Composition for treating large yellow croaker pyramidoniasis as well as preparation method and application thereof

Abstract

The invention relates to a composition for treating large yellow croaker pyramidoniasis as well as a preparation method and application thereof, and belongs to the technical field of aquaculture. The composition comprises the following components: curcumin, bile acid, beta glucan, bacillus subtilis freeze-dried powder, marine red yeast freeze-dried powder, vitamin C, vitamin E and ethylicin. When the feed additive is added into basal feed to feed large yellow croakers infected with the trypanosomiasis, the death rate of sick fishes can be effectively reduced, the polypide loading capacity is remarkably reduced, and pathological injuries of livers and spleens of the large yellow croakers are relieved. In addition, the composition can also improve the non-specific immunity of the fish body, improve the liver function, enhance the antioxidant capacity of the liver, and improve the lipase activity of the intestinal tract at the same time. Therefore, the composition can be used as a medicine or a feed additive for treating the large yellow croaker pyramidoniasis, has the advantages of small dosage and remarkable effect, and provides a new technical approach for solving the problem that the current large yellow croaker pyramidoniasis lacks an effective prevention and control means.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of aquaculture, and particularly relates to a composition for treating pyramidal worm disease of large yellow croaker as well as a preparation method and application thereof. BACKGROUND

[0003] Garlicin is the main sulfur active ingredient in garlic, which has broad-spectrum antibacterial, fungal and parasitic effects, and is widely used in food preservation, livestock and poultry feed additives, disease prevention and control and other fields. Studies have shown that garlicin can inhibit the cysteine protease of parasites, Ca 2+ Review:antimicrobial properties of allicin used alone or in combination with other medications. Folia Microbiol (Praha). 2020, 65: 451-465.). However, natural garlicin has poor stability, is difficult to mass-produce and store, and is easily oxidized by various oxidizing agents to lose activity. Ethyl garlicin is a biomimetic drug of garlicin, which is an ethyl homolog of garlicin, and is currently used as a broad-spectrum fungicidal biomimetic pesticide (Fu Xin, et al. Synergistic fungicidal effect of ethyl garlicin and amino oligosaccharin on bacterial fruit spot of melon. Tropical Agricultural Sciences, 2025, 45(1): 76-81). However, whether ethyl garlicin has a killing effect on aquatic animal parasites has not been reported in relevant literature and supported by data.

[0004] Curcumin is a plant polyphenol extracted from the rhizomes of the herb turmeric, commonly used as a food additive. Curcumin has multiple pharmacological effects, including antibacterial, anti-inflammatory, anticancer, and anti-diabetic effects. It has been proven to be effective in treating Trypanosoma cruzi infection. In vivo studies on mice infected with Trypanosoma cruzi showed that curcumin significantly reduced parasitemia and parasite load in tissues, improved survival rate, and improved pathological conditions (Sueth-Santiago V, et al. The effectiveness of natural Diarylheptanoids against Trypanosoma cruzi: cytotoxicity, ultrastructural alterations and molecular modeling studies. PLoS ONE 11(9). 2016, e0162926). Oral curcumin had moderate efficacy against Trypanosoma congolense and Trypanosoma evansi infections in mice (Molefe-Nyembe NI, et al. In vivo efficacy of Curcumin and Curcumin Nanoparticle in Trypanosoma congolense, Broden 1904 (Kinetoplastea: Trypanosomatidae)-infected mice. Pathogens. 2023, 12(10): 1227). In addition, curcumin has hepatoprotective, anti-inflammatory, antioxidant, and antitumor effects.

[0005] When large yellow croaker is infected with trypanosoma, the body surface is severely ulcerated, the liver is enlarged and whitish, the spleen is swollen and hemorrhagic, the intestinal contents are empty, and yellow mucous fluid often flows out. Therefore, when treating trypanosoma disease in large yellow croaker, it is necessary to treat the specimen and kill the insect at the same time, and improve the liver function and the ability of the digestive tract to digest food. SUMMARY

[0006] The purpose of the present application is to provide a composition for treating trypanosoma disease in large yellow croaker and its preparation method and application, in order to solve the problem that there is no effective prevention and control measure for trypanosoma disease in large yellow croaker at present.

