CN112516190A - Feed with antibacterial function and preparation method thereof - Google Patents
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- CN112516190A CN112516190A CN202011578631.3A CN202011578631A CN112516190A CN 112516190 A CN112516190 A CN 112516190A CN 202011578631 A CN202011578631 A CN 202011578631A CN 112516190 A CN112516190 A CN 112516190A
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- 230000000844 anti-bacterial effect Effects 0.000 title claims description 27
- 238000002360 preparation method Methods 0.000 title abstract description 9
- 239000000284 extract Substances 0.000 claims abstract description 59
- 230000003385 bacteriostatic effect Effects 0.000 claims abstract description 56
- 244000025254 Cannabis sativa Species 0.000 claims abstract description 52
- 239000000453 juniperus communis l. leaf oil Substances 0.000 claims abstract description 46
- SPFYMRJSYKOXGV-UHFFFAOYSA-N Baytril Chemical compound C1CN(CC)CCN1C(C(=C1)F)=CC2=C1C(=O)C(C(O)=O)=CN2C1CC1 SPFYMRJSYKOXGV-UHFFFAOYSA-N 0.000 claims abstract description 34
- 229960000740 enrofloxacin Drugs 0.000 claims abstract description 34
- 238000002156 mixing Methods 0.000 claims abstract description 33
- 238000002386 leaching Methods 0.000 claims abstract description 28
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 24
- 239000000843 powder Substances 0.000 claims abstract description 22
- 238000001035 drying Methods 0.000 claims abstract description 11
- 238000000227 grinding Methods 0.000 claims abstract description 11
- 201000000306 sarcoidosis Diseases 0.000 claims abstract description 11
- 244000162475 Juniperus rigida Species 0.000 claims abstract description 7
- 235000009069 Juniperus rigida Nutrition 0.000 claims abstract description 7
- 239000000203 mixture Substances 0.000 claims description 152
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 19
- 238000010438 heat treatment Methods 0.000 claims description 17
- 238000001816 cooling Methods 0.000 claims description 12
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- 229940079593 drug Drugs 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 10
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- 230000002401 inhibitory effect Effects 0.000 claims description 3
- 230000001580 bacterial effect Effects 0.000 abstract description 15
- 241000251468 Actinopterygii Species 0.000 description 17
- 230000005764 inhibitory process Effects 0.000 description 11
- 230000035945 sensitivity Effects 0.000 description 10
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- 241000894006 Bacteria Species 0.000 description 7
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- 230000009278 visceral effect Effects 0.000 description 7
- 241000607545 Aeromonas schubertii Species 0.000 description 6
- 241000721662 Juniperus Species 0.000 description 6
- 241001503642 Nocardia seriolae Species 0.000 description 6
- 238000009360 aquaculture Methods 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 240000005308 Juniperus chinensis Species 0.000 description 5
- 201000010099 disease Diseases 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
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- 230000000694 effects Effects 0.000 description 4
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- 206010020751 Hypersensitivity Diseases 0.000 description 3
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- 208000035143 Bacterial infection Diseases 0.000 description 2
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- 229940088710 antibiotic agent Drugs 0.000 description 2
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- 239000006161 blood agar Substances 0.000 description 1
- 241001233037 catfish Species 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 229940067866 dandelion extract Drugs 0.000 description 1
- 235000020691 dandelion extract Nutrition 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
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- 238000007598 dipping method Methods 0.000 description 1
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- 238000002649 immunization Methods 0.000 description 1
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Classifications
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- 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
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/116—Heterocyclic compounds
- A23K20/137—Heterocyclic compounds containing two hetero atoms, of which at least one is nitrogen
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- 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
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/13—Coniferophyta (gymnosperms)
- A61K36/14—Cupressaceae (Cypress family), e.g. juniper or cypress
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
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- 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
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- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/15—Preparation or pretreatment of starting material involving mechanical treatment, e.g. chopping up, cutting or grinding
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- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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Abstract
The invention provides a feed with a bacteriostatic function and a preparation method thereof, wherein the juniper extract is prepared by the following steps: drying Juniperus Rigida, grinding into powder, adding the powder into 10-20 times of 50-70% ethanol, leaching for 1-3 hr, repeatedly leaching for 3-5 times, mixing leaching solutions, and concentrating to obtain Juniperus Rigida Lindl extract. The feed with the bacteriostatic function effectively inhibits bacterial sarcoidosis by adding enrofloxacin, juniper extract and dancing grass extract.
