Application of Usnea powder, Usnea acid, or sodium usneate in the preparation of broad-spectrum antiparasitic drugs or feeds for aquatic animals

Abstract

This invention provides the application of Usnea powder, Usnea acid, or sodium usneate (or Usnea powder, Usnea acid, or sodium usneate as the sole active ingredient) in the preparation of broad-spectrum antiparasitic drugs for aquatic animals, drugs for treating complications caused by parasitic infections, feed additives, or functional feeds. By utilizing Usnea, Usnea extract, Usnea acid, and Usnea salts, various parasites, including ciliates, worms, and protozoa, that infest the surface and interior of aquatic animals during aquaculture can be eliminated or killed, thus maintaining the health of farmed animals, improving the success rate of aquaculture, achieving broad-spectrum and highly effective prevention and control of aquatic animal parasitic diseases, reducing the use of chemical drugs, and providing good aquaculture, ecological, and social benefits. It is safe, broad-spectrum, and highly effective, facilitating large-scale promotion.

Description

Technical Field

[0001] This invention relates to the field of veterinary drug technology, specifically to the application of Usnea powder, Usnea acid, or sodium Usnea in the preparation of broad-spectrum antiparasitic drugs or feeds for aquatic animals. Background Technology

[0002] my country is a major aquaculture country, and its development has been particularly rapid over the past 20 years. The aquaculture industry has not only seen a continuous increase in the types of farmed animals, but also a gradual increase in farming area and stocking density. Furthermore, the exchange of aquatic animals between regions and internationally has become increasingly frequent. At the same time, disease problems that constrain the sustainable development of aquaculture are becoming increasingly prominent, manifested in an increase in the types of diseases, more frequent outbreaks, faster and wider disease transmission, greater harm, higher mortality rates after onset, and severe economic losses.

[0003] Parasitic diseases in aquatic animals are a common category of illnesses, occurring in almost all farmed species. Different types of parasites have different living and reproductive habits, resulting in varying degrees of damage to aquatic animals. Parasitic diseases occur in waves, almost throughout the entire farming process from larvae to adults. Common aquatic animal parasites include: flagellates such as *Cryptocetus gillii*; ciliates such as *Trichodina*, *Chilodonella*, *Scutiformis*, *Ichthyophthirius multifiliis*, and *Cryptocetus irritans*; worms such as *Dactylogyrus* and *Gyrodactylus*; protozoa such as *Myxospora*, *Cordyceps*, and *Eimeria*; and arthropods such as *Anchor worms*, *Sinozoa*, copepods, and fish lice. These parasites reside in the gills and body surface of farmed animals, and some species reside in the digestive tract, causing damage, inflammation, and bleeding. Severe cases can lead to mass mortality. Furthermore, the damage caused by parasites easily triggers secondary infections by bacteria and fungi, resulting in bacterial and fungal diseases and causing even more serious harm. In some provinces of my country, especially in the south, fish parasitic diseases occur almost every year, causing fish deaths. The mortality rate is as low as 20% to 30%, and in severe cases it can reach more than 90%.

[0004] Treatment of parasitic infections in aquatic animals is challenging. Treatment typically involves various antiparasitic drugs, such as mebendazole, praziquantel, albendazole, and trichlorfon for helminthiasis; copper sulfate and ferrous sulfate for flagellates and ciliates; chlorpheniramine hydrochloride and diclazuril for protozoan parasites; and trichlorfon for arthropod parasites. However, due to years of weak research and development in antiparasitic drugs, effective drugs are scarce, and parasitic pathogens have developed strong resistance to existing drugs. This increases treatment costs, and excessive use leads to environmental pollution and drug residues, severely impacting the quality of farmed animal products.

[0005] Traditional Chinese medicine and its active ingredients, as new types of fish medicines, are characterized by safety, effectiveness, low toxicity, and no residue, and have great potential for development.

[0006] Usnea, also known as lady's wisteria, cloud grass, and old man's beard, is the lichen of *Usnea longifolia* and *Usnea ringifolia*, belonging to the Usneaceae family and the *Usnea* genus. Its properties are bitter, sweet, and neutral. Its functions include expectoration, liver clearing, detoxification, and hemostasis. It is used to treat cough with excessive phlegm, pulmonary tuberculosis, malaria with phlegm, headache, red eyes, eye swelling and boils, mastitis, traumatic bleeding, metrorrhagia, leukorrhea, rheumatic pain, and burns. Its application has a long history, recorded in the traditional medicine books of Mongolian, Tibetan, and Uyghur ethnic minorities. The medicinal materials used by various ethnic groups and regions all utilize the branched lichen of the *Usnea* genus, with *Usnea longifolia* being the most common. There is considerable research on the use of usnic acid in Usnea. The application of sodium usnicate is similar to that of usnic acid, focusing on its antibacterial, anti-infective, detoxifying, antitumor, and cosmetic effects. However, there are no reports on the application of Usnea, Usnea extract, usnic acid, or usnicate salts in the prevention and treatment of parasitic diseases in aquatic animals. Summary of the Invention

[0007] Purpose of the invention

[0008] The purpose of this invention is to provide the use of Usnea powder, Usnea acid, or sodium Usnea (or Usnea powder, Usnea acid, or sodium Usnea as the sole active ingredient) in the preparation of broad-spectrum antiparasitic drugs for aquatic animals or drugs or feed additives or functional feeds for complications caused by parasitic infections.

