CN116036075B - A biological preparation for porcine actinobacillus pleuropneumoniae disease - Google Patents
A biological preparation for porcine actinobacillus pleuropneumoniae disease Download PDFInfo
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- CN116036075B CN116036075B CN202211341234.3A CN202211341234A CN116036075B CN 116036075 B CN116036075 B CN 116036075B CN 202211341234 A CN202211341234 A CN 202211341234A CN 116036075 B CN116036075 B CN 116036075B
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- actinobacillus pleuropneumoniae
- inula
- flowers
- pleuropneumoniae
- major
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- 241000606748 Actinobacillus pleuropneumoniae Species 0.000 title claims abstract description 44
- 201000010099 disease Diseases 0.000 title description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 title description 3
- 238000002360 preparation method Methods 0.000 title description 3
- 241000132446 Inula Species 0.000 claims abstract description 38
- 210000002919 epithelial cell Anatomy 0.000 claims abstract description 6
- 239000003814 drug Substances 0.000 claims description 7
- 229940079593 drug Drugs 0.000 claims description 6
- 241000533042 Inula grandiflora Species 0.000 claims 2
- 241000282887 Suidae Species 0.000 abstract description 6
- 210000004072 lung Anatomy 0.000 abstract description 5
- 239000003124 biologic agent Substances 0.000 abstract description 3
- 230000000844 anti-bacterial effect Effects 0.000 description 12
- 230000001580 bacterial effect Effects 0.000 description 7
- 238000012258 culturing Methods 0.000 description 7
- 230000002401 inhibitory effect Effects 0.000 description 7
- 230000005764 inhibitory process Effects 0.000 description 7
- 239000003242 anti bacterial agent Substances 0.000 description 5
- 229940088710 antibiotic agent Drugs 0.000 description 5
- 239000003899 bactericide agent Substances 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 210000004027 cell Anatomy 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000008868 Flower Essence Substances 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 206010035664 Pneumonia Diseases 0.000 description 3
- 238000004043 dyeing Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 241000606750 Actinobacillus Species 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 206010059866 Drug resistance Diseases 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 229920001202 Inulin Polymers 0.000 description 2
- 241000204031 Mycoplasma Species 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 230000010065 bacterial adhesion Effects 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 201000006509 pleuropneumonia Diseases 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 241000186046 Actinomyces Species 0.000 description 1
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 208000000059 Dyspnea Diseases 0.000 description 1
- 206010013975 Dyspnoeas Diseases 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
- 239000004793 Polystyrene Substances 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 229920004890 Triton X-100 Polymers 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 230000003266 anti-allergic effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000003235 crystal violet staining Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 239000000419 plant extract Substances 0.000 description 1
- 208000008423 pleurisy Diseases 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 208000023504 respiratory system disease Diseases 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/365—Lactones
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/70—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in livestock or poultry
Landscapes
- Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Epidemiology (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention provides a biological agent for actinobacillus pleuropneumoniae, belonging to the technical field of actinobacillus pleuropneumoniae treatment. The inula flowers can effectively inhibit the actinobacillus pleuropneumoniae, and can break the biological envelope of the actinobacillus pleuropneumoniae and inhibit the adhesion of the actinobacillus pleuropneumoniae to the lung epithelial cells of the pigs. Thus, inula flowers can be made into biological agents for the treatment of actinobacillus pleuropneumoniae.
Description
The application is a divisional application, the application name of the original application is that the inula major flowers are applied to the preparation of medicines for treating porcine actinobacillus pleuropneumoniae, the application date of the original application is 2020-11-06, and the application number of the original application is CN202011227605.6.
Technical Field
The invention relates to the technical field of treatment of actinobacillus pleuropneumoniae, in particular to a biological agent for actinobacillus pleuropneumoniae.
Background
With the wide clinical application of antibiotics, more and more pathogenic bacteria have developed resistance to antibiotics. The drug resistance of the germ causes serious loss to livestock and poultry cultivation. How to effectively combat bacterial infections has become a common concern worldwide, and alternative strategies to antibiotics have also attracted widespread human attention.
Actinobacillus pleuropneumoniae is also called as porcine infectious pleuropneumonia, and is one of respiratory diseases of a hazard-intensive pig farm caused by actinobacillus pleuropneumoniae, and is mainly characterized by pleurisy and pneumonia symptoms and lesions. This disease is often an acute illness and thus rapidly causes dyspnea in infected pigs and even rapid death of pigs, thus causing a great economic loss to the pig industry. At present, antibiotics are adopted for treatment aiming at the actinobacillus pleurodesis of pigs, and bacteria are easy to generate drug resistance, so that the selection of drugs capable of replacing the antibiotics is particularly important.
The inula flowers extract is a natural plant extract extracted from inula flowers. At present, the study on inula major flowers is relatively small, and the discovered biological functions mainly comprise anti-inflammatory, antiallergic, anti-tumor and the like. However, no report has been made about the inhibition of actinobacillus pleuropneumoniae by inula major.
Disclosure of Invention
The invention aims to provide application of inula flower essence in preparing medicines for treating porcine actinobacillus pleuropneumoniae.