Description
Technical Field
The invention belongs to the field of bacteriostasis, and particularly relates to a feed with a bacteriostasis function and a preparation method thereof.
Background
China is a big aquaculture country, the aquaculture density and the yield of aquatic products are high, but the fish bacterial diseases frequently occur due to the laggard aquaculture technology and facilities. At present, the drug prevention and treatment mainly based on antibiotics is still the most effective and most direct method in the culture production, and plays an important role in preventing and treating fish bacterial diseases. However, the long-term use of antibiotics in production practice can lead to the emergence of various drug-resistant strains in a short time.
The visceral sarcoidosis of fishes, commonly known as fish viscera-white spot disease |, in aquaculture is a general name for diseases which can cause the visceral tissues and organs of diseased fishes to have white or yellow punctate structural symptoms, and the punctate structures are similar to nodules through visual observation and are also known as the sarcoidosis |. In recent years, the reports on visceral sarcoidosis of fishes are more, infected hosts are wide, dozens of major cultured fishes such as snakehead, weever, catfish, large yellow croaker and the like are damaged, and huge economic loss is caused to aquaculture industry. Various diseases causing the fish visceral nodule exist, some diseases are chronic consumption diseases for a long time, the growth of a host is influenced, and the culture cost is increased; some of them can cause fulminant death and cause direct loss, and their pathogeny, infection host species and pathogenesis are very different.
The pathogeny of the visceral sarcoidosis of the fishes reported at present comprises bacteria, parasites, rickettsia-like bodies and the like, wherein the bacterial visceral sarcoidosis of the fishes is the most common and the harm to the aquaculture industry is the most serious. There is no effective means for inhibiting the visceral sarcoidosis of bacterial fish.
Disclosure of Invention
In view of the above, the invention provides a feed with a bacteriostatic function and a preparation method thereof, aiming at overcoming the defects in the prior art.
In order to achieve the purpose, the technical scheme of the invention is realized as follows:
an application of feed with bacteriostatic function in preparing medicines for inhibiting pathogenic bacteria of sarcoidosis.
Further, the feed comprises 0.01-1% of the bacteriostatic composition.
A bacteriostatic composition comprises the following components in a mass ratio of 1: 1-100: 100-1000 parts of enrofloxacin, juniper extract and dancing grass extract.
Further, the bacteriostatic composition comprises the components in a mass ratio of 1: 1-100: 100-1000 parts of enrofloxacin, juniper extract and dancing grass extract.
Preferably, the mass ratio of the enrofloxacin, the juniper extract and the dancing grass extract is 1: 1-50: 500-1000; the mass ratio of the enrofloxacin, the juniper extract and the dancing grass extract is 1: 35-50: 620-800.
Further, the juniper extract is prepared by the method comprising the following steps: drying Juniperus Rigida, grinding into powder, adding the powder into 10-20 times of 50-70% ethanol, leaching for 1-3 hr, repeatedly leaching for 3-5 times, mixing leaching solutions, and concentrating to obtain Juniperus Rigida Lindl extract.
Further, the dancing grass extract is prepared by the method comprising the following steps: drying herba Trifolii Pratentis, grinding into powder, adding the powder into 10-20 times of 60-70% ethanol, leaching for 1-3 hr, and concentrating the leaching solution to obtain the extract.
The preparation method of the antibacterial composition comprises the steps of heating the dancing grass extract in a water bath at 40-80 ℃ for 10-30 minutes, then slowly dripping the juniper extract into the dancing grass extract, uniformly mixing to obtain a mixture, cooling the mixture to room temperature, then adding enrofloxacin into the mixture, and uniformly mixing to obtain the antibacterial composition.
Further, the temperature of the water bath heating step is 50-65 ℃, and the time is 15-25 minutes.
A feed with bacteriostatic function comprises 0.01-1% of bacteriostatic composition; the antibacterial composition comprises the following components in a mass ratio of 1: 1-100: 100-1000 parts of enrofloxacin, juniper extract and dancing grass extract.
Compared with the prior art, the invention has the following advantages:
the feed with the bacteriostatic function effectively inhibits bacterial sarcoidosis by adding enrofloxacin, juniper extract and dancing grass extract.
Detailed Description
Unless defined otherwise, technical terms used in the following examples have the same meanings as commonly understood by one of ordinary skill in the art to which the present invention belongs. The test reagents used in the following examples, unless otherwise specified, are all conventional biochemical reagents; the experimental methods are conventional methods unless otherwise specified.
