CN115869344A - Application of florfenicol and moringa oleifera leaf composition in preparation of medicine for preventing and treating acute hepatopancreatic necrosis of prawns - Google Patents
Application of florfenicol and moringa oleifera leaf composition in preparation of medicine for preventing and treating acute hepatopancreatic necrosis of prawns Download PDFInfo
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- 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
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Abstract
The invention discloses application of a florfenicol and moringa oleifera leaf composition in preparation of a medicine for preventing and treating acute hepatopancreatic necrosis of prawns, and belongs to the technical field of aquaculture. The florfenicol and the moringa leaves are prepared into a composition according to the mass ratio of 1: the composition can reduce vibrio density in Litopenaeus vannamei hepatopancreas infected with acute hepatopancreas necrosis; compared with the florfenicol and the moringa oleifera leaves which are used independently, the treatment effect of the composition on the artificial infection model of the acute hepatopancreas necrosis of the litopenaeus vannamei is obviously improved, the death rate of the litopenaeus vannamei after infection can be obviously reduced, the immunity of the litopenaeus vannamei is improved, and the hepatopancreas tissue structure of the infected litopenaeus vannamei is improved. Therefore, the invention can provide a high-efficiency, safe and low-toxicity pharmaceutical composition for preventing and treating the acute hepatopancreas necrosis of the prawns and provides a new direction for preventing and treating the acute hepatopancreas necrosis of the prawns.
Description
Technical Field
The invention relates to the technical field of aquaculture, in particular to application of a florfenicol and moringa oleifera leaf composition in preparation of a medicine for preventing and treating acute hepatopancreatic necrosis of prawns.
Background
Prawn acute hepatopancreatic necrosis syndrome (AHPND) is a newly discovered prawn epidemic disease in recent years, and can cause prawn hepatopancreatic atrophy, and the disease is considered to be mainly caused by vibrio parahaemolyticus at present. AHPND has great threat to the cultured shrimps, once infected with the shrimps, the death rate can reach 80 percent or even 100 percent, a large amount of cultured shrimps die in a short period, and the development of the prawn culture industry in China is seriously harmed. Therefore, the strengthening of the prevention and control of the AHPND of the prawns has important significance for the healthy cultivation of the seawater prawns and the safety of animal food. Chemical antibacterial drugs are the main means for controlling bacterial diseases in aquaculture industry, but long-term use of a large amount of chemical antibacterial drugs easily causes drug residues and drug-resistant bacteria in aquaculture environment and aquatic animals, and brings hidden troubles to prevention and control of aquaculture diseases and safety of aquatic animal-derived food. In addition, the public awareness of the danger of excessive exposure to chemical antibacterial agents is increasing, and people are also encouraged to search for more environmentally friendly and safer drugs. Under such circumstances, chinese herbal medicines are widely spotlighted because they are safer for aquatic animals and the environment than chemical antibacterial medicines. The chemical antibacterial agent and Chinese herbal medicine have different action mechanisms, and their combination can be used for treating many different diseases, and can reduce the dosage of chemical antibacterial agent. In view of the serious harm of AHPND to prawn culture, a chemical drug and a traditional Chinese medicine composition which can be effectively used for preventing and treating AHPND are urgently needed so as to reduce the use of chemical antibacterial drugs and the occurrence of drug resistance, which has important significance for prawn green culture.
Disclosure of Invention
The invention aims to provide application of a florfenicol and moringa oleifera leaf composition in preparation of a medicine for preventing and treating prawn AHPND (advanced high Performance Numbers disease) so as to solve the problems in the prior art.
In order to achieve the purpose, the invention provides the following scheme:
the invention provides application of a florfenicol and moringa oleifera leaf composition in preparation of a medicament for preventing and treating prawn AHPND.
The invention also provides application of the florfenicol and moringa oleifera leaf composition in preparing a medicine for preventing and treating prawn AHPND caused by vibrio parahaemolyticus.
The invention also provides application of the florfenicol and moringa oleifera leaf composition in inhibiting vibrio parahaemolyticus.
The invention also provides application of the florfenicol and moringa oleifera leaf composition in improving the sensitivity of the vibrio parahaemolyticus to the florfenicol.
Preferably, the mass ratio of florfenicol to moringa oleifera leaves in the composition is 1.