[0009] This invention utilizes aquatic animals suffering from parasitic diseases for drug screening, obtaining broad-spectrum and effective Usnea, Usnea extract, and their active ingredients, usnic acid and usnicate. The drugs of this invention have good anthelmintic or insecticidal effects on common aquatic ciliates, worms, and protozoa. Compared with traditional aquatic insecticides such as copper sulfate and ferrous sulfate powder, mebendazole, chlorpheniramine hydrochloride, and trichlorfon, they have advantages such as high safety, low drug toxicity and side effects, and no harmful residues. They can improve the quality and safety of aquatic products and have good ecological, environmental, and social benefits.

[0010] Solution

[0011] To achieve the objectives of this invention, the technical solution adopted is as follows:

[0012] In a first aspect, the present invention provides the use of Usnea powder, Usnea acid, or sodium usneate in the preparation of a drug, feed additive, or functional feed for broad-spectrum antiparasitic treatment of aquatic animal parasites or complications caused by parasite infection.

[0013] Secondly, the present invention provides the use of Usnea powder, Usnea acid, or sodium usne as the sole active ingredient in the preparation of a drug or feed additive or functional feed for broad-spectrum antiparasitic treatment of aquatic animal parasites or complications caused by parasite infection.

[0014] Thirdly, the present invention provides a broad-spectrum antiparasitic drug for aquatic animals, comprising Usnea pulverized material, and / or Usnea extract, and / or Usnea acid, and / or Usnea salt, wherein the active ingredients include Usnea acid and Usnea salt, wherein the amount of Usnea acid and / or Usnea salt is not less than 0.2% by mass, optionally not less than 1%, and optionally not less than 2%.

[0015] Fourthly, the present invention provides a broad-spectrum feed additive for combating aquatic animal parasites, comprising pulverized Usnea, and / or Usnea extract, and / or Usnea acid, and / or Usnea salt, wherein the active ingredients include Usnea acid and Usnea salt, wherein the amount of Usnea acid and / or Usnea salt is not less than 0.002% by mass, optionally not less than 0.02%, and optionally not less than 0.2%.

[0016] Fifthly, a broad-spectrum functional feed for combating aquatic animal parasites is provided, comprising pulverized Usnea, and / or Usnea extract, and / or Usnea acid, and / or Usnea salt, wherein the active ingredients include Usnea acid and Usnea salt, wherein the content of Usnea acid and / or Usnea salt is not less than 0.0002% by mass, optionally not less than 0.002%, and optionally not less than 0.02%.

[0017] In the first to fifth aspects mentioned above, the parasites are parasites that infect fish or crustaceans.

[0018] In the first to fifth aspects mentioned above, the parasites include one or more of ciliates, helminths, and protozoa; optionally, the ciliates include one or more of Trichodina (e.g., Trichodina disease in perch), Ichthyophthirius multifiliis (e.g., Ichthyophthirius multifiliis in carp), Chilodonella (e.g., Chilodonella multifiliis in yellow catfish), Cryptocaryon irritans (e.g., Cryptocaryon irritans in large yellow croaker), Shield ciliates (e.g., Shield ciliates in turbot), crustacean ciliates, Vorticella, Vorticella, Cyclodactylus, and Monotyphimurium; optionally, the helminths include one or more of Dactylogyrus (e.g., Dactylogyrus rufiformis in crucian carp), tapeworms (e.g., Tapeworm in grass carp), and Gyrodactylus; optionally, the protozoa include one or more of Sporozoa (e.g., Sporozoa rufiformis in crucian carp) and vertebral worms.

[0019] In the first to fifth aspects mentioned above, the drug is an oral drug or a medicated bath preparation; optionally, the drug contains not less than 0.2% by mass of usnic acid and / or usnic acid salt, optionally not less than 1%, optionally not less than 2%.

[0020] In the first to fifth aspects mentioned above, the feed additive contains not less than 0.002% by mass of isocyanate and / or isocyanate.

[0021] In the first to fifth aspects mentioned above, the functional feed contains not less than 0.0002% by mass of usnic acid and / or usnic acid salt.

[0022] In the first to fifth aspects mentioned above, the Usnea powder is Usnea pulverized material or Usnea extract; optionally, the Usnea extract is an organic solvent extract, and optionally the organic solvent includes one or more of acetone, chloroform, and glacial acetic acid.

[0023] In the first to fifth aspects mentioned above, the dosage form of the drug is any one of powder, granule, solution, ointment, pourable, or injection.