The mycobacteria (Mycobacterium), Nocardia seriolae (Nocardia seriolae) and Aeromonas schubertii (Aeromonas schubertii) described in the examples of the present invention were presented to the aquatic disease and immunization research laboratory of the Zhujiang aquatic research institute, the institute of Aquaculture, China.
The present invention will be described in detail with reference to examples.
Example 1 preparation of feedstuff having bacteriostatic action
Accurately weighing 500g of juniper chinensis, drying, grinding into powder, adding the powder into 10 times of 60% ethanol, leaching for 2 hours, repeatedly leaching for 5 times, mixing leaching solutions, and concentrating to obtain juniper chinensis extract.
Weighing 5000g of dancing grass accurately, drying, grinding into powder, adding the powder into 10 times of 70% ethanol, leaching for 3 hours, and concentrating the leaching solution to obtain the dancing grass extract.
Heating 750g of the dancing grass extract in a water bath at 60 ℃ for 20 minutes, slowly dripping 40g of the juniper extract into the dancing grass extract, uniformly mixing to obtain a mixture, cooling the mixture to room temperature, then adding 1g of enrofloxacin into the mixture, and uniformly mixing to obtain the antibacterial composition A.
Heating 750g of the dancing grass extract in a water bath at 45 ℃ for 20 minutes, slowly dripping 40g of the juniper extract into the dancing grass extract, uniformly mixing to obtain a mixture, cooling the mixture to room temperature, then adding 1g of enrofloxacin into the mixture, and uniformly mixing to obtain the antibacterial composition B.
Heating 750g of the dancing grass extract in a water bath at 90 ℃ for 20 minutes, slowly dripping 40g of the juniper extract into the dancing grass extract, uniformly mixing to obtain a mixture, cooling the mixture to room temperature, then adding 1g of enrofloxacin into the mixture, and uniformly mixing to obtain the antibacterial composition C.
Adding 40g of juniper extract into 750g of dancing grass extract, uniformly mixing, then adding 1g of enrofloxacin, and uniformly mixing to obtain the antibacterial composition D.
EXAMPLE 2 bacteriostatic Effect of the bacteriostatic composition on mycobacteria
1. Preparation of the culture Medium
BHI medium was purchased from BD, usa; 5% sheep blood agar plates, bacteria biochemical microassay tubes, etc. were purchased from Kyowa microorganism Co., Ltd.
2. Preparation of the bacterial suspension
And (3) test bacterium culture: activating the mycobacterium strains, streaking on a solid culture medium by using an inoculating needle, culturing for 24 hours at 32 ℃, picking single colonies, inoculating on a nutrient broth culture medium, and culturing for 24 hours with shaking. And (3) diluting the bacterial liquid to an appropriate concentration by adopting a plate counting method, wherein the concentration of the bacterial liquid is 1 multiplied by 105 CFU/mL.
3. Drug susceptibility testing
The drug sensitivity test adopts an agar diffusion method, bacterial suspension cultured to a logarithmic phase is diluted to the concentration of about 1 × 105CFU/mL, a sterile cotton swab is used for dipping bacterial liquid and uniformly smearing the bacterial liquid on a culture medium, after the bacterial liquid is uniformly smeared, holes are uniformly punched on the plates, 4 holes are punched on each plate, and the marks are made. Add 100. mu.l (1g/mL) of the composition to each well, then place it in an incubator at 32 ℃ for 24 h; and (4) after the flat plate is taken out, observing whether a bacteriostatic circle exists around the hole, and if so, measuring the diameter of the bacteriostatic circle by using a vernier caliper. Three replicates of each drug were run with a blank of sterile water. The result judgment standard is according to the standard introduced in the Chinese medicine pharmacology: the inhibition zone is more than or equal to 20mm, and the drug is extremely sensitive; the inhibition zone is 15-19mm, belonging to high sensitivity; the zone of inhibition is 10-14mm, belonging to the middle allergy; the inhibition zone is less than 10mm, and the low-sensitivity drug is low-sensitivity drug; has no bacteriostatic zone, and is insensitive (no bacteriostatic effect).
4. Secondary screening test
The extracts were diluted in a two-fold dilution with nutrient broth, the bacterial suspension was added to each tube to a bacterial concentration of 106CFU/mL, and composition a, composition B, composition C, composition E, no composition as blank and no bacteria as negative control, each repeated 3 times, in different tubes. Shaking in a shaking table at 32 deg.C (180r/min) for 24h, taking out, comparing with a control tube, and observing, wherein the diameter of the inhibition zone and the inhibition grade are shown in Table 1.