[0007] To solve the above problems, the technical scheme adopted by the present application is as follows: The first aspect of the present application provides a composition for treating trypanosoma disease in large yellow croaker, which is composed of the following raw materials in parts by mass: curcumin 1-5 parts, bile acid 0.5-3 parts, beta glucan 1-8 parts, Bacillus subtilis freeze-dried powder 0.2-0.5 parts, Haematococcus pluvialis freeze-dried powder 0.1-0.25 parts, vitamin C 1-4 parts, vitamin E 0.5-2 parts, and ethiosom 2-8 parts. Further, the composition is composed of curcumin 1 part, bile acid 3 parts, beta glucan 6 parts, Bacillus subtilis freeze-dried powder 0.25 parts, Haematococcus pluvialis freeze-dried powder 0.25 parts, vitamin C 3 parts, vitamin E 0.5 parts, and ethiosome 6 parts by mass; Further, the number of viable bacteria in the Bacillus subtilis freeze-dried powder is 4×10 9 CFU / g, and the number of viable bacteria in the Haematococcus pluvialis freeze-dried powder is 5×10 9 CFU / g.

[0008] The second aspect of the present application provides the use of the above-mentioned composition in the preparation of a product for treating the spine worm disease of large yellow croaker; Further, the product includes a medicine and a feed additive; Further, the use method of the composition in the preparation of a medicine for treating the spine worm disease of large yellow croaker is as follows: curcumin is uniformly mixed with bile acid, beta glucan, Bacillus subtilis freeze-dried powder, Haematococcus pluvialis freeze-dried powder, vitamin C, and vitamin E to obtain mixture 1; ethiosome is uniformly mixed with corn starch, dried, and mixture 2 is obtained; mixture 1 and mixture 2 are uniformly mixed to obtain a medicine for treating the spine worm disease of large yellow croaker; and the medicine is administered orally. Further, the use method of the composition in the preparation of a feed additive for treating the spine worm disease of large yellow croaker is as follows: curcumin is uniformly mixed with bile acid, beta glucan, Bacillus subtilis freeze-dried powder, Haematococcus pluvialis freeze-dried powder, vitamin C, and vitamin E, and is added to feed raw materials to make granular feed; ethiosome is sprayed on the granular feed, mixed, and dried.

[0009] The present application has the following advantages: The in vivo experimental results of the present application show that the mortality of large yellow croaker infected with the spine worm disease can be reduced by 31.33% (the mortality of the control group is 50.23%) when 600 mg / kg of the composition medicine is added to the basic feed of large yellow croaker; and the mortality of large yellow croaker infected with the spine worm disease can be reduced by 23.66% and 25.92% when 200 mg / kg and 400 mg / kg of the composition medicine are added, respectively.

[0010] The present application researches and finds that adding the composition in the basic feed can reduce the pathological damage of the viscera of the Pseudosciaena crocea infected with the Trypanosoma and reduce the Trypanosoma load of the body. Specifically, the safe dose of the composition medicine added in the basic feed for the Pseudosciaena crocea is not more than 600 mg / kg, and the composition medicine with the dose of 600 mg / kg can significantly reduce the mortality of the Pseudosciaena crocea infected with the Trypanosoma. In addition, on the 6th day of feeding the composition medicine, the Trypanosoma load of the Pseudosciaena crocea is extremely significantly reduced, and the pathological damage of the liver and the spleen is also obviously improved. It is shown that the composition can be used for preparing a new medicine or a feed additive for treating the Trypanosoma infection of the Pseudosciaena crocea.

[0011] Meanwhile, the present application research also finds that adding the composition in the basic feed can repair the liver and intestinal damage and enhance the antioxidant capacity of the body. Specifically, when the composition medicine added in the basic feed is not more than 600 mg / kg, the activities of the serum glutathione (ALT) and the serum transaminase (AST) of the Pseudosciaena crocea are obviously reduced compared with those before the medicine is given and the basic feed control group, and the activity of the alkaline phosphatase (AKP) is obviously increased compared with those before the medicine is given and the control group. Further research shows that the activity of the superoxide dismutase (SOD) in the liver of the Pseudosciaena crocea is significantly increased, and the content of the malondialdehyde is significantly reduced. In addition, the activity of the intestinal lipase is also obviously improved.