[0024] In the first to fifth aspects mentioned above, the drug also contains pharmaceutically acceptable excipients.

[0025] In the first to fifth aspects mentioned above, aquatic animals include fish and / or crustaceans; optionally, aquatic animals include one or more of the following: grass carp, common carp, crucian carp, perch, yellow catfish, catfish, large yellow croaker, turbot, golden pomfret, tilapia, whiteleg shrimp, giant freshwater prawn, tiger prawn, Japanese freshwater prawn, Chinese shrimp, Chinese mitten crab, and mud crab.

[0026] Beneficial effects

[0027] In this invention, various parasites, including ciliates, worms, and protozoa, can be eliminated or killed by using drugs (veterinary drugs), feed additives, and functional feeds made from Usnea, Usnea extract, Usnea acid, and Usnea salts during the aquaculture process. This helps maintain the health of farmed animals, improve the success rate of aquaculture, and achieve broad-spectrum and highly effective prevention and control of aquatic animal parasitic diseases. It reduces the use of chemical drugs, has good aquaculture, ecological, and social benefits, and is safe, broad-spectrum, and highly effective, making it suitable for large-scale promotion. Attached Figure Description

[0028] One or more embodiments are illustrated by way of example with reference to the accompanying drawings, and these illustrative examples are not intended to limit the embodiments. The term "illustrative" as used herein means "serving as an example, embodiment, or illustration." Any embodiment illustrated herein as "illustrative" is not necessarily to be construed as superior to or better than other embodiments.

[0029] Figure 1Test Example 2 of this invention shows the effects of using Usnea extract powder on Trichodina disease (cilia) in sea bass before and after application; where A is the situation before using Usnea extract powder, where a large number of Trichodina parasites can be seen on the gills; B is the situation after using Usnea extract powder for 3 days, where the number of Trichodina parasites on the gills is significantly reduced.

[0030] Figure 2 Test Example 3 of this invention describes the effects of using Usnea Sulfate Powder on grass carp tapeworm disease (a type of worm) before and after its application. In this example, A represents the situation before the application of Usnea Sulfate Powder, where a large number of tapeworms parasitize the intestines of grass carp. In B, after using Usnea Sulfate Powder for 5 days, the number of tapeworms parasitizing the intestines of grass carp decreased to 0.

[0031] Figure 3 The test example 3 of this invention shows the condition of crucian carp before the use of sodium usnic acid powder. Among them, a large number of diseased crucian carp fry died every day, and their throats and subcutaneous tissues were swollen with a large number of cysts.

[0032] Figure 4 The results of microscopic examination of crucian carp lesions before sodium usnic acid powdering in Test Example 3 of this invention showed a large number of spores.

[0033] Figure 5 The following are the conditions and microscopic examination results of crucian carp on the 5th day after using sodium usnic acid powder in Test Example 3 of this invention. Among them, A is a photo of the pond downwind, with basically no dead fish floating; B and C are subcutaneous photos of crucian carp, with no sporozoites found; D is a photo of the throat and gills, with no sporangia found; E is a microscopic photo, with no sporozoites found. Detailed Implementation

[0034] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0035] Furthermore, to better illustrate the present invention, numerous specific details are provided in the following detailed embodiments. Those skilled in the art should understand that the present invention can be practiced without certain specific details. In some embodiments, materials, elements, methods, and means well known to those skilled in the art are not described in detail in order to highlight the spirit of the invention.

[0036] Unless otherwise expressly stated, throughout the specification and claims, the term "comprising" or its variations such as "including" or "comprises" shall be understood to include the stated elements or components without excluding other elements or other components.

[0037] The Usnea of ​​this invention is commercially available.

[0038] Example 1: Matsutake Powder

[0039] Dry the Usnea medicinal material, grind it into a fine powder of 80 mesh, add an appropriate amount of glucose, and prepare a powder with a Usnea acid content of 2%, which is called Usnea Powder, for later use.

[0040] Example 2: Usnea extract powder

[0041] After drying the Usnea, weigh 10g, use 300ml of acetone as solvent, heat and extract, collect the extract, recover the solvent, concentrate the extract, add an appropriate amount of starch, dry and pulverize into powder, so that the Usnea acid content is about 2%, which is the Usnea extract powder, for later use.

[0042] Example 3: Usnea Sinensis Powder

[0043] Add an appropriate amount of glucose to the isoflavone acid to prepare a powder with an isoflavone acid content of 2%, which is the isoflavone acid powder, for later use.

[0044] Example 4: Sodium Usnicotinate Powder

[0045] Sodium usnic acid is mixed with an appropriate amount of glucose to prepare a powder with a sodium usnic acid content of 2%, which is called sodium usnic acid powder, for later use.

[0046] Weigh out a certain amount of the above-mentioned drugs and mix them with the feed step by step to make medicated feed for later use.