TABLE 1 zone diameter and grade of inhibition
| Item | Diameter of bacteriostatic circle (mm) | Grade of bacteriostasis |
| Composition A | 21.36±0.22 | +++ |
| Composition B | 17.84±0.26 | ++ |
| Composition C | 10.86±0.31 | + |
| Composition D | 13.28±0.36 | + |
| Blank control | — | — |
| Negative control | — | — |
In the above tables, "+ + + + +" indicates extreme sensitivity, "+" indicates medium sensitivity or low sensitivity, and "-" indicates no sensitivity (no bacteriostatic effect).
As can be seen from table 1, composition a, composition B, composition C, and composition D all have bacteriostatic effects on mycobacteria, wherein the bacteriostatic level of composition a is extremely sensitive, the level of composition B is highly sensitive, and the levels of composition C and composition D are medium sensitive or low sensitive, so that it can be seen that the composition containing juniper extract, dancing grass extract, and enrofloxacin has significant bacteriostatic effects on mycobacteria, wherein the bacteriostatic effect of composition a is particularly significant, while for composition B, when the composition is prepared, the bacteriostatic component in composition B cannot be sufficiently extracted by heating in water bath at a lower temperature of 45 ℃ so that the effect of composition B is greatly different from that of composition a, and for composition C, when the composition C is heated in water bath at a high temperature of 90 ℃ so as to evaporate bacteriostatic substances, which seriously affects the bacteriostatic effect of composition, and for composition D, in the process of preparing composition, the juniper extract and the dancing grass extract are directly mixed without the steps of water bath heating and dripping, temperature rise and fall are not changed, a large amount of antibacterial ingredients cannot be extracted, and the effect of the composition has a certain difference compared with the effect of the composition B. Therefore, the combination of the juniper extract, the dancing grass extract and the enrofloxacin in the composition has a remarkable bacteriostatic effect on mycobacterium, and the water bath heating in the mixing process also has a great influence on the bacteriostatic effect of the composition on mycobacterium.
Example 3 Effect of the mixture ratio of Juniperus Rigida extract and Dandelion extract on the bacteriostatic Effect
Accurately weighing 500g of juniper chinensis, drying, grinding into powder, adding the powder into 10 times of 60% ethanol, leaching for 2 hours, repeatedly leaching for 5 times, mixing leaching solutions, and concentrating to obtain juniper chinensis extract.
Weighing 5000g of dancing grass accurately, drying, grinding into powder, adding the powder into 10 times of 70% ethanol, leaching for 3 hours, and concentrating the leaching solution to obtain the dancing grass extract.
Heating 750g of the dancing grass extract in a water bath at 60 ℃ for 20 minutes, slowly dripping 40g of the juniper extract into the dancing grass extract, uniformly mixing to obtain a mixture, cooling the mixture to room temperature, then adding 1g of enrofloxacin into the mixture, and uniformly mixing to obtain the antibacterial composition A.
Heating 750g of the dancing grass extract in a water bath at 60 ℃ for 20 minutes, then slowly dripping 120g of the juniper extract into the dancing grass extract, uniformly mixing to obtain a mixture, cooling the mixture to room temperature, then adding 1g of enrofloxacin into the mixture, and uniformly mixing to obtain the antibacterial composition B.
Heating 80g of the dancing grass extract in a water bath at 60 ℃ for 20 minutes, slowly dripping 40g of the juniper extract into the dancing grass extract, uniformly mixing to obtain a mixture, cooling the mixture to room temperature, adding 1g of enrofloxacin into the mixture, and uniformly mixing to obtain the antibacterial composition C.
Heating 80g of the dancing grass extract in a water bath at 60 ℃ for 20 minutes, slowly and dropwise adding 20g of the juniper extract into the dancing grass extract, uniformly mixing to obtain a mixture, cooling the mixture to room temperature, adding 1g of enrofloxacin into the mixture, and uniformly mixing to obtain the antibacterial composition D.
Heating 750g of dancing grass extract in a water bath at 60 ℃ for 20 minutes, cooling to room temperature, adding 1g of enrofloxacin, and mixing uniformly to obtain the antibacterial composition E.
Heating 40g of juniper extract in a water bath at 60 ℃ for 20 minutes, cooling to room temperature, adding 1g of enrofloxacin, and uniformly mixing to obtain the antibacterial composition F.
Accurately weighing 1G of enrofloxacin, dissolving the enrofloxacin in 10 times of 60% ethanol, and uniformly mixing to obtain a composition G.
