Application of chebulinic acid or chebulinic acid in preparation of drugs for resisting aquatic pathogenic bacteria

Abstract

The invention discloses application of chebulinic acid or chebulinic acid in preparation of drugs for resisting aquatic pathogenic bacteria. According to the invention, active components for resisting pathogenic vibrio in the fructus chebulae extract are separated, purified and identified by adopting an activity tracking mode, and the active components are identified as chebulinic acid and chebulagic acid. The chebulinic acid and the chebulinic acid are simple to prepare and small in toxic and side effects, and have a strong bacteriostatic effect, especially for inhibiting vibrio parahaemolyticus. Experimental results show that the 96-hour median lethal concentrations of chebulagic acid and chebulagic acid to fish are respectively 561.653 mg / L and 525.011 mg / L, which are 22.47 times and 21.00 times of the minimum inhibitory concentration 25mg / L for inhibiting vibrio parahaemolyticus, and the chebulagic acid and the chebulagic acid are safe to the fish, have broad-spectrum antibacterial effect, have inhibition effect on pseudomonas aeruginosa, aeromonas veronii, aeromonas hydrophila, vibrio alginolyticus, vibrio vulnificus, vibrio parahaemolyticus and the like, and can be used for preparing a compound preparation for treating the vibrio parahaemolyticus. Good application prospects are realized in the field of novel environment-friendly fishery medicines.

Description

Technical Field

[0001] The invention relates to the technical field of biomedicine, and in particular to application of chebulic acid or chebulic acid in preparing drugs for resisting aquatic pathogens. Background Art

[0002] Aquatic pathogenic microorganisms are the main source of aquatic product safety problems and diseases, which not only bring huge economic losses to global aquaculture, but also pose a threat to human health, especially some pathogenic Vibrio. In recent years, my country's aquaculture industry has developed rapidly, and fishery output has increased significantly, ranking among the top in the world. However, with the continuous increase in the scale and intensiveness of aquaculture, diseases caused by aquatic pathogens have occurred frequently, which not only affects the quality and safety of aquatic products, but also poses a challenge to the sustainable development of the aquaculture industry.

[0003] At present, chemical drugs are mostly used to prevent and treat aquatic bacterial diseases, such as antibiotics. The large-scale and long-term use of traditional antibiotics has produced a large number of drug-resistant strains, which not only increases the difficulty of bacterial disease prevention and control, but also poses a threat to food safety and public health. Chinese herbal medicine has become a potential substitute for antibiotics due to its advantages such as being green, safe, with little toxicity and side effects, and not prone to drug resistance. In addition, in order to reduce the use of antibiotics, the Ministry of Agriculture and Rural Affairs has promoted the use of veterinary antibiotics to reduce the amount of drugs used, and encouraged the use of veterinary Chinese medicine, microecological preparations and other green veterinary drugs without residues to replace some veterinary antibiotics, so as to promote the green development of the breeding industry and the safety of aquatic products.

[0004] Both chebulic acid and chebulic acid are derived from chebulic fruit. The structure of the compound has been disclosed in the literature (Gunawan-Puteri, Maria et al. Novel alpha-glucosidase inhibitors from Macaranga tanarius leaves. Food Chemistry, 2010, 123 (2): 384-389.). The structural formula of chebulic acid is:

[0005]

[0006] The molecular formula is: C 41 H 30 O 27 , molecular weight is 954.66, white amorphous powder. Easily soluble in organic solvents such as chloroform, methanol, acetone, ethanol, etc.

[0007] The structural formula of chebulic acid is:

[0008]

[0009] The molecular formula is: C 41 H32 O 27 , molecular weight is 956.68, white amorphous powder. Easily soluble in organic solvents such as chloroform, methanol, acetone, ethanol, etc.