[0047] Test Comparison Example 1:

[0048] The applicant previously discovered that Usnea had a good effect on chicken coccidiosis, so they attempted to use the drug described in Example 3 to prevent and treat coccidiosis (Eimeria) in aquatic animals, but it did not show ideal therapeutic effects. Specifically:

[0049] An oral treatment experiment was conducted on 1-year-old grass carp infected with Eimeria coccidia. An 8-mu grass carp pond in Yangqiao Town, Guanyun District, Lianyungang, Jiangsu Province, was stocked with 900 1-year-old grass carp fingerlings per mu. In June, grass carp mortality occurred. Ten diseased fish were collected and dissected. Numerous small white nodules were found on the intestinal wall. Tissue samples from these nodules were prepared into slides and observed under a microscope, revealing coccidia oocysts. The diagnosis of Eimeria coccidia was confirmed. Statistics showed an average of 12 small white nodules per intestinal wall per fish. Approximately 30 grass carp died daily in the pond.

[0050] Treatment method: The *Usnea sulforaphane* powder from Example 3 was mixed evenly with the grass carp feed, and a binder was added to prepare a medicated feed with a *Usnea sulforaphane* powder content of 10 g / kg of feed. After feeding this feed continuously for 7 days, 10 grass carp were randomly selected, and the number of small nodules on the intestinal wall was observed using the same method. Simultaneously, the mortality rate of the grass carp after medication was observed to evaluate the therapeutic effect of *Usnea sulforaphane* powder on *Eimeria tenella* disease in grass carp. Results: After 7 days of use, the average number of small nodules on the intestinal wall of the grass carp increased from 12 / intestine to 15 / intestine. At the same time, the mortality rate of grass carp increased from approximately 30 fish per day to approximately 80 fish per day, with no significant improvement in symptoms. This indicates that *Usnea sulforaphane* powder from Example 3 did not show an ideal therapeutic effect on *Eimeria tenella* disease in grass carp.

[0051] The inventors analyzed that although chicken coccidia and fish coccidia both belong to the coccidia class of parasites, their life habits and drug susceptibility can vary significantly by species, making the development of drugs for parasites still very challenging. Through extensive experimentation and research, the applicant ultimately discovered that Usnea has good preventative and therapeutic effects against a wide range of parasites, including some protozoa, some ciliates, and some worms, demonstrating unexpected therapeutic effects. Specifically:

[0052] Test Example 1: The effect of using Usnea Powder (Example 1) on crucian carp with Dactylogyrus disease (a type of worm).

[0053] A total of 20 mu (approximately 3.3 hectares) of crucian carp farming ponds in Caofeidian District, Tangshan, Hebei Province, with a stock of 800 fish per mu, was randomly harvested on September 20. The gill filaments of the fish were cut off, pressed into tablets, and observed under a microscope (10×10 magnification). It was found that a large number of Gyrodactylus parasites were found on the gills of the crucian carp. The total number of Gyrodactylus parasites on the gill filaments was 28-32 per fish on average.

[0054] Treatment method: Mix the pine needle powder from Example 1 with crucian carp feed evenly, add a binder, and prepare a medicated feed with a pine needle powder content of 10g / kg of feed. After feeding continuously for 5 days, randomly select 10 crucian carp and observe the number of Gyrodactylus parasites on the gills of the crucian carp using the same method. At the same time, observe the health status, behavioral characteristics and feeding of the crucian carp after medication to evaluate the therapeutic effect of pine needle powder on crucian carp Gyrodactylus disease.

[0055] Results: After 5 days of use, the average number of Gyrodactylus parasites on the gills of crucian carp decreased from 28-32 per fish to 2-3 per fish. Simultaneously, the feeding speed of the crucian carp significantly increased; the time to finish feeding the same amount of feed was shortened from 2 hours before use to 1.5 hours. This demonstrates that the *Usnea indica* powder in Example 1 has a good therapeutic effect on Gyrodactylus infection in crucian carp.

[0056] Test Example 2: The effect of Usnea extract powder (Example 2) on Trichodina disease (cilia) in sea bass.

[0057] A 25-mu (approximately 4.3 hectares) sea bass pond in Yueyang, Hunan Province, with a stock of 50,000 fish, began to feed less well in mid-June, exhibiting signs of swarming. Suspecting parasitic infestation, five sea bass were randomly selected, and gill filaments were collected, pressed into slides, and examined under a microscope (10×10x). Numerous Trichodina parasites were found on the gills of the sea bass. A count revealed an average of 40-50 Trichodina parasites per field of view (e.g., ...). Figure 1 A).

[0058] Treatment method: Dissolve and dilute the Usnea extract powder from Example 2 in an appropriate amount of water and then spray it throughout the pond. The dosage of Usnea extract powder is 200g / mu of water, once a day for 3 consecutive days. Then, randomly select 5 bass and observe the number of Trichodina parasites on the gills of the bass using the same method. At the same time, observe the behavior and feeding of the bass after medication to evaluate the therapeutic effect of Usnea extract powder on Trichodina disease in bass.