A bacterial suspension of mycobacteria was added to each tube to achieve a bacterial concentration of 105CFU/mL, each composition was diluted 1000-fold, and dilutions of composition a, composition B, composition C, composition E, composition F and composition G were added to different tubes, each was repeated 3 times without composition as a blank and without bacteria as a negative control. Shaking in a shaking table at 32 deg.C (180r/min) for 24h, taking out, comparing with a control tube, and observing, wherein the diameter of the inhibition zone and the inhibition grade are shown in Table 2.
TABLE 2 zone of inhibition diameter and grade of inhibition
| Item | Diameter of bacteriostatic circle (mm) | Grade of bacteriostasis |
| Composition A | 22.08±0.26 | +++ |
| Composition B | 19.59±0.35 | ++ |
| Composition C | 14.36±0.15 | + |
| Composition D | 13.96±0.32 | + |
| Composition E | 12.93±0.28 | + |
| Composition F | 8.36±0.42 | + |
| Composition G | 7.37±0.38 | + |
| Blank control | — | — |
| Negative control | — | — |
In the above tables, "+ + + + +" indicates extreme sensitivity, "+" indicates medium sensitivity or low sensitivity, and "-" indicates no sensitivity (no bacteriostatic effect).
As can be seen from table 2, each of composition a, composition B, composition C, composition D, composition E, composition F and composition G has bacteriostatic effect on mycobacteria, wherein the bacteriostatic rating of composition a is hypersensitive, the rating of composition B is hypersensitive, and the ratings of composition D, composition E (without juniper extract), composition F (without dancing grass extract) and composition G (enrofloxacin) are moderately or hypoallergenic. Therefore, the composition containing enrofloxacin, the juniper extract and the dancing grass extract has a remarkable bacteriostatic effect on mycobacteria, wherein the bacteriostatic effect of the composition A is particularly remarkable, while the bacteriostatic effect of the composition B is different from that of the composition A in a certain way, and the composition B is different from the composition A in that the adding amount of the juniper extract is 120g, so that the excessively high adding amount of the juniper extract can reduce the bacteriostatic effect of the composition; the bacteriostatic effect of the composition C is remarkably different from that of the composition A, and the composition C is different from the composition A in that the addition amount of the dancing grass extract is 80, so that the bacteriostatic effect of the composition can be reduced by too little addition amount of the dancing grass extract, and the influence of the dancing grass extract on the bacteriostatic effect of the composition is far greater than that of the juniper extract; the composition D is different from the composition A in that the addition amount of the dancing grass extract is 80, the addition amount of the juniper extract is 120g, and the antibacterial effect of the juniper extract is different from that of the composition A and the composition B, which shows that the addition amount of the juniper extract and the dancing grass extract needs to be within a certain range to realize the optimal antibacterial effect; for the composition E and the composition F, the compositions respectively added with the Chinese juniper extract or the juniper extract have certain bacteriostatic effect on mycobacterium, but the bacteriostatic effect is obviously different from that of the compositions containing enrofloxacin, the Chinese juniper extract and the Chinese juniper extract, and for the composition G, the bacteriostatic effect of single enrofloxacin on mycobacteria is lower than that of the compositions respectively added with the Chinese juniper extract and the Chinese juniper extract.
Example 4 bacteriostatic Effect of the feed having bacteriostatic action on Mycobacterium, Nocardia seriolae and Aeromonas schubertii
Accurately weighing 500g of juniper chinensis, drying, grinding into powder, adding the powder into 10 times of 60% ethanol, leaching for 2 hours, repeatedly leaching for 5 times, mixing leaching solutions, and concentrating to obtain juniper chinensis extract.
Weighing 5000g of dancing grass accurately, drying, grinding into powder, adding the powder into 10 times of 70% ethanol, leaching for 3 hours, and concentrating the leaching solution to obtain the dancing grass extract.
Heating 750g of the dancing grass extract in a water bath at 60 ℃ for 20 minutes, slowly dripping 40g of the juniper extract into the dancing grass extract, uniformly mixing to obtain a mixture, cooling the mixture to room temperature, then adding 1g of enrofloxacin into the mixture, and uniformly mixing to obtain the antibacterial composition.
Accurately weighing 2g of the antibacterial composition, and uniformly mixing the antibacterial composition with 1000g of outsourcing feed to obtain the feed with the antibacterial function.
And (3) experimental fish: healthy hybrid snakeheads (100-.
