Application of Chinese tulipa bark in preparation of product for preventing and / or treating marine fish pellioniasis

Abstract

The invention discloses an application of intermediary tulipa bark in preparation of a product for preventing and / or treating marine fish pellioniasis. The intaglio pomace can be an intaglio pomace preparation. The preparation comprises an active component, and the active component is poliodendron simsii, poliodendron simsii alcohol extract or poliodendron simsii water extract. An in-vitro insecticidal test and an animal infection test confirm that the pellionia obtusifolia can kill pellionia obtusifolia, the pellionia obtusifolia infection rate and the death rate of turbot pellionia obtusifolia can be obviously reduced, and the pellionia obtusifolia has a good effect of preventing and treating pellionia obtusifolia diseases. The Chinese tulip bark has the advantages of small toxicity to the turbot and environmental protection, and can reduce the environmental pollution caused by formaldehyde, copper sulfate and the like which are discharged in the turbot breeding process and are used for treating the shield ciliate disease.

Description

Technical Field

[0001] The invention belongs to the technical field of aquaculture disease prevention and control, and in particular relates to the application of Psoralea corylifolia in preparing products for preventing and / or treating marine fish clade ciliate disease. Background Art

[0002] Turbot (Scophthalmus maximus), also known as turbot, was originally naturally distributed in the coastal areas of Europe from the eastern Atlantic to the northeast. It lives on the seabed and leads a benthic life. Since its introduction into my country, turbot has created considerable economic benefits with its own advantages, and its artificial breeding has become one of the most important pillar industries in my country's marine aquaculture. However, with the continuous expansion of turbot farming scale and the continuous increase in breeding density, coupled with the slow progress in disease prevention and control, various breeding diseases have emerged one after another. Among the many breeding diseases, parasitic diseases caused by shield ciliates, cryptonuclei, wheelworms and flagellates have caused considerable losses to the turbot farming industry. The most serious of these diseases is shield ciliate disease, which is a systemic infectious disease that can quickly become fatal. Turbot diseases caused by shield ciliates are highly contagious and harmful. Especially for seedlings, once this disease breaks out, farmers often suffer huge economic losses. The main pathogen is Heterobranchus crabii, which can enter the aquaculture system through a variety of different pathways and cause disease in turbot. Shield ciliates are not only found in large numbers in the diseased parts, body surface and gills of turbot, but also in large numbers in organs such as the liver, spleen, kidneys, brain, heart, and ascites. In the early stages of infection, white spots will appear on the body surface of the diseased fish, and mucus secretion will increase; as the disease progresses, the tissue at the lesion will become red and swollen, and it will feel soft and swollen when touched; when the disease is serious, lesions will also appear on the skin of the fish's trunk, and even develop to the point of ulceration and bleeding. Sick turbot usually have a darker body color, significantly reduced vitality, reduced food intake, and slower growth rate; they no longer cluster in the aquaculture pond, but disperse, and often show abnormal behavior of swimming in circles. Moreover, ciliates can infect most internal organs, including brain tissue.

[0003] At present, there are not many effective treatments for shield ciliate diseases in China. In the actual breeding production process, in order to prevent and control the occurrence of diseases, antibiotics and chemical disinfectants are often used to treat fish diseases. Therefore, the search for new, safe, reliable and environmentally friendly insecticides has become an urgent problem that needs to be solved.

[0004] Aopio bark belongs to the genus Liriodendron of the Magnoliaceae family, also known as horse laurel bark. It is warm in nature and spicy in taste, and has the effects of dispelling wind and dampness, dispersing cold and relieving cough. It is generally used to treat rheumatic arthralgia, wind-cold cough and other diseases. There is no report that Aopio bark can kill shield ciliates. Summary of the invention

[0005] The purpose of the present invention is to provide an application of the cortex ophiopogonis in the preparation of a product for preventing and / or treating scutellaria disease in marine fish. The present invention has the advantages of low toxicity to turbot and environmental protection, and can reduce the pollution to the environment caused by formaldehyde, copper sulfate, etc. discharged during the turbot breeding process due to the treatment of scutellaria disease.