[0012] In summary, adding the composition in the basic feed can reduce the mortality of the Pseudosciaena crocea infected with the Trypanosoma, reduce the Trypanosoma load, reduce the pathological damage of the liver and the spleen, and also improve the non-specific immunity of the fish body, improve the liver function, enhance the antioxidant capacity of the liver and improve the activity of the intestinal lipase. BRIEF DESCRIPTION OF DRAWINGS

[0013] Figure 1 Typical symptoms of the Pseudosciaena crocea infected with the Trypanosoma. A: large-area ulcer on the body surface; B: liver color changed to white.

[0014] Figure 2 Treatment effect of the medicine added in the basic feed on the Trypanosoma infection of the Pseudosciaena crocea. A: daily mortality rate; B: cumulative mortality rate; C: Trypanosoma load.

[0015] Figure 3 Influence of the medicine added in the basic feed on the morphological structure of the liver of the Pseudosciaena crocea. A: liver of the healthy fish; B: liver of the Pseudosciaena crocea before the medicine is given; C: liver of the Pseudosciaena crocea in the control group after the medicine is given; D: liver of the Pseudosciaena crocea in the 600 mg / kg medicine treatment group; E: liver of the Pseudosciaena crocea in the 400 mg / kg medicine treatment group; F: liver of the Pseudosciaena crocea in the 200 mg / kg medicine treatment group. The Pseudosciaena crocea in B-F are all infected with the Trypanosoma. The box represents the necrotic area of the liver, and the arrow represents the vacuolar degeneration of the hepatocytes (40x).

[0016] Figure 4: Effects of medicament added in basal diet on the morphological structure of the spleen of Pseudosciaena crocea. A: the spleen of healthy fish; B: the spleen of Pseudosciaena crocea before administration; C: the spleen of Pseudosciaena crocea in the control group after administration; D: the spleen of Pseudosciaena crocea in the 600 mg / kg medicament treatment group; E: the spleen of Pseudosciaena crocea in the 400 mg / kg medicament treatment group; F: the spleen of Pseudosciaena crocea in the 200 mg / kg medicament treatment group. The Pseudosciaena crocea in B-F were all infected with Trypanosoma sp. The arrow indicates the necrotic area of the spleen (40x).

[0017] Figure 5 : Effects of medicament added in basal diet at 600 mg / kg on the activities of several enzymes in the serum of Pseudosciaena crocea. A: alanine aminotransferase (ALT); B: aspartate aminotransferase (AST); C: alkaline phosphatase (AKP).

[0018] Figure 6 : Effects of medicament added in basal diet at 600 mg / kg on the activities of antioxidant enzymes in the liver of Pseudosciaena crocea. A: superoxide dismutase (SOD); B: catalase (CAT); C: malondialdehyde (MDA).

[0019] Figure 7 : Effects of medicament added in basal diet at 600 mg / kg on the activities of digestive enzymes in the intestine of Pseudosciaena crocea. A: amylase (AMS); B: trypsin; C: lipase.

[0020] Figure 8 : Effects of medicament on the recovery of liver function of Pseudosciaena crocea cultured in the fishing net in the Changyao sea area of Xiapu County, Ningde City, Fujian Province. A: the liver before administration was brittle, inelastic, and white in color; B: the liver after administration was obviously restored to purple red in color and elastic in texture. DETAILED DESCRIPTION

[0021] The application will be further described in conjunction with the examples. It should be noted that the following examples are only illustrative and are not intended to limit the protection scope of the application.

[0022] The technical solutions described in the application are all conventional technologies in the art if not specifically stated; the reagents or materials are all from commercial channels if not specifically stated. Among them, the number of viable bacteria in the freeze-dried powder of Bacillus subtilis is 4x10 9 CFU / g; the number of viable bacteria in the freeze-dried powder of marine Rhodovulum is 5x10 9 CFU / g; the CAS registration number of ethyl propyl disulfide is 682-91-7.