The experiment sets 7 groups of hybridized snakeheads, mycobacteria, Nocardia seriolae and Aeromonas schubertii to be respectively injected into 2 groups of experiment groups, 100 hybridized snakeheads are respectively stocked in each experiment group without injecting a bacteria control group. One group of mycobacterium after challenge, nocardia seriolae and aeromonas schubertii is selected and fed with feed with bacteriostatic function, and the mortality of each group is observed for 1 week. Three replicates per group.
After the injection of bacteria, acute death of test fishes occurs on the 2 nd day, the dead fishes have ascites, septicemia symptoms such as liver, spleen and kidney enlargement, viscera do not have nodule, viscera of the dead fishes on the 4 th day begin to have similar nodule symptoms of naturally-occurring fishes, and viscera nodule symptoms also occur in the 6 th day non-dead fishes after the autopsy.
The treatment results are shown in table 3.
TABLE 3 therapeutic results
As can be seen from table 3, the feed comprising the composition of enrofloxacin, juniper extract and dancing grass extract has significant bacteriostatic effect on sarcoidosis with the pathogenic bacteria mycobacterium, nocardia seriolae and aeromonas schubertii.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (10)
1. An application of feed with bacteriostatic function in preparing medicines for inhibiting pathogenic bacteria of sarcoidosis.
2. Use according to claim 1, characterized in that: the feed comprises 0.01-1% of the bacteriostatic composition.
3. A bacteriostatic composition, which is characterized in that: the antibacterial composition comprises the following components in a mass ratio of 1: 1-100: 100-1000 parts of enrofloxacin, juniper extract and dancing grass extract.
4. Bacteriostatic compositions according to claim 3, characterized in that: the antibacterial composition comprises the following components in a mass ratio of 1: 1-100: 100-1000 parts of enrofloxacin, juniper extract and dancing grass extract.
5. Bacteriostatic compositions according to claim 4, characterized in that: the mass ratio of the enrofloxacin, the juniper extract and the dancing grass extract is 1: 1-50: 500-1000; the mass ratio of the enrofloxacin, the juniper extract and the dancing grass extract is 1: 35-50: 620-800.
6. Bacteriostatic compositions according to claim 5, characterized in that: the juniper extract is prepared by the method comprising the following steps: drying Juniperus Rigida, grinding into powder, adding the powder into 10-20 times of 50-70% ethanol, leaching for 1-3 hr, repeatedly leaching for 3-5 times, mixing leaching solutions, and concentrating to obtain Juniperus Rigida Lindl extract.
7. Bacteriostatic compositions according to claim 5, characterized in that: the dancing grass extract is prepared by the method comprising the following steps: drying herba Trifolii Pratentis, grinding into powder, adding the powder into 10-20 times of 60-70% ethanol, leaching for 1-3 hr, and concentrating the leaching solution to obtain the extract.
8. A method of preparing a bacteriostatic composition according to any one of claims 3-7, characterized in that: heating the juniper extract in a water bath at 40-80 ℃ for 10-30 minutes, then slowly dripping the dancing grass extract into the juniper extract, uniformly mixing to obtain a mixture, cooling the mixture to room temperature, then adding enrofloxacin into the mixture, and uniformly mixing to obtain the antibacterial composition.
9. A method for preparing a bacteriostatic composition according to claim 8, characterized in that: the temperature of the water bath heating step is 50-65 ℃, and the time is 15-25 minutes.
10. A feed with antibacterial function is characterized in that: the feed comprises 0.01-1% of bacteriostatic composition; the antibacterial composition comprises the following components in a mass ratio of 1: 1-100: 100-1000 parts of enrofloxacin, juniper extract and dancing grass extract.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101999999A (en) * | 2009-09-01 | 2011-04-06 | 珈侬生化科技(中国)有限公司 | Method for preparing active extract of Juniperus sibirica and whitening and anti-ageing active cosmetic |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101999999A (en) * | 2009-09-01 | 2011-04-06 | 珈侬生化科技(中国)有限公司 | Method for preparing active extract of Juniperus sibirica and whitening and anti-ageing active cosmetic |
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| 刘敬余: "《植物百科全书 探索天下 学生版》", 31 May 2018, 北京教育出版社 * |
| 莫金凤: "复方中草药对杂交鳢生长、肉品质及抗细菌感染能力的影响", 《中国优秀硕士论文全文数据库农业科技辑》 * |
| 莫金凤等: "乌鳢源舒伯特气单胞菌生物学特性及其药物敏感性分析", 《水产学报》 * |
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