[0006] In a first aspect, the present invention provides the use of Psoralea corylifolia or its preparation in the preparation of a product for preventing and / or treating chelonian disease in marine fish.

[0007] In at least one embodiment of the present invention, the marine fish is turbot.

[0008] In at least one embodiment of the present invention, the Phellodendron chinense or its preparation is used as the only active ingredient; The preparation includes an active ingredient, which is the cortex phellodendri, an alcohol extract of the cortex phellodendri or an aqueous extract of the cortex phellodendri. It is understandable that in order to facilitate the preparation of various dosage forms, the preparation may also include excipients acceptable in traditional Chinese medicine preparations.

[0009] In at least one embodiment of the present invention, the product is a medicine or feed. According to its mode of administration (such as oral administration or medicinal bath), the dosage form of the preparation includes but is not limited to tablets, powders, granules, decoctions, wine preparations, etc.

[0010] As an example, further, the preparation is Aopikopi powder, and its preparation method comprises the following steps: taking Aopikopi medicinal material, crushing it, and passing it through a 100-mesh sieve. Optionally, its usage and dosage are as follows: adding 100 to 1000 g (such as 200 g) of Aopikopi powder to every 100 kg of feed, and feeding the diseased turbot.

[0011] In an alternative embodiment, the preparation is an alcohol extract of conophytum corydalis; the preparation method of the alcohol extract of conophytum corydalis comprises the following steps: taking conophytum corydalis, adding ethanol for immersion extraction. Further, the ratio of conophytum corydalis to ethanol is 1000g: (8000-12000) mL, including but not limited to 1000g: 10000mL; the mass concentration of ethanol is 95%-100%, including but not limited to food grade 95% ethanol (aqueous solution); it can be understood that 100% is anhydrous ethanol; in the immersion extraction step, the temperature is 25°C-50°C, including but not limited to room temperature 25°C; in the immersion extraction step, each extraction is 48-96 hours, including but not limited to 48 hours; in the immersion extraction step, the number of extractions is 2-3 times, and the extracts are combined. Further, after the extraction, the extract is also concentrated to 1g / ml in terms of crude drug amount. Optionally, the usage and dosage are as follows: take 1000g of conophaga bark, extract it with ethanol, and then sprinkle the extract into 2000kg of water (breeding density 30-40 fish / m³) to perform a medicated bath on the diseased turbot.

[0012] In an alternative embodiment, the preparation method of the water extract of the concave poplar bark comprises the following steps: the preparation method of the water extract of the concave poplar bark comprises the following steps: taking the concave poplar bark, adding water for decoction, and concentrating after decoction. Further, the ratio of the concave poplar bark to water is 1000g: (8000-12000mL), including but not limited to 1000g: 10000mL; in the decoction step, decoction is performed 2-3 times (such as 2 times), each time for 1 hour; the concentration is concentrated to 1g / ml in terms of crude drug amount.

[0013] In the above application of the present invention, the therapeutic effect is specifically reflected in reducing the number of parasitic ciliates of turbot and / or reducing the mortality rate of turbot.

[0014] In a second aspect, the present invention provides a marine fish feed, comprising a basic fish feed and a feed additive, wherein the feed additive is Psoralea corylifolia or a preparation thereof.

[0015] In at least one embodiment of the present invention, the marine fish is turbot. As an example, the composition of the basic fish feed is as follows, in terms of mass percentage: 45% to 50% fish meal, 20% to 25% soybean meal, 15% to 20% wheat flour, 5% to 8% fish oil, 2% to 3% soybean lecithin, 1% to 2% calcium dihydrogen phosphate, 1% to 1.5% vitamin premix, and 1% to 1.5% mineral premix. It can be reasonably adjusted according to the needs of turbot at different growth stages.

[0016] In at least one embodiment of the present invention, the Phellodendron chinense or its preparation is used as the only active ingredient; The preparation includes an active ingredient, which is Conococortex opposita, Conococortex opposita alcohol extract or Conococortex opposita water extract; it is understandable that in order to facilitate the preparation of various dosage forms, the preparation may also include excipients acceptable in each preparation; the dosage form of the preparation may be reasonably selected according to the type of feed.