[0010] Terminalia chebula Retz. and its variant Terminalia chebula Retz.var.tomentella Kurt. are dried and mature fruits of the Combretaceae plant Terminalia chebula Retz. and its variant Terminalia chebula Retz.var.tomentella Kurt., which are distributed in Tibet, Yunnan, Guangdong, Guangxi and other places in my country. Terminalia chebula has multiple pharmacological activities such as anti-oxidation, anti-inflammatory, anti-tumor, anti-pathogenic microorganism, hypoglycemic, anti-obesity, and prevention of cardiovascular diseases, and has great medicinal development value. There is no report that the active ingredients in Terminalia chebula, chebulic acid and chebulic acid, have anti-aquatic pathogenic bacteria effects. Summary of the invention

[0011] In order to overcome the shortcomings and deficiencies of the prior art, the present invention aims to provide an application of chebulic acid or chebulic acid in the preparation of drugs against aquatic pathogens. The chebulic acid and chebulic acid in the chebulic acid extract have low toxicity and side effects and have a strong antibacterial effect, especially in inhibiting Vibrio parahaemolyticus.

[0012] The present invention separates, purifies and identifies the active ingredients in the terminalia chebula extract that are effective against pathogenic Vibrio, and identifies them as terminalia acid and terminalia acid. Further studies have found that the active ingredients in terminalia chebula can inhibit a variety of aquatic pathogens.

[0013] The purpose of the present invention is achieved through the following technical solutions:

[0014] Use of chebulic acid or its pharmaceutically acceptable salt in the preparation of anti-aquatic pathogenic bacteria drugs or feed additives;

[0015] Use of chebulic acid or chebulic acid or a pharmaceutically acceptable salt thereof in the preparation of a medicine or feed additive for resisting aquatic pathogen infection;

[0016] The aquatic pathogenic bacteria is at least one of Vibrio parahaemolyticus, Vibrio alginolyticus, Vibrio vulnificus, Pseudomonas aeruginosa, Aeromonas vermifuge and Aeromonas hydrophila, and further Vibrio parahaemolyticus.

[0017] The chebulic acid or chebulic acid can be prepared by separation and extraction from plants, synthesis or semi-synthesis by various methods known in the art or alternative methods, or can be directly purchased commercially.

[0018] Furthermore, the chebulic acid or the chebulic acid is extracted and separated from chebulic fruit.

[0019] The medicine or feed additive comprises a therapeutically effective amount of chebulic acid or chebulic acid or a pharmaceutically acceptable salt thereof.

[0020] The MIC values ​​of the chebulic acid or chebulic acid against Vibrio parahaemolyticus are both 25 mg / L, and chebulic acid is safe to fish within the antibacterial concentration range of 25 mg / L to 143 mg / L, and chebulic acid is safe to fish within the antibacterial concentration range of 25 mg / L to 169 mg / L.

[0021] Furthermore, the safe concentration of chebulic acid is 143.21 mg / L, and the safe concentration of chebulic acid is 169.27 mg / L.

[0022] The medicine is prepared into liquid or powder.

[0023] The medicine contains one or more pharmaceutically acceptable carriers or excipients.

[0024] The chebulic acid and chebulic acid of the present invention can inhibit Vibrio parahaemolyticus, Vibrio alginolyticus, Vibrio vulnificus, Pseudomonas aeruginosa, Aeromonas vernix and Aeromonas hydrophila. Therefore, the antibacterial activity of chebulic acid and chebulic acid has a good application prospect in the field of new environmentally friendly fishery drugs.

[0025] Compared with the prior art, the present invention has the following advantages and effects:

[0026] (1) The present invention aims to find a new, highly effective and pollution-free drug resource for resisting aquatic pathogens in order to solve the current problem of Vibrio pathogens in aquaculture.

[0027] (2) The present invention is the first to use chebulic acid and chebulic acid to prepare drugs against aquatic pathogens, especially for inhibiting Vibrio parahaemolyticus.

[0028] (3) The raw materials of the method of the present invention are abundant. Terminalia chebula is a traditional Chinese medicine in my country and is easy to obtain. Terminalia chebula extract is obtained by using Terminalia chebula as a raw material and anhydrous ethanol as a solvent for extraction. It is determined that the effective ingredients of Terminalia chebula for inhibiting aquatic pathogens are chebulic acid and terminalia acid. In addition, terminalia acid and terminalia acid are easy to prepare and have broad application prospects.