[0059] Results: On the second day of use, feeding speed improved significantly, competition for food was more pronounced, and the phenomenon of fish scattering and attacking each other disappeared. After three days of use, the average number of Trichodina parasites on the gill filaments of bass decreased from 40-50 per field of view to 0-1 per field of view (e.g., Figure 1 B) Simultaneously, the feeding speed of the bass on the feeding platform further increased, and the fish's feeding speed also significantly increased. This demonstrates that, according to Example 2, the Usnea extract powder, when applied throughout the pond, can have a good therapeutic effect on Trichodina disease in bass.

[0060] Test Example 3: The effect of Usnea Sulphuric Acid Powder (Example 3) on tapeworm disease in grass carp (worms).

[0061] Mr. Zhao, a farmer in Luoyang, Henan Province, had a 15-mu (approximately 1 hectare) grass carp pond with a stock of 10,000 kilograms of fish, each weighing around 3 liang (approximately 150 grams). Starting in early June, the fish began to eat less well than usual. On June 12th, 10 grass carp were randomly selected. Dissection and microscopic examination revealed a large number of tapeworms in their intestines, averaging over 30 tapeworms per fish. Figure 2 A) It almost filled the entire intestine of the grass carp.

[0062] Treatment method: The *Usnea sulforaphane* powder from Example 3 was mixed evenly with grass carp feed, and a binder was added to prepare a medicated feed with a *Usnea sulforaphane* powder content of 10 g / kg of feed. After feeding continuously for 5 days, 10 grass carp were randomly selected, and the number of tapeworms in their intestines was observed using the same method. Simultaneously, the behavioral characteristics and feeding behavior of the grass carp after medication were observed to evaluate the therapeutic effect of *Usnea sulforaphane* powder on grass carp tapeworm disease. Results: After 5 days of use, the feeding speed was significantly improved, and competition for food was obvious. Dissection of 10 randomly selected grass carp revealed that the number of tapeworms in their intestines had decreased to 0 (e.g., ...). Figure 2B) At the same time, the fish's feeding speed also increased significantly. This demonstrates that, according to Example 3, Usnea Sulphuric Acid Powder, when prepared as medicated feed and administered orally, can have a good therapeutic effect on tapeworm disease in grass carp.

[0063] Test Example 4: The effect of sodium urinate powder (Example 4) on crucian carp sporozoan disease (protozoa).

[0064] Sporozoania is one of the most serious diseases affecting crucian carp farming areas, typically occurring from April to October each year, with peak incidence in May and September. In the early stages of infection, white cysts appear on the gills, while the throat remains asymptomatic. In the middle stages, the gills become infected with increased mucus, and the gill filaments rot and swell. Infected fish often exhibit clustering and surfacing behavior downwind, and a small number die. In the later stages of infection, the throat swells or ulcerates, containing numerous sporozoania cysts, and mortality increases dramatically.

[0065] Experiment location and conditions: A crucian carp fry pond located in Nanping Town, Gong'an County, Jingzhou City, Hubei Province. The pond covers an area of ​​4.5 mu (approximately 0.23 hectares), with a size of about 300 fish per jin (approximately 0.5 kg) and a stock of 200,000 fish. Sampling observation and microscopic examination confirmed the fish to be in the late stage of crucian carp infection with pharyngeal cystis disease. A large number of crucian carp were dying daily, with more than 6,000 fish dying each day.

[0066] Treatment plan: Due to the severity of the case, the medication was administered both orally and topically, with an increased dosage.

[0067] External application: Apply 100g of sodium thiocyanate powder per acre of water to the entire pond once a day. When feeding, apply 50g to the feeding platform for 3 consecutive days.

[0068] For internal administration via feed mixing: Mix sodium usnic acid powder thoroughly with crucian carp feed, add binder to make medicated feed with sodium usnic acid powder content of 30g / kg of feed, and feed continuously for 5 days (medication cycle from June 26 to June 30).

[0069] The results are shown in Table 1: Before medication, more than 6,000 crucian carp died daily, and a large number of dead fish could be seen at the downwind end of the pond every day. Ten live crucian carp were randomly selected for necropsy; 7 fish had subcutaneous cysts, and 5 fish had cysts in the throat or gills (see Table 1). Figure 3 When crucian carp lesions are pressed into slides and examined under a microscope, a large number of sporozoites can be observed (see...). Figure 4 After using sodium thiocyanate powder, the mortality rate of crucian carp decreased rapidly. By the 5th day, the mortality rate had dropped to 30 fish, and almost no dead fish were visible downwind of the pond. Ten live crucian carp were randomly selected for necropsy. No sporangia were found under the skin, in the throat, or in the gills of any of the fish. Microscopic examination of the gill filaments revealed no sporozoan parasites (see...). Figure 5 ).

[0070] Table 1. Record of fish mortality and sampling observations before and after treatment during the use of sodium thiocyanate powder.