[0017] Calculated on the basis of Psoralea corylifolia, the mass percentage of the feed additive in the marine fish feed is 0.1% to 1%. As an example, the preparation is Psoralea corylifolia powder, and the mass percentage of Psoralea corylifolia powder in the marine fish feed is 0.2%.

[0018] In a third aspect, the present invention provides the use of Psoralea corylifolia or its preparation in any of the following: A1) Use in the preparation of products for killing shield ciliates; A2) Killing of shield ciliates in vitro.

[0019] In the above application, the in vitro killing of shield ciliates is for non-therapeutic purposes and does not include wounds and infected areas.

[0020] In at least one embodiment of the present invention, the Cortex Phellodendri or its preparation is used as the sole active ingredient.

[0021] The preparation includes an active ingredient, and the active ingredient is conophytum corydalis, conophytum cory ethanol extract or conophytum cory water extract; it is understandable that in order to facilitate the preparation of various dosage forms, the preparation may also include excipients acceptable in each preparation. The dosage form of the preparation can be reasonably selected according to its application scenario. Optionally, the product is an insecticide.

[0022] In a preferred embodiment of the present invention, the active ingredient is an alcohol extract of conophytum corydalis; the preparation method of the alcohol extract of conophytum corydalis comprises the following steps: taking conophytum corydalis, adding ethanol for immersion extraction. Furthermore, the ratio of conophytum corydalis to ethanol is 1000g: (8000-12000) mL, including but not limited to 1000g: 10000mL; the mass concentration of ethanol is 95%-100%, including but not limited to food grade 95% ethanol; it can be understood that 100% is anhydrous ethanol; in the immersion extraction step, the temperature is 25°C-50°C, including but not limited to room temperature 25°C; in the immersion extraction step, each extraction is 48-96 hours, including but not limited to 48 hours; in the immersion extraction step, the number of extractions is 2-3 times, and the extracts are combined. Furthermore, after the extraction, the extract is also concentrated to 1g / ml in terms of crude drug amount.

[0023] In an alternative embodiment, the preparation method of the water extract of the concave poplar bark comprises the following steps: the preparation method of the water extract of the concave poplar bark comprises the following steps: taking the concave poplar bark, adding water for decoction, and concentrating after decoction. Further, the ratio of the concave poplar bark to water is 1000g: (8000-12000mL), including but not limited to 1000g: 10000mL; in the decoction step, decoction is performed 2-3 times (such as 2 times), each time for 1 hour; the concentration is concentrated to 1g / mL in terms of crude drug amount.

[0024] Optionally, based on the Conopsis cortex, the concentration (based on the amount of crude drug) of the Conopsis cortex or its preparation added to the water body (such as seawater) infected with Shield Ciliate is ≥0.25 g / L, such as 0.25 g / L.

[0025] The present invention has the following beneficial effects: The invention verifies through in vitro insecticidal tests and animal infection tests that the addition of Psoralea corylifolia can kill shield ciliates, can significantly reduce the infection rate and mortality rate of shield ciliates of turbot, and has a good effect of preventing and treating shield ciliates diseases.

[0026] The present invention discovers for the first time that the cortex pectinata has the effect of killing shield ciliates, can significantly reduce the mortality rate of turbot shield ciliate disease, and reduces the parasitic number of turbot shield ciliates. It has the advantages of low toxicity to turbot and environmental protection, and can reduce the pollution to the environment caused by formaldehyde, copper sulfate and the like emitted during the turbot breeding process due to the treatment of shield ciliate disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 This is a photo of the diseased turbot temporarily raised in Example 1 of the present invention.

[0028] Figure 2 These are photos of the grouping experiment in Example 1 of the present invention. DETAILED DESCRIPTION

[0029] The present invention is further described in detail below in conjunction with specific embodiments, and the examples provided are only for illustrating the present invention, rather than for limiting the scope of the present invention. The examples provided below can be used as a guide for further improvements by those of ordinary skill in the art, and do not constitute a limitation of the present invention in any way.