The invention also provides a medicament for preventing and treating the AHPND of the prawns, which comprises a florfenicol and moringa oleifera leaf composition, wherein the mass ratio of the florfenicol to the moringa oleifera leaves in the composition is 1.
The invention discloses the following technical effects:
in the composition disclosed by the invention, florfenicol is a third-generation chloramphenicol medicament, and plays a role in inhibiting the synthesis of bacterial protein by combining with a bacterial 50S ribosome subunit. The moringa leaves are used as a traditional Chinese herbal medicine with good drug effect, have no obvious toxic or side effect and residue, and are an ideal natural medicine. The florfenicol and moringa oleifera leaves are used in a combined mode, compared with the florfenicol or moringa oleifera leaves which are used independently, the death rate of prawns and the density of vibrio parahaemolyticus in hepatopancreatus are reduced remarkably, the activity of the immunoenzyme can be improved remarkably, the hepatopancreatus structure is improved, the hepatopancreatus tubule cavities are clearer, epithelial cells are arranged more neatly, and more complete cells exist. Therefore, the florfenicol and the moringa oleifera leaves are used in a combined manner, so that the drug effect can be enhanced, the dosage of the chemical antibacterial florfenicol is reduced, the problem of bacterial drug resistance caused by long-term use of a large amount of chemical antibacterial drugs in the culture production is avoided, the risk of safe production of food animals is reduced, and the food animals are more in line with green culture.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.
FIG. 1 is a graph of the effect of moringa leaves, florfenicol and drug combinations on AHPND infected Vibrio parahaemolyticus density in the hepatopancreat of Litopenaeus vannamei;
FIG. 2 is a graph of the effect of moringa leaves, florfenicol and drug combinations on AHPND infected litopenaeus vannamei mortality;
FIG. 3 is a graph of the effect of Moringa leaf, florfenicol and drug combination on AHPND infected litopenaeus vannamei immunoenzyme activity;
FIG. 4 is a graph of the effect of moringa leaves, florfenicol and drug combinations on the hepatopancreatic histopathology of AHPND infected litopenaeus vannamei.
Detailed Description
Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In addition, for numerical ranges in the present disclosure, it is understood that each intervening value, to the upper and lower limit of that range, is also specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in a stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference herein for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. The specification and examples are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.
Example 1 Effect of florfenicol and Moringa leaf composition on Vibrio parahaemolyticus Density in hepatopancreas of model of AHPND infection of Litopenaeus vannamei
1. Test materials
Vibrio parahaemolyticus strain 20130629002S01 (GenBank accession number CP 020036.1) capable of causing prawn to suffer from AHPND is separated and stored in a biological disease control and molecular pathology research laboratory of the yellow sea aquaculture institute. Florfenicol is available from Shanghai Aladdin Biotechnology Ltd (purity 98%, CAS No.: 73231-34-2), and Moringa oleifera leaves are available from Shandongya Biotechnology Ltd (purity greater than 99%, CAS No.: 93165-54-9).
2. Test method
The test is carried out in a 200L PVC plastic bucket, water is changed every day, the water temperature is kept at 28 +/-1 ℃, the pH value is 8.0 +/-0.1, and the salinity is 32 per mill. About 4.5g of healthy litopenaeus vannamei establishes an artificial infection model by injecting a vibrio parahaemolyticus strain 20130629002S01 into the abdomen. 150 shrimp were divided into 5 groups of 30 shrimp tails each, except for the blank controlAll the other 4 groups of Litopenaeus vannamei Boone are injected with 10 μ L of 10 7 CFU/mL of Vibrio parahaemolyticus. Immediately after infection, different groups of litopenaeus vannamei were fed with the feed containing florfenicol (16 mg/kg), moringa leaves (400 mg/kg) and the drug combination (8 mg/kg florfenicol +200mg/kg moringa leaves), and the blank control group and the infection-only group were fed with the blank feed once for 12 hours for 5 consecutive days. At days 0, 0.5, 1, 3 and 5 (d) post-infection, 6 shrimps were randomly taken from each group and dissected under sterile conditions to obtain the hepatopancreas. The obtained hepatopancreas were mixed and ground with 5mL of phosphate buffered saline (PBS, 0.1M, PH7.4). mu.L of the mixture was plated on thiosulfate-citrate-cholate-sucrose (TCBS) agar plates and after incubation at 28 ℃ for 24h, vibrio parahaemolyticus counting was performed.