[0071]

[0072] Sporozoanosis is an extremely difficult disease to cure in crucian carp. Currently, there are no targeted treatments. When faced with this disease, most fishpond owners may have to abandon all the fish in the pond and carry out relevant pest control treatments before restarting the pond, which can easily cause a lot of losses. However, by using the sodium urate powder of this invention, through a combination of high-dose oral administration and external application, sporozoanosis in crucian carp can be effectively treated, with good clinical efficacy.

[0073] Test Example 5: The effect of sodium thiocyanate powder (Example 4) on Ichthyophthirius multifiliis disease (cilia) in carp.

[0074] In mid-September, a carp pond in Zhengzhou, Henan Province, with a total water volume of 10 mu (approximately 1.65 acres) and a stocking density of 3,000 fish per mu, began to show signs of poor feeding and scattering among the fish. More than 20 fish were dying every day. White spots were clearly visible on the dead or dying fish, raising suspicion of Ichthyophthirius multifiliis parasites. Five carp were randomly selected, and small amounts of gill filaments and caudal fins were cut off and pressed into slides. These slides were then examined under a microscope (10×10 magnification). It was found that a large number of Ichthyophthirius multifiliis parasites were found on the gills of the carp. The average number of Ichthyophthirius multifiliis was 18-20 per field of view.

[0075] Treatment method: Dissolve and dilute the sodium usnic acid powder of Example 4 in an appropriate amount of water, and then sprinkle it throughout the pond at a dosage of 200g / mu of water, once a day for 5 consecutive days. Then, randomly select 5 carp and observe the number of carp parasites using the same method. At the same time, observe the behavioral characteristics and feeding status of the carp after medication, and record the mortality rate to evaluate the therapeutic effect of sodium usnic acid powder on carp Ichthyophthirius multifiliis disease.

[0076] Results: On the third day of treatment, the number of carp deaths decreased from more than 20 per day before treatment to 3 per day. After five days of treatment, no more carp deaths occurred. After five days of treatment, feeding improved significantly, and the fish no longer exhibited scattering behavior. Sampling observation showed that the white spot symptoms on the carp's body surface disappeared. Microscopic examination of the gills revealed that the average number of Ichthyophthirius multifiliis parasites on the gill filaments decreased from 18-20 per field of view to 0-1 per field of view. This indicates that the sodium thiocyanate powder in Example 4, applied by whole-pond application, has a good therapeutic effect on Ichthyophthirius multifiliis disease in carp.

[0077] Test Example 6, Example 3: Effect of Usnea Sulphuric Acid Powder (Example 3) on Yellow Catfish Trichodina (Ciliates)

[0078] In Huzhou, Zhejiang Province, a yellow catfish farming pond with a total water volume of 8 mu (approximately 0.33 hectares) and a stocking density of 2,500 fish per mu (approximately 1,600 fish per hectare) experienced fish mortality in late May, with more than 50 fish dying daily. Upon sampling and observation, it was found that the diseased fish had increased mucus and ulceration on their body surface and gills, were emaciated, had reduced appetite, and sluggish swimming. They often floated downwind of the pond edge. Ten yellow catfish were randomly collected, and a small amount of gill filaments were cut from each fish, made into slides, and observed under a microscope (10×10x). It was found that the gills of the yellow catfish were infested with a large number of Chilodonella parasites. The number of Chilodonella was counted, with an average of more than 20 per field of view.

[0079] Treatment method: Dissolve and dilute the Usnea acid powder of Example 3 in an appropriate amount of water, and then sprinkle it throughout the pond at a dosage of 100g / mu of water, once a day for 5 consecutive days. Then, randomly select 10 yellow catfish and observe the number of parasites of Trichodina in the yellow catfish using the same method. At the same time, observe the behavioral characteristics and feeding of the yellow catfish after medication, and record the mortality rate to evaluate the therapeutic effect of Usnea acid powder on Trichodina disease in yellow catfish.

[0080] Results: On the third day of treatment, the number of yellow catfish deaths decreased from over 50 per day before treatment to 20 per day. After five days of treatment, the daily mortality rate dropped to below 5. Sampling observations showed a significant reduction in symptoms such as increased mucus and gill ulceration in the yellow catfish. Microscopic examination of the gills revealed that the average number of *Trichodina* parasites on the gill filaments decreased from over 20 per field of view to 0-2 per field of view. This demonstrates that the *Usnea oleifera* powder in Example 3, applied throughout the pond, has a good therapeutic effect on *Trichodina* disease in yellow catfish.