In order to achieve the above purpose, the present invention provides the following technical solutions:
The invention provides application of inula major flowers in inhibiting actinobacillus pleuropneumoniae.
Preferably, the minimum inhibitory concentration of inula major flowers on actinobacillus pleuropneumoniae is 62.5ug/ml.
Preferably, the minimum bactericidal concentration of inula major flowers on actinobacillus pleuropneumoniae is 125ug/ml.
In addition, the invention provides application of inula flower essence in preparing a bactericide for inhibiting actinobacillus pleuropneumoniae.
In addition, the invention provides a bactericide for inhibiting actinobacillus pleuropneumoniae, and the core active ingredient of the bactericide is inula megaterium.
In addition, the invention provides application of inula major flowers in preparing bactericides for breaking biological membranes of actinobacillus pleuropneumoniae.
In addition, the invention provides application of inula major flowers in preparing bactericides for reducing bacterial adhesion of actinobacillus pleuropneumoniae.
In addition, the invention provides application of inula flower essence in preparing medicines for treating porcine actinobacillus pleuropneumoniae.
The beneficial effects of the invention are as follows:
Firstly, the invention discovers that the inula major flowers can effectively inhibit actinobacillus pleuropneumoniae;
Meanwhile, the invention discovers that the inula megaloba extract can break the biological envelope of actinobacillus pleuropneumoniae for the first time;
In addition, the invention discovers that the inula major flowers can inhibit the adhesion of actinobacillus pleuropneumoniae to pig lung epithelial cells for the first time. Thus, inula flowers can be used for preparing medicines for treating actinobacillus pleuropneumoniae.
Drawings
FIG. 1A diagram of the results of crystal violet staining of the biological envelope of Actinobacillus pleuropneumoniae
FIG. 2 graph of absorbance results of actinobacillus pleuropneumoniae biofilm
FIG. 3 is a graph showing the results of the adhesion inhibition rate of Actinobacillus pleuropneumoniae
Detailed Description
In order to clearly illustrate the technical characteristics of the scheme, the scheme is explained below through a specific embodiment.
Example 1
Minimum inhibitory concentration MIC and minimum bactericidal concentration MBC detection of inula major
Minimum inhibitory concentration detection
(1) The experimental groups were 500ug/ml,250ug/ml,125ug/ml,62.5ug/ml,31.25ug/ml,15.60ug/ml inula brinolysin and the negative control group ddH20;
(2) The number of actinobacillus pleuropneumoniae is regulated to 2X 10 5 CFU/ml, inoculated into a 96-well plate, and inula flowers (3 times repeated for each group of concentration) are added according to experimental groups, and the 96-well plate is placed into an incubator for culturing for 24 hours;
(3) The holes without turbidity are the minimum antibacterial concentration of the inula major flowers;
Minimum bactericidal concentration detection
(1) Continuously culturing bacteria for 24 hours, sucking bacterial liquid from a hole without turbidity, uniformly coating the bacterial liquid on an agar plate, putting the agar plate into a 37 ℃ incubator, culturing for 36 hours, and observing the result;
(2) The concentration of inula major flowers in wells with bacterial colony numbers not exceeding 5 was the minimum bactericidal concentration.
Experimental results show that the minimum inhibitory concentration of the inula megaterium is 62.5ug/ml, and the minimum bactericidal concentration is 125ug/ml.
Example 2
Sterilization effect detection for inula flowers
(1) Culturing actinobacillus pleuropneumoniae in a liquid culture medium until the OD600 is 1;
(2) The cells were grouped into inula flowers (125 ug/ml), positive control group (100 ug/ml penicillin), negative control group ddH20;
(3) Placing the strain into a shaking table at 37 ℃ for culturing, and detecting OD600 values of bacterial solutions at 2h,4h,6h,12h and 24h respectively.
From the table, it can be seen that inula megaflower has remarkable bactericidal effect on actinobacillus pleuropneumoniae, and the difference has statistical significance.
Example 3
Inhibition effect of inula major flower element on actinobacillus pleuropneumoniae biofilm
(1) The actinobacillus pleuropneumoniae in log phase is added into a 96-well polystyrene microplate;
(2) The experimental group is 125ug/ml inula flowers, and the control group is ddH2O;
(3) Treating actinobacillus pleuropneumoniae according to experimental groups, placing a miniature plate culture plate in an incubator for culturing for 24 hours, removing a culture medium, and adding sterile PBS for cleaning;
(4) Fixing with 70% methanol for 30min, removing fixing solution, and adding crystal violet for dyeing;
(5) After dyeing for 5min, removing the dyeing liquid, cleaning with water, and photographing;
(6) 70% ethanol was added to dissolve the biofilm and absorbance was measured at OD 570.
The experimental results are shown in fig. 1 and 2, and it can be seen from the figures that the inula flowers can significantly inhibit the biofilm of actinomyces pleuropneumoniae, and the difference has statistical significance (the control group is 2.117+/-0.156, the inula flowers are 0.113+/-0.055, and the P < 0.0001).