[0029] (4) The MIC values ​​of chebulic acid and chebulic acid of the present invention against Vibrio parahaemolyticus are both 25 mg / L, and the 96h half lethal concentrations for fish are 561.653 mg / L and 525.011 mg / L, respectively, which are 22.47 and 21.00 times the minimum inhibitory concentration of 25 mg / L for inhibiting Vibrio parahaemolyticus. They are safe for fish and have a broad-spectrum antibacterial effect, and have inhibitory effects on Pseudomonas aeruginosa, Aeromonas vermifuge, Aeromonas hydrophila, Vibrio alginolyticus, Vibrio vulnificus, Vibrio parahaemolyticus, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 It is cheleic acid 13 C-NMR spectrum.

[0031] Figure 2 It is cheleic acid 1 H-NMR spectrum.

[0032] Figure 3 It is chebulic acid 13 C-NMR spectrum.

[0033] Figure 4 It is chebulic acid 1 H-NMR spectrum.

[0034] Figure 5 This is the HPLC chromatogram of cheleic acid.

[0035] Figure 6 This is the HPLC chromatogram of chebulic acid.

[0036] Figure 7 It is the antibacterial test of cheleic acid against Vibrio parahaemolyticus (mg / L).

[0037] Figure 8 OD values ​​of Vibrio parahaemolyticus after treatment with cheleic acid at different concentrations; Note: compared with before culture, *p<0.05, **p<0.01.

[0038] Fig. 9 This is the antibacterial test of chebulic acid against Vibrio parahaemolyticus (mg / L).

[0039] Fig.10 OD values ​​of Vibrio parahaemolyticus after treatment with chebulic acid at different concentrations; Note: compared with before culture, *p<0.05, **p<0.01.

[0040] Fig.11 This is the growth curve of Vibrio parahaemolyticus treated with cheleic acid.

[0041] Fig.12 This is the growth curve of Vibrio parahaemolyticus treated with chebulic acid.

[0042] Fig.13 is the AKP activity of Vibrio parahaemolyticus after treatment with cheleic acid; Note: compared with CK, *p<0.05, **p<0.01.

[0043] Fig.14 is the AKP activity of Vibrio parahaemolyticus after treatment with chebulic acid; Note: compared with CK, *p<0.05, **p<0.01.

[0044] Fig.15 These are the morphological changes of Vibrio parahaemolyticus under the action of different concentrations of cheleic acid; a: CK; b: 1 / 2MIC; c: 1MIC. DETAILED DESCRIPTION

[0045] The present invention is further described in detail below in conjunction with embodiments and drawings, but the embodiments of the present invention are not limited thereto.

[0046] As a preferred embodiment, the chebulic acid or chebulic acid has activity against aquatic pathogens Vibrio parahaemolyticus, Vibrio alginolyticus, Vibrio vulnificus, Pseudomonas aeruginosa, Aeromonas vermifuge and Aeromonas hydrophila.

[0047] As a preferred solution, the chebulic acid or chebulic acid is within the effective antibacterial concentration range of 25-143 mg / L, is safe for fish, and has the effect of resisting aquatic pathogen infection.

[0048] As a preferred embodiment, the chebulic acid or chebulic acid can be prepared by separation and extraction from plants, synthesis or semi-synthesis by various methods known in the art or alternative methods, or can be directly purchased commercially.

[0049] As a preferred embodiment, the chebulic acid or terminalia acid is obtained by extraction and separation from terminalia chebula fruit.