[0081] Test Example 7: Effect of Usnea Extract Powder (Example 2) on Cryptocaryon irritans (cilia) in Large Yellow Croaker

[0082] A case of Cryptocaryon irritans disease was discovered in fish farms of large yellow croaker in Ningde, Fujian. Affected fish developed numerous small white spots (approximately 1 mm in diameter) on their bodies, gills, and fins. Increased mucus production, punctate congestion on the epidermis at the infected sites, and pale gill filaments indicated anemia. The fish became sluggish, swimming in circles, exhibited lethargy, decreased appetite, and gradually became emaciated until death. Before medication, an average of more than 10 fish died per cage per day. Five large yellow croakers were randomly collected, and small amounts of gill filaments or white spots on the body surface were scraped off and pressed into slides. Microscopic examination (10×10x) revealed a large number of Cryptocaryon irritans parasites on the gills and body surface of the fish. The average number of parasites was >40 per field of view.

[0083] Treatment method: Dissolve and dilute the Usnea extract powder from Example 2 in an appropriate amount of water, and sprinkle 200g onto the fish during feeding. Simultaneously, mix the Usnea extract powder evenly with wild fish to make fish paste feed, and feed this to the fish at a dosage of 5g / kg of fish paste for 7 consecutive days. After 7 days of oral administration and fish spraying, randomly select 5 large yellow croakers and observe the number of Cryptocaryon irritans parasites on the fish using the same method. Simultaneously observe the behavioral characteristics and feeding behavior of the large yellow croakers after medication, and record mortality to evaluate the therapeutic effect of Usnea extract powder on Cryptocaryon irritans in large yellow croakers.

[0084] Results: After 7 days of medication, the mortality rate of large yellow croaker gradually decreased, from an average of more than 10 fish per net cage per day before medication to 1-2 fish per day, or even none at all. The vitality of the large yellow croaker significantly improved, and their feeding behavior was significantly enhanced. Sampling observation showed that the white spot symptoms on the body surface of the large yellow croaker disappeared. Microscopic examination of the gills and body surface mucus of the large yellow croaker revealed that the number of Ichthyophthirius multifiliis parasites on the fish body decreased from an average of >40 per field of view before medication to 0-3 per field of view. This indicates that the Usnea rubra extract powder in Example 2, when applied to the fish and administered orally in combination with medicated feed, can have a good therapeutic effect on Cryptocaryon irritans disease in large yellow croaker.

[0085] Test Example 8: The effect of Usnea extract powder (Example 2) on crustacean ciliate disease (cilia).

[0086] The water in the factory-farmed Litopenaeus vannamei (whiteleg shrimp) cultured in the Nanpai River of Huanghua, Hebei Province, was turbid and sticky. Ten shrimp were randomly selected for dissection and observation. The shrimp bodies, appendages, and gills were found to be heavily soiled. Microscopic examination of the appendages and gill filaments revealed a large number of worms attached to them. Due to the excessive number of worms, the shrimp's feeding was impaired, resulting in less intestinal food and lower appetite. The average number of worms was >30 per field of view.

[0087] Treatment method: Dissolve and dilute the Usnea extract powder from Example 2 in an appropriate amount of water, and then spray it throughout the pond at a dosage of 2g per cubic meter of water. Two hours after spraying, replace half of the water in the pond. Spray once every other day for a total of 3 times. At the same time, mix the Usnea extract powder evenly with the shrimp feed to make medicated feed, and feed it at a dosage of 10g / kg of feed, twice a day for 5 consecutive days. After the treatment ends, randomly select 10 shrimp and observe the number of diatoms attached to the appendages and gills of the shrimp using the same method, and observe the improvement in the shrimp's feeding after treatment.

[0088] Results: After the medication was administered, the shrimp's feeding improved significantly, the water turbidity was significantly reduced, and the water quality became clear. Upon sampling, the dirt attached to the shrimp's appendages and gills disappeared and the water returned to cleanliness. Microscopic examination revealed that the number of mitochondrial worms attached to the shrimp's appendages and gills decreased from an average of >30 per field of view before medication to an average of <2 per field of view. This indicates that the Usnea extract powder in Example 2, when applied by spraying and combined with medicated feed for oral administration, can have a good therapeutic effect on ciliate disease in shrimp.

[0089] Test Example 9: Effect of Usnea Sulphuric Acid Powder (Example 3) on Turbot Ciliate Disease (Ciliates)

[0090] A ciliate disease outbreak occurred in Mr. Qiu's turbot farm in Laizhou, Yantai, Shandong. Affected turbot exhibited increased mucus production and reddened, ulcerated lesions on their bodies and gills. The fish swam alone, became lethargic, and some lost their balance, exhibited decreased appetite, and darkened their color. Approximately 20 fish died per pond daily. Five turbot were randomly collected, and small amounts of gill filaments or mucus from the lesions were scraped and pressed into slides. Microscopic examination revealed a large number of shield-shaped ciliates parasitizing the gills and body surface of the turbot, with an average count of >10 filaments per field of view.