[0023] Example 1 A composition for treating big yellow croaker trypanosomiasis, consisting of the following raw materials in parts by mass: curcumin 1 part, bile acid 3 parts, beta glucan 6 parts, bacillus subtilis freeze-dried powder 0.25 parts, marine red yeast freeze-dried powder 0.25 parts, vitamin C 3 parts, vitamin E 0.5 parts, and ethiosom 6 parts.

[0024] A preparation method of a medicine for treating big yellow croaker trypanosomiasis, comprising the following steps: S1: uniformly mixing 1 part by mass of curcumin with 3 parts by mass of bile acid, 6 parts by mass of beta glucan, 0.25 parts by mass of bacillus subtilis freeze-dried powder, 0.25 parts by mass of marine red yeast freeze-dried powder, 3 parts by mass of vitamin C, and 0.5 parts by mass of vitamin E to obtain a mixture 1; S2: uniformly mixing 6 parts by mass of ethiosom with 85 parts by mass of corn starch, and drying at a temperature <60℃ to obtain a mixture 2; S3: uniformly mixing the mixture 1 of S1 with the mixture 2 of S2 to obtain the medicine for treating big yellow croaker trypanosomiasis.

[0025] Example 2: On September 20, 2024, in the fishing row of the coastal area of Jianjiang Town, Luoyuan County, Ningde City, Fujian Province, big yellow croakers infected with trypanosomes showed typical symptoms of large-area ulcers on the body surface and severe whitening of the liver (see Figure 1 ). In this experiment, big yellow croakers infected with trypanosomes with a body weight of 196.9±9.74g were selected as the research object, and were divided into 4 groups by random grouping method, with 3 parallel samples in each group, a total of 12 net cages (specification 4.0m×4.0m×4.0m), and 300 experimental fish were put into each net cage. Before administration, the fish were temporarily raised for 3 days to adapt to the environment. In the live experiment, different doses of the medicine obtained in Example 1 were added to the basic feed of each group in a mixed feeding manner. The specific addition amount was set as: 0 mg / kg (control group, i.e. no medicine added to the feed), 200 mg / kg, 400 mg / kg, and 600 mg / kg of feed. The fish were fed once a day at 17:00, with 1 kg of feed per net cage per day, and the administration was continued for 6 days. During the experiment, the number of dead big yellow croakers was recorded every day, and the daily mortality rate and the cumulative mortality rate were calculated (daily mortality rate = number of deaths on the day / total number of initial experiments, cumulative mortality rate = cumulative number of deaths / total number of initial experiments); at the same time, 3 surviving individuals were randomly selected from each group, and the average number of parasites in a single field of view was counted under a microscope.

[0026] After 6 days of continuous feeding, the fish were fasted for 24 h, and 9 fish from each group were randomly selected for sample collection: tail vein blood was drawn, and after standing at 4°C for 4 h, the supernatant was centrifuged at 4000 g for 10 min, and then transferred to a 2.0 mL cryogenic tube, frozen in liquid nitrogen, and stored at -80°C for serum liver physiological and biochemical index detection; liver and spleen tissues were removed and fixed with Boun's solution, and the next day, after being washed with running water, they were transferred to 70% alcohol for storage, for pathological section preparation; at the same time, liver and intestinal tissues (the intestinal contents were washed with pre-cooled physiological saline before being frozen) were frozen in liquid nitrogen and stored at -80°C for liver antioxidant index and intestinal digestive enzyme activity determination. The baseline sample collection before administration also followed the above procedure.