[0050] The specific preparation method is as follows:

[0051] 2kg of dried Terminalia chebula fruit was extracted twice with 20L 90% ethanol under reflux, each time for 2 hours. The extracts were combined and concentrated to obtain 440g of total extract. After the total extract was suspended with water (m:v=1g:1mL), it was extracted with petroleum ether and ethyl acetate in turn to obtain 110g of petroleum ether fraction and 280g of ethyl acetate fraction. After antibacterial experiments, it was determined that the ethyl acetate fraction was active. The ethyl acetate fraction was chromatographed on a HP20 macroporous resin column, and 10%, 20%, 30%, 40%, 50%, 70%, 90%, and 100% ethanol concentrations were used as the elution system for gradient elution, with 12L of eluent per level. According to the antibacterial activity results of each part of the macroporous resin column chromatography, the 20% ethanol eluate was subjected to preparative high performance liquid chromatography, with methanol: water (15%-60%) as the mobile phase for gradient elution for 90 min, a flow rate of 5 ml / min, and an eluent of 2 L to obtain active compound 1, which was identified as chebulic acid ( 13 C-NMR spectrum is shown in Figure 1 , 1 H-NMR spectrum is shown in Figure 2 The 30% ethanol eluate was subjected to preparative high performance liquid chromatography with methanol: water (20%-50%) as the mobile phase for gradient elution for 90 min at a flow rate of 5 ml / min and an eluent of 2 L to obtain active compound 2, which was identified as terminalia acid ( 13 C-NMR spectrum is shown in Figure 3 , 1 H-NMR spectrum is shown in Figure 4 ).

[0052] The prepared chebulic acid and chebulic acid can be detected by the following method:

[0053] Determination of the purity of chebulic acid and chebulic acid by HPLC

[0054] Weigh 1 mg of chebulic acid and chebulic acid accurately, place in a 1 mL injection bottle, add 1 mL of methanol, dissolve by ultrasonic oscillation, and determine the sample purity by normalization method. The liquid phase instrument is Agilent 1100 series; Welch Ultimate XB-C18, 4 mm × 250 mm, 5 μm is the chromatographic column; mobile phase A is an aqueous solution of 0.1% phosphoric acid solution by volume; mobile phase B is analytical grade methanol; gradient elution (0-30 min, 15% → 60% B); volume flow rate is 1 mL / min; column temperature is 30 ° C; injection volume is 10 μL; detection wavelength is 270 nm.

[0055] The HPLC chromatogram of cheleic acid is as follows: Figure 5 As shown in the results, there is only one single peak without other impurity peaks, that is, the purity of cheleic acid is relatively high and there are few impurities. The purity of cheleic acid reaches more than 98%.

[0056] The HPLC chromatogram of chebulic acid is as follows: Figure 6 As shown in the results, there is only one peak without other impurity peaks, that is, the purity of chebulic acid is relatively high and there are few impurities. The purity of chebulic acid reaches more than 98%.

[0057] Example 1: Determination of MIC values ​​of chebulic acid and chebulic acid against Vibrio parahaemolyticus

[0058] (1) Preparation of chebulic acid and chebulic acid solutions: 10 mg of chebulic acid and chebulic acid were weighed on a 1 / 10,000 balance, and dissolved in 1 mL of ultrapure water containing 0.5% DMSO to obtain a 10000 mg / L primary stock solution. 0.1 mL of the primary stock solution and 0.9 mL of ultrapure water were taken to obtain 1 mL of a 1000 mg / L secondary stock solution. Then, using a two-fold gradient dilution method, 1 mL of 500 and 250 mg / L solutions were prepared, and then using a 10-fold gradient dilution method, 1 mL of 100, 50, and 25 mg / L solutions were prepared. At this time, the DMSO content of the 1000 mg / L solution was 0.05%, and accordingly, the DMSO content of the 0 mg / L (control group) solution was also prepared to be 0.05%.

[0059] (2) Preparation of bacterial solution: Before the experiment, 100 μL of Vibrio parahaemolyticus strains in a glycerol tube in a -80°C refrigerator were inoculated into 10 mL of LB liquid culture medium and cultured at 28°C with shaking at 200 rpm for 12 h overnight activation to allow the strain to be in the late exponential growth phase. The absorbance of the bacterial solution at 600 nm was measured and the concentration of each bacterial solution was adjusted to 1×107 to 1×10 8 CFU / mL.