Example 4
Inhibition of actinobacillus pleuropneumoniae bacterial adhesion by inula megaterium
(1) Pig lung epithelial cells SJPL were inoculated in 24 well plates, 62.5ug/ml inulins, 125ug/ml inulins and blank (3 replicates per treatment setup) were added;
(2) Adding actinobacillus pleuropneumoniae of 1×10 8 CFU/ml, and culturing in a cell incubator for 6 hr;
(3) Removing the supernatant, gently washing the wells 3 times with PBS, and washing away non-adherent cells;
(4) 200ul of 0.1% Triton-X100 is added into each hole, after being mixed evenly by light shaking, the mixture is left at room temperature for 15min to fully lyse cells, and the liquid in the holes is collected;
(5) Performing plate colony counting after gradient dilution by normal saline;
(6) Adhesion inhibition (%) = (number of bacterial colonies of blank group-number of bacterial colonies of experimental group)/number of bacterial colonies of blank group×100%.
As shown in FIG. 3, it can be seen from the graph that the inhibition effect of 62.5ug/ml inula major on the adhesion of Actinobacillus pleuropneumoniae to pig lung epithelial cells SJPI is 40.67+ -1.528, and the adhesion inhibition effect of 125ug/ml inula major is 78.33+ -2.517. The results show that the inula flowers can obviously inhibit the adhesion effect of the actinobacillus pleuropneumoniae on the lung epithelial cells of the pigs, thereby reducing the probability of infection of the pigs with the actinobacillus pleuropneumoniae.
The technical features of the present invention that are not described in the present invention may be implemented by or using the prior art, and are not described in detail herein, but the above description is not intended to limit the present invention, and the present invention is not limited to the above examples, but is also intended to be within the scope of the present invention by those skilled in the art.
Claims (1)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211341234.3A CN116036075B (en) | 2020-11-06 | 2020-11-06 | A biological preparation for porcine actinobacillus pleuropneumoniae disease |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211341234.3A CN116036075B (en) | 2020-11-06 | 2020-11-06 | A biological preparation for porcine actinobacillus pleuropneumoniae disease |
| CN202011227605.6A CN112336715B (en) | 2020-11-06 | 2020-11-06 | Application of inula britannica extract in preparation of medicine for treating swine pleuropneumonia actinobacillus |
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| Application Number | Title | Priority Date | Filing Date |
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| CN202011227605.6A Division CN112336715B (en) | 2020-11-06 | 2020-11-06 | Application of inula britannica extract in preparation of medicine for treating swine pleuropneumonia actinobacillus |
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| CN116036075A CN116036075A (en) | 2023-05-02 |
| CN116036075B true CN116036075B (en) | 2025-02-25 |
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| CN202211341234.3A Active CN116036075B (en) | 2020-11-06 | 2020-11-06 | A biological preparation for porcine actinobacillus pleuropneumoniae disease |
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| CN112336715B (en) * | 2020-11-06 | 2022-11-04 | 青岛嘉智生物技术有限公司 | Application of inula britannica extract in preparation of medicine for treating swine pleuropneumonia actinobacillus |
| CN116687914B (en) * | 2023-08-07 | 2024-01-16 | 山东师范大学 | A drug to treat non-alcoholic fatty liver disease |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101518266A (en) * | 2008-02-27 | 2009-09-02 | 河北省农林科学院植物保护研究所 | Preparation method and application of inula japonica extract and combination thereof |
| CN112336715A (en) * | 2020-11-06 | 2021-02-09 | 青岛嘉智生物技术有限公司 | Application of inula britannica extract in preparation of medicine for treating swine pleuropneumonia actinobacillus |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5332572A (en) * | 1988-11-10 | 1994-07-26 | Iowa State University Research Foundation | Method for protection of swine against pleuropneumonia |
| KR100693077B1 (en) * | 2005-11-03 | 2007-03-12 | 채찬희 | Biological composition for the prevention and treatment of swine complex respiratory diseases |
| CN100396295C (en) * | 2006-04-30 | 2008-06-25 | 陈光华 | Medicine for treating virus hepatitis |
| CN102727486B (en) * | 2012-06-21 | 2013-12-25 | 中国人民解放军第二军医大学 | Application of Inula lineariifolia lactone A in preparation of medicine for treating myocarditis |
| CN104604861A (en) * | 2015-01-08 | 2015-05-13 | 馥稷生物科技发展（上海）有限公司 | Plant-source synergistic compound bactericide containing resveratrol and preparation method thereof |
| CN107625853A (en) * | 2017-09-08 | 2018-01-26 | 合肥申仁养殖有限公司 | Treat Chinese medicine composition of pig acute festering type gastroenteritis and preparation method thereof |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101518266A (en) * | 2008-02-27 | 2009-09-02 | 河北省农林科学院植物保护研究所 | Preparation method and application of inula japonica extract and combination thereof |
| CN112336715A (en) * | 2020-11-06 | 2021-02-09 | 青岛嘉智生物技术有限公司 | Application of inula britannica extract in preparation of medicine for treating swine pleuropneumonia actinobacillus |
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| CN116036075A (en) | 2023-05-02 |
| CN112336715A (en) | 2021-02-09 |
| CN112336715B (en) | 2022-11-04 |
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