[0027] The pathological section preparation steps of the large yellow croaker liver and spleen are as follows: gradient alcohol dehydration: the fixed tissues were sequentially placed in 70% ethanol, 80% ethanol, 90% ethanol, 95% ethanol, and 100% ethanol (I, II) for 1 h each time; wax immersion treatment: first transparent in xylene I and xylene II for 30 min each, and then immersed in paraffin I and paraffin II for 1 h each time; embedding operation: the wax- immersed tissues were placed in an embedding box, liquid paraffin was injected, and the tissue direction (section upward) was adjusted, and the ice table was accelerated to cool until the wax block solidified; sectioning and slicing: the wax block was trimmed to expose the tissue section (1-2 mm edge was reserved), and a 5 μm slice was cut by a microtome; the slice was floated in a 45°C water bath to flatten (to avoid wrinkles), and then was scooped with a glass slide and placed in a 50°C oven for drying for 30 min to ensure firm adhesion; de-waxing and rehydration: the slice was sequentially immersed in xylene I and xylene II for 10 min each; and then was immersed in 100% ethanol (I, II), 95% ethanol, 90% ethanol, 80% ethanol, and 70% ethanol for 5 min each; staining process: immersed in hematoxylin staining solution for 4 min, and then washed with running water for 1 min for water blueing; then immersed in eosin staining solution for 4 min, and washed with running water for 1 min; dehydration and transparency: dehydrated in 70% ethanol, 80% ethanol, 90% ethanol, and 95% ethanol for 30 s each, and then in 100% ethanol (I, II) for 5 min each; then transferred to xylene I and xylene II for transparency for 10 min each; mounting and storage: add neutral balsam, cover with a glass slide (avoid air bubbles), and store the slice at room temperature in the dark.

[0028] Serum liver physiological and biochemical indicators were measured: Alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (AKP) were detected using kits from Shanghai Jianglai Biotechnology Co., Ltd., and the experimental procedures were strictly performed according to the product instructions. Liver antioxidant indicators and intestinal digestive enzyme activity were measured: Liver and intestinal samples were homogenized in pre-cooled 0.85% physiological saline buffer, centrifuged at 12000 rpm for 10 min at 4°C, and the supernatant was collected for later use. The activities of superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA) in the liver were measured using kits from Shanghai Jianglai Biotechnology Co., Ltd.; the activities of intestinal amylase (AMS), trypsin, and lipase were measured using kits from Nanjing Jiancheng Bioengineering Institute. All procedures were strictly performed according to the corresponding instructions.

[0029] The daily mortality rate of large yellow croaker is as follows: Figure 2 As shown in Figure A, the daily mortality rate of all four groups of experimental fish showed a decreasing trend. The daily mortality rates of the 200 mg / kg, 400 mg / kg, and 600 mg / kg drug treatment groups were significantly lower than those of the control group (P<0.05); there was no statistically significant difference in daily mortality rate among the three drug dosage groups (P>0.05), but the daily mortality rate of the 600 mg / kg treatment group was generally lower than that of the other two dosage groups.

[0030] The cumulative mortality rate of large yellow croaker is as follows Figure 2 As shown in Figure B, the control group fed only with the basal diet had the highest cumulative mortality rate, reaching 50.23% ± 5.31% by day 6. In contrast, the cumulative mortality rates of the 200 mg / kg, 400 mg / kg, and 600 mg / kg drug treatment groups were significantly lower (P < 0.05), at 26.57% ± 3.81%, 24.31% ± 3.24%, and 18.90% ± 1.79% respectively by day 6. No significant differences were observed among the three drug dosage groups (P > 0.05), but the cumulative mortality rate of the 600 mg / kg treatment group was significantly lower than that of the control group by 31.33%.

[0031] The amount of trypanosomes on the large yellow croaker is as follows Figure 2 As shown in C, the number of trypanosomes in all four groups of experimental fish showed a decreasing trend day by day. Compared with the control group, the number of trypanosomes in the 200 mg / kg, 400 mg / kg and 600 mg / kg drug treatment groups was significantly reduced (P<0.05), with the 600 mg / kg treatment group having the lowest number of trypanosomes, with only 1 out of 3 fish showing 1 trypanosome.

[0032] Effects of drugs on the morphology and structure of liver tissue in large yellow croaker, such as Figure 3 As shown. In a normal large yellow croaker, the liver cells are arranged neatly and orderly, with clearly distinguishable cell boundaries, and the nucleus is located in the center of the cell. Figure 3A); when the P. argentatus infected with T. breviabilis, the liver showed pale due to ischemia, texture became soft and lost elasticity, normal tissue structure completely disappeared, large area of liver tissue necrosis, most of the hepatocytes dissolved( Figure 3 B); after infection with T. breviabilis, the P. argentatus fed with basic feed only, the arrangement of liver cells was disorderly, the boundary between cells was unclear, most of the cells were vacuolar degeneration, part of the nucleus dissolved and disappeared with necrosis( Figure 3 C). In contrast, after infection with T. breviabilis, the P. argentatus fed with basic feed added with 200 mg / kg, 400 mg / kg and 600 mg / kg drug, the color of liver gradually recovered to purple red, the texture also gradually recovered elasticity. Histological observation showed that the liver tissue structure showed a normalizing trend, among which the liver tissue structure of 600 mg / kg drug treatment group basically recovered to normal, the cells arranged in order, the nucleus was centered and the nucleolus was clear( Figure 3 D); however, in 400 mg / kg drug treatment group( Figure 3 E) and 200 mg / kg drug treatment group( Figure 3 F), obvious vacuolar degeneration of liver cells could still be observed.