As shown in FIG. 1, the density of Vibrio parahaemolyticus in the hepatopancreas of only the infection group, the moringa leaf group, the florfenicol group and the drug combination group decreased by 24.65%, 31.07%, 37.35% and 52.24% after 0.5 days, and by 47.75%, 52.27%, 58.59% and 71.65% after 1 day, respectively, throughout the entire experiment. 3 days after infection, the density of Vibrio parahaemolyticus in the hepatopancreas of only the infected group, the Moringa oleifera leaf group, the florfenicol group and the drug combination group was reduced to 30.63X 10 5 CFU/mL、26.59×10 5 CFU/mL、19.42×10 5 CFU/mL and 8.37X 10 5 CFU/mL. On day 5 post-infection, the pharmaceutical combination group prawn was almost completely cleared of vibrio parahaemolyticus in the hepatopancreas. The results show that: compared with the individual use of moringa leaves and florfenicol, the combined use of the moringa leaves and the florfenicol can obviously reduce the density of vibrio parahaemolyticus in the hepatopancreas of prawns infected by AHPND, obtain better anti-infection effect and obviously reduce the use amount of chemical antibacterial drugs of florfenicol.
Example 2 Effect of florfenicol and Moringa leaf compositions on mortality of an AHPND infection model of Litopenaeus vannamei
The strains, prawn feeding conditions, and treatment measures such as infection and administration according to this example are the same as those in example 1.
The 250-tailed shrimps were divided into 5 groups of 50 tails each. The test was repeated 3 times and the mortality was recorded at days 0, 0.5, 1, 3 and 5 (d) post infection.
The result is shown in figure 2, the cumulative mortality rate of litopenaeus vannamei after the litopenaeus vannamei is artificially infected with the vibrio parahaemolyticus is 82.65 percent within 5 days; mortality was 49.19% and 38.94% after 400mg/kg Moringa leaf and 16mg/kg florfenicol administration, respectively; mortality was 17.97% after administration of 8mg/kg florfenicol +200mg/kg moringa leaf composition. The research fully shows that compared with the single use of the moringa leaves and the florfenicol, the combined use of the moringa leaves and the florfenicol can obviously reduce the death rate of the AHPND infected litopenaeus vannamei, obtain better treatment effect and obviously reduce the use amount of the chemical antibacterial agent florfenicol.
Example 3 Effect of florfenicol and Moringa leaf compositions on the Immunity of an AHPND infection model of Litopenaeus vannamei
The strains, prawn feeding conditions, and treatment measures such as infection and administration in this example are the same as those in example 1.
The 450 shrimp tails were divided into 5 groups of 90 tails each. 18 shrimp and 6 shrimp were randomly sampled at days 0, 0.5, 1, 3 and 5 (d) post-infection, i.e., three samples per group were repeated at each sampling time. An equal volume of shrimp hemolymph was drawn using a syringe containing 0.2mL of anticoagulant. Plasma was separated by centrifugation at 3000g for 10 minutes and placed at-20 ℃ for subsequent determination of the immunoenzyme activity. The activities of superoxide dismutase (SOD), lysozyme (LZM), acid phosphatase (ACP) and alkaline phosphatase (AKP) in plasma were determined using a kit purchased from Nanjing institute for Biotechnology (Nanjing, china). And the protein content of the plasma sample is determined by a Bradford method by using a total protein quantitative kit of Nanjing institute of bioengineering and taking bovine serum albumin as a standard.
As shown in FIG. 3, after infection of Litopenaeus vannamei, the SOD, LZM, ACP and AKP activities of the group administered were all higher than those of the group infected only, and the immunoenzyme activities of the drug combination group (8 mg/kg florfenicol +200mg/kg Moringa oleifera leaves) were significantly higher than those of the florfenicol (16 mg/kg) and Moringa oleifera leaves (400 mg/kg) used alone. The results show that the combination of florfenicol and moringa leaves can obviously improve the immunity of the litopenaeus vannamei, and can obviously reduce the dosage of the chemical antibacterial florfenicol when a better immune effect is obtained.
Example 4 repair of florfenicol and Moringa oleifera leaf composition on hepatopancreas of Litopenaeus vannamei AHPND infection model
The strains, prawn feeding conditions, and treatment measures such as infection and administration according to this example are the same as those in example 1.
