013-09-07 | 2015-03-12 | Marrone Bio Innovations, Inc. | Methods and compositions for control of mite infestations using a newly discovered species of burkholderia |
| WO2018141063A1 (en) * | 2017-02-02 | 2018-08-09 | Mcmaster University | Bicarbonate as a potentiator for antimicrobial agents |
| WO2020198414A1 (en) * | 2019-03-26 | 2020-10-01 | University Of Florida Research Foundation | Compositions and methods relating to insecticides |
| CN116348589A (en) * | 2020-08-03 | 2023-06-27 | 巴斯夫欧洲公司 | Materials and methods for improving plant health |
Non-Patent Citations (3)
| Title |
|---|
| DULEI ZOU等: "Characteristics of bacterial community in eyelashes of patients with Demodex blepharitis", PARASITES & VECTORS, vol. 17, 14 February 2024 (2024-02-14), pages 10 - 11 * |
| PAOLA CESCUTTI等: "Structure of a novel exopolysaccharide produced by Burkholderia vietnamiensis, a cystic fibrosis opportunistic pathogen", CARBOHYDRATE POLYMERS, vol. 94, no. 1, 15 April 2013 (2013-04-15), pages 253 * |
| 顾林玲: "MBI生物农药Venerate获美国加州登记 提升特定作物产量达80%", 农药, vol. 58, no. 3, 10 March 2019 (2019-03-10), pages 203 * |
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