[0033] The effect of drug on the morphological structure of spleen of P. argentatus is shown in Figure 4 The spleen of normal P. argentatus is rich in lymphocytes and spleen cells, only mixed with a small amount of red blood cells( Figure 4 A); when the P. argentatus infected with T. breviabilis, the spleen appeared obvious swelling and accompanied by severe hemorrhage, a large number of red blood cells could be seen in the tissue section, and part of the area had spleen cell necrosis( Figure 4 B); after infection with T. breviabilis, the P. argentatus fed with basic feed only, there were still a large number of red blood cells in the spleen section, and the necrosis of part of the spleen cells continued to exist( Figure 4 C). In contrast, after infection with T. breviabilis, the P. argentatus fed with basic feed added with 200 mg / kg, 400 mg / kg and 600 mg / kg drug, the spleen tissue structure showed a recovery trend. Among them, the spleen of high dose group (600 mg / kg) was basically normal, but there was still bleeding, and the number of red blood cells in the section was more( Figure 4 D); the bleeding phenomenon of spleen of middle dose group (400 mg / kg) and low dose group (200 mg / kg) was reduced compared with the control group, but bleeding and necrosis area could still be observed( Figure 4 E, Figure 4 F).

[0034] The analysis results of serum physiological and biochemical indexes are shown in Figure 5ALT and AST are key indicators for evaluating the liver function status of organisms. The data showed that the activities of ALT and AST in serum of all test groups were significantly reduced after the test compared with those before the test (P < 0.05). Among them, the activities of ALT and AST of the large yellow croaker fed with basal diet after infection with Trypanosoma sp. (control group) also significantly decreased, which may be related to the fact that the samples detected after the test were all from surviving individuals; while the activities of ALT and AST of the large yellow croaker fed with basal diet supplemented with 600 mg / kg of the drug after infection with Trypanosoma sp. were more significantly reduced, indicating that the liver function status of the large yellow croaker in this group was better than that of the control group. Alkaline phosphatase (AKP) plays an important role in the immune system of organisms and can help clear foreign bodies invading the body. The detection results showed that there was no significant difference in the AKP activity of the control group before and after the test; while the AKP activity of the serum of the drug treatment group was significantly increased after the test, and was significantly higher than that of the control group at the same period. In summary, the above results show that the addition of the drug in the basal diet helps to protect the liver function of the large yellow croaker and can improve its serum non-specific immune level.

[0035] The detection results of liver antioxidant indicators are shown in Table 4. Figure 6 Superoxide dismutase (SOD) can effectively scavenge free radicals in the body, reducing the generation of oxidation products; catalase (CAT) can decompose hydrogen peroxide into water and oxygen, thereby avoiding its damage to cells. The detection results showed that there was no significant change in the activities of SOD and CAT of the control group before and after the test, while the activities of SOD and CAT of the liver of the large yellow croaker in the 600 mg / kg drug treatment group were significantly increased compared with those before the test (P < 0.05). Malondialdehyde (MDA) as the end product of lipid peroxidation, its content can directly reflect the degree of peroxidation of cytoplasmic membrane. The data showed that the MDA content of the liver of the control group and each drug treatment group was significantly reduced after the test compared with that before the test, and the reduction amplitude of the drug treatment group was more obvious; at the same time, the MDA content of the drug treatment group was significantly lower than that of the control group after the test (P < 0.05).