The 75-tailed shrimps were divided into 5 groups of 15 tails each. On day 5 (d), five groups of shrimps were dissected to obtain hepatopancreas, and after fixing the hepatopancreas samples for 24h using davison fixative, the hepatopancreas were sequentially placed in 70%, 80%, 90% ethanol solutions for 30 min each in ethanol of each concentration, and then placed in 95%, 100% ethanol for 2 times each for 20 min to dehydrate the hepatopancreas. The dehydrated hepatopancreas were embedded in paraffin, and cut using a conventional microtome to obtain hepatopancreas sections of 5 μm thickness, followed by Hematoxylin and Eosin (HE) staining. Histological changes of the hepatopancreas of the prawns in each test group were observed under an Olympus BX60 microscope.
As shown in FIG. 4, in the infection-only group, the epithelial cells of the hepatopancreas tubule of Litopenaeus vannamei were completely disrupted and disappeared, and in the florfenicol (16 mg/kg) and Moringa oleifera leaves (400 mg/kg) used alone, the cell morphology was improved and there were more intact cells. Compared with the single drug, in the drug combination group (8 mg/kg florfenicol +200mg/kg moringa oleifera leaves), the structural integrity and arrangement of the hepatopancreatic tubules are improved more, the lumens are clearer, and the number of the hepatopancreatic tubule epithelial cells is more and the arrangement is tighter. The result shows that the florfenicol and the moringa leaves are used in combination to remarkably improve the restoration capacity of the hepatopancreas tissues of the AHPND infected Litopenaeus vannamei, and the dosage of the florfenicol serving as a chemical antibacterial drug can be remarkably reduced when a better restoration effect is obtained.
The above-described embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solutions of the present invention can be made by those skilled in the art without departing from the spirit of the present invention, and the technical solutions of the present invention are within the scope of the present invention defined by the claims.
Claims (6)
1. The florfenicol and moringa oleifera leaf composition is applied to preparation of a medicine for preventing and treating acute hepatopancreatic necrosis of prawns.
2. The florfenicol and moringa oleifera leaf composition is applied to the preparation of the medicine for preventing and treating the acute hepatopancreatic necrosis of prawns caused by vibrio parahaemolyticus.
3. Use of florfenicol and moringa oleifera leaf composition for inhibiting Vibrio parahaemolyticus.
4. The application of the florfenicol and moringa oleifera leaf composition in improving the sensitivity of vibrio parahaemolyticus to florfenicol.
5. The use according to any one of claims 1 to 4, wherein the mass ratio of florfenicol to moringa oleifera leaves in the composition is 1.
6. The drug for preventing and treating the acute hepatopancreatic necrosis of prawns is characterized by comprising a florfenicol and moringa oleifera leaf composition, wherein the mass ratio of the florfenicol to the moringa oleifera leaves in the composition is 1.
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| CN106942112A (en) * | 2017-04-28 | 2017-07-14 | 山东农业大学 | A kind of disinfectant for preventing and treating the acute Hepatopancreatic necrosis disease of prawn that vibrio parahaemolytious triggers |
| KR101904371B1 (en) * | 2018-04-30 | 2018-10-08 | 대한민국 | Composition for treating or preventing Acute Hepatopancreatic Necrosis Disease of Shrimp |
| CN112716987A (en) * | 2020-12-31 | 2021-04-30 | 西藏自治区农牧科学院水产科学研究所 | Premixed preparation for preventing and treating intestinal inflammation disease of lasalopecuroide and preparation method thereof |
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| CN106942112A (en) * | 2017-04-28 | 2017-07-14 | 山东农业大学 | A kind of disinfectant for preventing and treating the acute Hepatopancreatic necrosis disease of prawn that vibrio parahaemolytious triggers |
| KR101904371B1 (en) * | 2018-04-30 | 2018-10-08 | 대한민국 | Composition for treating or preventing Acute Hepatopancreatic Necrosis Disease of Shrimp |
| CN112716987A (en) * | 2020-12-31 | 2021-04-30 | 西藏自治区农牧科学院水产科学研究所 | Premixed preparation for preventing and treating intestinal inflammation disease of lasalopecuroide and preparation method thereof |
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