[0060] (3) In a sterile environment, 100 μL of diluted bacterial solution and 100 μL of prepared drug solution were added to a 96-well plate, respectively, to make the final concentrations 500, 250, 125, 50, 25, and 12.5 mg / L, respectively. Three parallel groups were set up for each experiment. The plates were placed in a 28°C incubator for 12 h. The bacterial solution without chebulic acid or chebulic acid was used as the negative control (CK), and the bacterial solution with antibiotics (ampicillin, final concentration of 2500 mg / L) was used as the positive control (A+). The absorbance value at 600 nm was measured, where MIC was the lowest concentration of chebulic acid and chebulic acid when the bacteria in the plate were invisible to the naked eye.

[0061] The results are as follows: Figure 7 and Figure 8 It can be seen that the MIC value of cheleic acid on Vibrio parahaemolyticus for 12 hours is 25 mg / L, which shows that cheleic acid has an inhibitory effect on the growth of Vibrio parahaemolyticus. Fig. 9 and Fig.10 It can be seen that the MIC value of chebulic acid on Vibrio parahaemolyticus for 12 hours was 25 mg / L, which showed that chebulic acid had an inhibitory effect on the growth of Vibrio parahaemolyticus.

[0062] Example 2: Growth curve of Vibrio parahaemolyticus by chebulic acid and chebulic acid

[0063] (1) Preparation of chebulic acid and chebulic acid solutions: According to the method of Example 1, 30 mL of 25, 50, 100 and 0 (control) mg / L solutions were prepared.

[0064] (2) Preparation of bacterial solution: According to the method of Example 1, the concentration of bacterial solution was 1×107 to 1×10 8 80mL of Vibrio parahaemolyticus bacterial solution with CFU / mL.

[0065] (3) In a sterile environment, 10 mL of diluted bacterial solution and 10 mL of prepared drug solution were added to test tubes to make the final concentrations 12.5 (1 / 2MIC), 25 (1MIC), and 50 (2MIC) mg / L, respectively. Three parallel groups were set up for each experiment. The culture was carried out at 28°C and 200 rpm for 24 h. Samples were taken every 2 h. The bacterial solution without chebulic acid or chebulic acid was used as a negative control. The absorbance at 600 nm was measured and the growth curve was drawn.

[0066] The results are as follows: Fig.11 and Fig.12 It can be seen that with the increase of the concentration of chebulic acid and chebulic acid, the growth of Vibrio parahaemolyticus gradually weakened. In the 1 / 2MIC chebulic acid or chebulic acid treatment group, the growth trend of Vibrio parahaemolyticus showed a significantly slower trend compared with the control group. In the 1MIC and 2MIC chebulic acid or chebulic acid treatment groups, the growth curve was flat and no obvious bacterial growth occurred.

[0067] Example 3: Antibacterial effects of chebulic acid and chebulic acid on other pathogens

[0068] (1) Preparation of chebulic acid and chebulic acid solutions: According to the method of Example 1, 1 mL of each solution of 5000, 2500 and 1250 mg / L was prepared.

[0069] (2) Preparation of bacterial solution: According to the method of Example 1, the concentration of Pseudomonas aeruginosa, Aeromonas vermifuga, Aeromonas hydrophila, Vibrio alginolyticus, Vibrio vulnificus or Vibrio parahaemolyticus was prepared to be 1×107-1×10 8 CFU / mL of bacterial solution 1mL.

[0070] (3) The inhibitory effect of chebulic acid and chebulic acid on pathogenic bacteria was determined by agar diffusion method. In a sterile environment, 0.1 mL of bacterial solution was taken with a pipette and evenly spread on a solid culture medium. When no liquid flowed, three evenly distributed holes were punched on the solid culture medium with a hole puncher (diameter 6 mm). Two of the holes were added with 100 μL of drug solution, and the other hole was added with 100 μL of liquid culture medium (without drug) as a control. Three parallel groups were set for each experiment. The culture dish was then placed in a constant temperature incubator at 28°C for 12 hours, and the diameter of the inhibition zone (mm) was measured and recorded. The results are shown in Tables 1 and 2.