[0030] The methods used in the following examples, unless otherwise specified, are all conventional methods, carried out according to the techniques or conditions described in the literature in the art or according to the product instructions. The materials, reagents, etc. used in the following examples, unless otherwise specified, can all be obtained from commercial sources.

[0031] The Aopika bark medicinal material in the following embodiments is a Chinese herbal medicine purchased from Chengdu Yuhetang E-Commerce Co., Ltd.

[0032] Preparation Example 1: Preparation of Aopuki Powder Take the Aopikopi medicinal material, grind it, and pass it through a 100-mesh sieve to obtain the Aopikopi powder.

[0033] Dosage and Administration: Take 0.2 kg of the above powder, mix it into 100 kg of feed, and feed according to the daily intake.

[0034] Preparation Example 2: Preparation of alcohol extract of Psoralea corylifolia Take 1000 g of the cortex of Psoralea corylifolia, add 10000 mL of food-grade 95% ethanol, immerse and extract for 48 hours, extract twice, combine the extracts, and concentrate under reduced pressure to 1000 mL.

[0035] Dosage and administration: Take 1L of the above extract and sprinkle it into 2000kg of seawater (breeding density 30-40 fish / m³) to perform a medicated bath on turbot infected with shield ciliate disease for 30 minutes a day for 5-7 consecutive days.

[0036] Preparation Example 3: Preparation of Aqueous Extract of Psoralea corylifolia Take 1000g of Phellodendron chinense bark, add 10000mL of water, decoct twice, each time for 1 hour, combine the decoctions, and concentrate under reduced pressure to 1000mL.

[0037] Example 1, Preparation Example 1 Pharmacodynamic Experiment This example is used to observe the efficacy of Preparation Example 1 in preventing and treating turbot shield ciliate disease.

[0038] The following is an explanation of the anti-shield ciliate test of Aopikovia divaricata with reference to specific experiments.

[0039] A turbot farm with shield ciliates disease was selected. The infected turbot were confirmed to have shield ciliates by microscopic observation of gills. The infected turbot were selected and transferred to a temporary storage pond ( Figure 1 ), 30 turbot (200g-300g / piece) infected with shield ciliate disease were selected and randomly divided into 2 groups ( Figure 2 ), after grouping and acclimatization, one group was administered with the feed mixed with Preparation Example 1. 0.2 kg of Preparation Example 1 was taken and mixed into 100 kg of feed (formula see Table 1), and fed at 3% of the feeding amount. The other group was used as the model control group, and no drugs were given. The rearing temperature was 18°C. During the experiment, the diseased gills of the test group and the model control group of Preparation Example 1 were gently scraped with an inoculation loop, smeared on a slide, and the activity and number of ciliates were observed under a microscope. The experiment was carried out for 5 days, and the results are shown in Table 2.

[0040]

[0041] Evaluation criteria: A lot of them: Under the 100x lens, there are few spaces between the insects, and they are almost close together, with the entire field of view filled with insects. The score is 6 points.

[0042] Many: More than 20 insects in the field of view at 100x magnification. Score 5 points.

[0043] Medium: No more than 20 insects can be seen in the field of view at 100x magnification. Score: 4 points.

[0044] Few: Under 100x lens, there are no more than 10 insects in the field of view. Score 3 points.

[0045] Very few: No more than 3 insects in the field of view at 100x magnification. Score 1 point.

[0046] No: No parasites in the field of view under 100x lens, and no parasites after changing the field of view. The score is 0.

[0047]

[0048] "*" indicates that the evaluation score of the Example group after 5 days of medication was significantly different from that of the model group.

[0049] The experimental results showed that Aopikopi can significantly reduce the number of parasitic shield ciliates of turbot and reduce the mortality rate, while the number of parasitic shield ciliates in the model group is still increasing. Aopikopi has a significant effect compared with the model control group, indicating that Aopikopi has a significant effect in resisting the shield ciliate disease of turbot.