[0036] The analysis results of intestinal digestive enzyme activity are shown in Table 5. Figure 7 The addition of 600 mg / kg of the drug in the basal diet can significantly improve the lipase activity of the intestine of the large yellow croaker, but has no significant effect on the activities of amylase and trypsin. Since the digestion and absorption of nutrients depend on the mechanical movement of the digestive organs and the decomposition of digestive enzymes, the activity of fish digestive enzymes can directly reflect the body's ability to digest and absorb nutrients. It is speculated that the improvement of intestinal lipase activity may be related to the liposolubility of the drug — it may promote the large yellow croaker to secrete more lipase to assist itself in completing the digestion and absorption process, and at the same time, this is also conducive to improving the utilization rate of the large yellow croaker to other fatty acids in the feed.

[0037] Example 3: In mid-May 2025, some fish farms in Changyao sea area of Xiapu County, Ningde City, Fujian Province, raised 60-100 g of large yellow croaker, which suddenly broke out of trypanosomiasis. The water temperature at the time of onset was 23℃, and about 100 large yellow croaker died in a single net cage per day. The sick fish showed slow swimming and lack of energy. Examination of the sick fish showed no obvious abnormal symptoms on the body surface, but the liver was white (A) upon dissection, and microscopic examination showed 10-15 trypanosomes in a single field of view, thus trypanosomiasis was diagnosed. For the sick fish of the fish farm, the method of Example 2 was used to treat the fish with the drug: the drug was mixed into the basic feed at an addition amount of 600 mg / kg, and the fish were fed once a day. After 6 days of continuous feeding, the color of the liver of the large yellow croaker was obviously red (B), and no trypanosomes were found under a microscope. Figure 8 Figure 8

[0038] The above-described examples are part of the embodiments of the present application, but not all the embodiments. The detailed description of the embodiments of the present application is not intended to limit the claimed application, but only represents selected embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative labor are within the scope of protection of the present application.​​

Claims

1. A composition for treating Pseudokodakiaiamiyabei in Larimichthys crocea, characterized in that: The composition consists of curcumin 1-5 parts, bile acid 0.5-3 parts, beta glucan 1-8 parts, Bacillus subtilis freeze-dried powder 0.2-0.5 parts, Haematococcus pluvialis freeze-dried powder 0.1-0.25 parts, vitamin C 1-4 parts, vitamin E 0.5-2 parts, and ethiosome 2-8 parts by mass.

2. The composition of claim 1, wherein: The composition consists of curcumin 1 part, bile acid 3 parts, beta glucan 6 parts, Bacillus subtilis freeze-dried powder 0.25 parts, Haematococcus pluvialis freeze-dried powder 0.25 parts, vitamin C 3 parts, vitamin E 0.5 parts, and ethiosome 6 parts.

3. The composition of claim 1, wherein: The viable cell number in the Bacillus subtilis freeze-dried powder is 4 x 10 9 CFU / g, and the viable cell number in the marine Rhodovulum freeze-dried powder is 5 x 10 9 CFU / g.

4. Use of the composition according to any one of claims 1-3 in the preparation of a product for treating the spine worm disease of Pseudosciaena crocea.

5. Use according to claim 4, characterized in that: The product includes a medicine and a feed additive.

6. Use according to claim 5, characterized in that: The composition is used in the preparation of a medicine for treating the spine worm disease of Pseudosciaena crocea by mixing curcumin with bile acid, beta glucan, Bacillus subtilis freeze-dried powder, Haematococcus pluvialis freeze-dried powder, vitamin C and vitamin E to obtain mixture 1, mixing ethiosome with corn starch to obtain mixture 2, and mixing mixture 1 with mixture 2 to obtain the medicine for treating the spine worm disease of Pseudosciaena crocea.

7. Use according to claim 5, characterized in that: The medicine is administered orally.

8. Use according to claim 5, characterized in that: The composition is used in the preparation of a feed additive for treating the spine worm disease of Pseudosciaena crocea by mixing curcumin with bile acid, beta glucan, Bacillus subtilis freeze-dried powder, Haematococcus pluvialis freeze-dried powder, vitamin C and vitamin E, adding them to feed raw materials to make granular feed, spraying ethiosome on the granular feed, mixing and drying.

Images