[0071] Table 1 Diameter of inhibition zone of cheleic acid against other pathogens (mm)

[0072]

[0073] Table 2 Diameter of inhibition zone of chebulic acid against other pathogens (mm)

[0074]

[0075] From the results in Table 1 and Table 2, it can be seen that chebulic acid or chebulic acid has an inhibitory effect on Pseudomonas aeruginosa, Aeromonas vermifuga, Aeromonas hydrophila, Vibrio alginolyticus, Vibrio vulnificus, and Vibrio parahaemolyticus.

[0076] Example 4: Effects of chebulic acid and chebulic acid on the activity of alkaline phosphatase (AKP) of Vibrio parahaemolyticus

[0077] (1) Preparation of chebulic acid and chebulic acid solutions: According to the method of Example 1, 3 mL of each solution with concentrations of 25, 50, 100 and 0 (control) mg / L was prepared.

[0078] (2) Preparation of bacterial solution: According to the method of Example 1, the concentration of bacterial solution was 1×107 to 1×10 8 CFU / mL of Vibrio parahaemolyticus bacterial solution 50mL.

[0079] (3) In a sterile environment, add 1 mL of diluted bacterial solution and 1 mL of prepared drug solution to test tubes, respectively, to make the final concentrations 12.5 (1 / 2MIC), 25 (1MIC), and 50 (2MIC) mg / L, respectively. Set up three parallel groups for each experiment. Cultivate at 28°C and 200 rpm for 10 h. Use bacterial solution without chebulic acid or chebulic acid as negative control (CK). Take 1 mL of bacterial solution, centrifuge at 12000 rpm for 1 minute, take the supernatant, and use alkaline phosphatase (AKP) kit to determine AKP content. Repeat each sample three times.

[0080] The results are as follows: Fig.13 and Fig.14 It can be seen that after treatment with chebulic acid or chebulic acid, the extracellular AKP activity of Vibrio parahaemolyticus at different concentrations was significantly increased compared with the control group (p<0.01), and increased with the increase of concentration, indicating that chebulic acid or chebulic acid can destroy the integrity of the cell wall of Vibrio parahaemolyticus.

[0081] Example 5: Effect of cheleic acid on the morphology of Vibrio parahaemolyticus

[0082] (1) Preparation of cheleic acid solution: According to the method of Example 1, 1 mL of each of 12.5 (1 / 2 MIC), 25 (1 MIC) mg / L and 0 (control) mg / L solutions was prepared.

[0083] (2) Preparation of bacterial solution: According to the method of Example 1, the concentration of bacterial solution was 1×107 to 1×10 8 CFU / mL of Vibrio parahaemolyticus bacterial solution 2mL.

[0084] (3) Drug treatment: Place a layer of cellophane on the LB solid plate, spread 100 μL of bacterial solution with a sterile spreader, incubate in a 37°C incubator for 3 h, and then take out for use. Cut the cellophane into 1×1 cm pieces and soak them in solutions with 1 / 2 MIC, 1 MIC, and a control group of cheleic acid for 6 h. Set the cellophane sample without drug treatment as the control (CK).

[0085] (4) Fixation: Fix in 2.5% (V / V) glutaraldehyde solution for 1.5 h.

[0086] (5) Dehydration treatment: Dehydrate the samples in 30%, 50%, 70%, 90%, and 100% alcohol concentrations, respectively, for 10 minutes at each concentration.

[0087] (6) Drying: Dry in a low-temperature freeze dryer for 1 hour.

[0088] (7) After drying, take out the cellophane sample and stick it on the copper stage with double-sided tape. After gold spraying, observe and photograph it under a scanning electron microscope.