[0050] Pharmacological efficacy experiments of Example 2, Preparation Example 2 and Preparation Example 3 In vitro insecticidal test: Take a 96-well plate, first add 0.6 mL of seawater to each well, then add 0.6 mL of Preparation Example 2 to the first well, dilute by multiples, and dilute to the 12th well. The drug concentrations are 1 / 2-1 / 4096 (for example, 0.6 ml of drug solution + 0.6 ml of seawater corresponds to a drug concentration of 1:2, and then dilute once by a multiple of 2 to a drug concentration of 1:4); according to this method, dilute Preparation Example 3 and food-grade 95% ethanol, take 120 μL of each diluted sample, add it to a 96-well cell culture plate, and then add 120 μL of shield ciliate solution to each well (the shield ciliates are derived from clinically sick turbot, cultured and propagated in vitro, and can be used as an in vitro insecticidal test when there are more than 10 shield ciliates in the 100x microscope field of view), gently shake the cell culture plate for 1 minute, then let it stand for 2 hours, and observe whether the ciliates are dead under an inverted microscope to determine the drug effect. At the same time, a blank control group was set up. The concentration of shield ciliates in the blank control group was the same as that in the experimental group, and only an equal amount of seawater or food-grade 95% ethanol was added.

[0051] The minimum insecticidal concentration was the lowest concentration at which all the shield ciliates died under microscopic examination. The results showed that the minimum insecticidal concentration of the 95% food-grade ethanol solvent control group was 1:8, the in vitro minimum insecticidal concentration of Preparation Example 2 was 1:4096, and the minimum insecticidal concentration of Preparation Example 3 was 1:512 (0.000244 ml / ml). It can be seen that the alcohol extract of the concave pectin has a good effect of killing shield ciliates, and the water extract of the concave pectin also has an in vitro effect of killing shield ciliates.

[0052] The present invention has been described in detail above. For those skilled in the art, without departing from the purpose and scope of the present invention, can implement the present invention in a wider range under equivalent parameters, concentrations and conditions. Although the present invention provides specific embodiments, it should be understood that the present invention can be further improved. In a word, according to the principle of the present invention, the application is intended to include any changes, uses or improvements to the present invention, including departing from the disclosed scope in the application and the changes made with conventional techniques known in the art.

Claims

1. Use of the Cortex Phellodendri or its preparation in the preparation of products for preventing and / or treating marine fish Shield ciliate disease.

2. The use according to claim 1, characterized in that: The marine fish is turbot.

3. The use according to claim 1 or 2, characterized in that: The preparation comprises an active ingredient, and the active ingredient is the cortex ophiopogonis, an alcohol extract of the cortex ophiopogonis or an aqueous extract of the cortex ophiopogonis.

4. The use according to claim 1 or 2, characterized in that: The product is a medicine or a feed.

5. A marine fish feed, characterized in that: The invention comprises a basic fish feed and a feed additive, wherein the feed additive is Psoralea corylifolia or a preparation thereof.

6. The marine fish feed according to claim 5, characterized in that: The marine fish is turbot; The preparation is Aopurus cortex powder, Aopurus cortex alcohol extract or Aopurus cortex water extract; Calculated on the basis of conoporin, the mass percentage of the feed additive in the marine fish feed is 0.1% to 1%.

7. Use of Aopak bark or its preparation in any of the following: A1) Preparation of products for killing shield ciliates; A2) Killing of shield ciliates in vitro.

8. The use according to claim 7, characterized in that: The preparation comprises an active ingredient, and the active ingredient is the cortex ophiopogonis, an alcohol extract of the cortex ophiopogonis or an aqueous extract of the cortex ophiopogonis.

9. The use according to claim 8, characterized in that: The method for preparing the alcohol extract of the conocarp bark comprises the following steps: taking the conocarp bark and adding ethanol for immersion extraction.

10. The use according to claim 9, characterized in that: The ratio of the conocarp to ethanol is 1000 g: (8000-12000) mL; The mass concentration of the ethanol is 95% to 100%; In the soaking and extraction step, the temperature is 25°C to 50°C; In the maceration extraction step, each extraction takes 48 to 96 hours; In the immersion extraction step, the extraction times are 2 to 3 times, and the extracts are combined.

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