[0091] Treatment method: Dissolve and dilute the Usnea acid powder from Example 3 in an appropriate amount of water, and then sprinkle it throughout the pond at a rate of 5g per cubic meter of water. Simultaneously, mix the Usnea acid powder evenly with the feed to make medicated feed, and feed it at a dosage of 10g / kg of feed for 7 consecutive days. After 7 days of oral administration and pond-wide application of the medicated feed, randomly select 5 turbot and observe the number of parasitic scutellariae on the turbot using the same method. Simultaneously observe the behavioral characteristics and feeding behavior of the turbot after medication, and record mortality to evaluate the therapeutic effect of Usnea acid powder on turbot scutellariae.

[0092] Results: After 7 days of medication, the mortality rate of turbot gradually decreased, from an average of about 1020 fish per pond per day before medication to 1-2 fish per day, or even none at all. The turbot's vitality significantly improved, and their feeding behavior was noticeably better. Fish exhibiting behaviors such as swimming alone, sluggishness, and loss of balance were rarely seen. Sampling observation showed a reduction in mucus on the turbot's body surface and gills, and lesions gradually began to heal without further deterioration. Microscopic examination of the gills and body surface mucus revealed that the average number of parasitic scutociliates on the fish decreased from >10 per field of view to 0-1 per field of view. This demonstrates that Example 3, Usnea Sinensis Powder, administered orally in combination with prepared medicated feed, has a good therapeutic effect on scutociliate disease in turbot.

[0093] Multiple experimental results show that this invention, through administration methods such as veterinary drugs, feed additives, and functional feeds made from Usnea, Usnea extract, Usnea acid, and Usnea salts, has significant efficacy in eliminating or killing various parasites on the surface and inside the bodies of aquatic animals during the breeding process. These parasites include ciliates (Trichodina, Ichthyophthirius multifiliis, Chilodonella, Cryptocaryon irritans, crustacean ciliates, Shield ciliates, etc.), worms (Dactylogyrus, Tapeworms, etc.), and protozoa (Sporozoa, etc.). It can maintain the health of farmed animals, improve the success rate of breeding, achieve broad-spectrum and highly effective prevention and control of aquatic animal parasitic diseases, reduce the use of chemical drugs, and have good breeding, ecological, and social benefits. It is safe, broad-spectrum, and highly effective, and is conducive to large-scale promotion.

[0094] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. The application of Usnea powder, Usnea acid, or sodium usneate as the sole active ingredient in the preparation of drugs or feed additives or functional feeds for broad-spectrum antiparasitic treatments or treatments of parasitic infections in aquatic animals; wherein, The Usnea powder is Usnea pulverized material or Usnea extract; The parasite is one or more of the following: ciliates that infect fish or crustaceans, worms that infect fish, and protozoa that infect fish. The ciliates that infect fish or crustaceans are one or more of the following: Trichodina, Ichthyophthirius multifiliis, Chilodonella, Cryptocaryon irritans, Shield ciliates, and crustacean ciliates. The worms that infect fish are one or both of dactyloides and tapeworms; The protozoan that infects fish is *Carassius spp.* 2. The application according to claim 1, characterized in that, The crustacean ciliates mentioned are condensates.

3. The application according to claim 1, characterized in that, The drug is an oral medication or a medicated bath preparation.

4. The application according to claim 1, characterized in that, The drug comprises at least 0.2% by mass of usnic acid and / or usnic acid salt.

5. The application according to claim 1, characterized in that, The drug comprises at least 1% by mass of usnic acid and / or usnic acid salt.

6. The application according to claim 1, characterized in that, The drug comprises at least 2% by mass of usnic acid and / or usnic acid salt.

7. The application according to claim 1, characterized in that, The feed additive contains at least 0.002% by mass of isocyanate and / or isocyanate.

8. The application according to claim 1, characterized in that, The functional feed contains at least 0.0002% by mass of isoflavone and / or isoflavone salt.

9. The application according to any one of claims 1 to 8, characterized in that, Usnea extract is an organic solvent extract of Usnea.

10. The application according to claim 9, characterized in that, Organic solvents include one or more of acetone, chloroform, and glacial acetic acid.

11. The application according to any one of claims 1 to 8, characterized in that, The dosage form of the drug is any one of the following: powder, granule, solution, ointment, pourable, or injection.

12. The application according to any one of claims 1 to 8, characterized in that, The drug also contains pharmaceutically acceptable excipients.

13. The application according to any one of claims 1 to 8, characterized in that, Fish or crustacean aquatic animals include one or more of the following: grass carp, common carp, crucian carp, perch, yellow catfish, catfish, large yellow croaker, turbot, golden pomfret, tilapia, whiteleg shrimp, giant freshwater prawn, tiger prawn, Japanese prawn, Chinese shrimp, Chinese mitten crab, and mud crab.

14. The application according to claim 1 or 2, characterized in that, The parasites are one or more of the following: crucian carp dactylus, perch trichodina, grass carp tapeworm, crucian carp sporozoan, carp ichthyophthirius multifiliis, yellow catfish chilodonella, large yellow croaker cryptocaryon, Litopenaeus vannamei var. spp., and turbot scutellarinus.

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