[0089] The results are as follows: Fig.15 As can be seen in a, the Vibrio parahaemolyticus cells not treated with cheleic acid present a typical Vibrio structure with a complete and smooth short rod appearance; Fig.15 As can be seen in b, when exposed to 1 / 2MIC cheleic acid, obvious depressions appeared on the cell surface and the morphology was abnormal. Fig.15 As can be seen from c in the figure, when the concentration is 1MIC, Vibrio parahaemolyticus cells show holes and lysis, with no complete cell structure and intracellular substances leaking out.

[0090] Example 6: Safety evaluation of chebulic acid and chebulic acid on goby

[0091] Preliminary experiments determined that the drug concentrations of chebulic acid and chebulic acid that caused all deaths of gobies within 24 hours were 1000 mg / L, and the concentrations that did not cause deaths within 96 hours were 100 mg / L.

[0092] On the basis of the preliminary experiment, five concentration groups were set up, namely 800, 500, 250, 100 and 0 (control) mg / L, with three parallels for each concentration. 27 buckets (volume: 10L) were filled with 2L of aerated 25‰ seawater and 8 gobies. The temperature was controlled at 23±0.5℃, and oxygen was supplied by an oxygen pump. During the experiment, the poisoning of gobies in each group was observed every day. If gobies were found to have died of poisoning, they should be fished out in time to avoid damaging the water quality and affecting the experiment. After 96h, the experimental results were statistically analyzed to calculate the median lethal concentration (LC50). 50) and 95% confidence interval (calculated using SPSS probit program), as shown in Table 3 and Table 4. Safety concentration = 48h LC 50 ×0.3 / (24h LC 50 / 48h LC 50 ) 2 .

[0093] Table 3 Results of acute toxicity test of chebulic acid and chebulic acid on goby

[0094]

[0095] Table 4 The median lethal concentration (LC50) of chebulic acid and chebulic acid on goby 50 )

[0096]

[0097] The results are as follows: From Table 3, it can be seen that the survival rate of gobies in the 100 and 0 mg / L treatment groups was 100%, and the mortality rate increased with the increasing drug concentration. In the 1000 mg / L treatment group, all gobies died within 24 hours. From Table 4, it can be seen that the 96h-LC of cheleic acid on gobies 50 The 96h-LC of chebulic acid in goby is 561.653mg / L, the safe concentration is 143.21mg / L. 50 The effective antibacterial concentration of the two active compounds is 525.011 mg / L, and the safe concentration is 169.27 mg / L. This shows that the two active compounds are safe for fish within the effective antibacterial concentration range of 25-143 mg / L and can be used in farmed fish.

[0098] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. Use of chebulic acid or its pharmaceutically acceptable salt in the preparation of anti-aquatic pathogen drugs or feed additives.

2. Use of chebulic acid or chebulic acid or a pharmaceutically acceptable salt thereof in the preparation of a medicine or feed additive for resisting aquatic pathogen infection.

3. The use according to claim 1 or 2, characterized in that: The aquatic pathogenic bacteria is at least one of Vibrio parahaemolyticus, Vibrio alginolyticus, Vibrio vulnificus, Pseudomonas aeruginosa, Aeromonas vermifuge and Aeromonas hydrophila.

4. The use according to claim 3, characterized in that: The aquatic pathogen is Vibrio parahaemolyticus.

5. The use according to claim 1 or 2, characterized in that: The chebulic acid or the chebulic acid is prepared by separation, extraction, synthesis or semi-synthesis from plants.

6. The use according to claim 5, characterized in that: The chebulic acid or the chebulic acid is obtained by extraction and separation from chebulic fruit.

7. The use according to claim 1 or 2, characterized in that: The safe concentration of chebulic acid is 143.21 mg / L, and the safe concentration of chebulic acid is 169.27 mg / L.

8. The use according to claim 1 or 2, characterized in that: The medicine is prepared into liquid or powder.

9. The use according to claim 1 or 2, characterized in that: The medicine contains one or more pharmaceutically acceptable carriers or excipients.

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