Use of extract of cinnamomum osmophloeum kanehira

Abstract

One use of cinnamon leaf extract is for preparing a drug to enhance the immunity of shrimp. The cinnamon leaf extract is administered to a shrimp to enhance its immunity. The preparation method of the cinnamon leaf extract includes: extracting a cinnamon leaf sample at a temperature of 90-100°C using water as an extraction solvent for 10-20 minutes to obtain a crude cinnamon leaf extract; and mixing the crude cinnamon leaf extract with 3-5 times its volume of an aqueous ethanol solution to precipitate the cinnamon polysaccharides in the crude cinnamon leaf extract to obtain the cinnamon leaf extract.

Description

Technical Field

[0001] This invention relates to the use of a cinnamon leaf extract, and more particularly to the use of the cinnamon leaf extract in the preparation of a drug to enhance the immunity of shrimp. Prior Technology

[0002] In recent years, shrimp farming in Taiwan has flourished. Shrimp not only serve as a food source but also generate substantial economic benefits for the country through exports. Due to the rapid development of shrimp farming, coupled with high land prices, farmers are forced to adopt high-density farming to reduce costs.

[0003] High-density farming can lead to a deterioration of the farming environment, making it easier for shrimp to spread diseases and thus increasing the mortality rate. In order to reduce the mortality rate, farmers often add antibiotics to the feed. However, the overuse of antibiotics not only causes allergies in consumers who eat shrimp, but may also induce antibiotic resistance in pathogens. Therefore, further improvements are still needed. Summary of the Invention

[0004] The purpose of this invention is to provide a use for cinnamon leaf extract, which can enhance the immunity of shrimp, thus eliminating the need for antibiotic users.

[0005] The purpose of the cinnamon leaf extract of this invention is to prepare a drug that enhances the immunity of shrimp. The cinnamon leaf extract is administered to a shrimp to enhance its immunity. The preparation method of the cinnamon leaf extract includes: extracting a cinnamon leaf sample at a temperature of 90-100°C using water as an extraction solvent for 10-20 minutes to obtain a crude cinnamon leaf extract; and mixing the crude cinnamon leaf extract with 3-5 times its volume of an aqueous ethanol solution to precipitate the polysaccharides in the crude cinnamon leaf extract to obtain the cinnamon leaf extract. For example, the crude cinnamon leaf extract can be obtained by extracting the cinnamon leaf sample at a temperature of 95°C for 15 minutes, and the crude cinnamon leaf extract can then be mixed with 3 times its volume of an aqueous ethanol solution to precipitate the polysaccharides in the crude cinnamon leaf extract.

[0006] The use of the cinnamon leaf extract of this invention is that the active ingredients extracted from cinnamon leaf samples can promote the phagocytic rate of shrimp hemoglobin cells, thereby reducing the mortality of shrimp caused by pathogen attacks. This can reduce the amount of antibiotics used by aquaculture farmers during the aquaculture process, which is the effect of this invention.

[0007] The use of cinnamon leaf extract of the present invention, wherein the ethanol aqueous solution can be an ethanol aqueous solution with an ethanol concentration of 90-99.9%; for example, the ethanol aqueous solution can be an ethanol aqueous solution with an ethanol concentration of 95%.

[0008] The use of the cinnamon leaf extract of the present invention, wherein the polysaccharide system in the crude cinnamon leaf extract can precipitate at a temperature of 4-8°C for 16-24 hours; for example, the polysaccharide system in the crude cinnamon leaf extract can precipitate at a temperature of 4°C for 16 hours.

[0009] The use of the cinnamon leaf extract of the present invention includes the injection of the cinnamon leaf extract into the shrimp body; for example, the cinnamon leaf extract can be injected into the ventral sinusoids of the cephalothorax of the shrimp body. Thereby, the cinnamon leaf extract can flow to various tissues via the hemolymphatic system of the shrimp body, thereby effectively enhancing the shrimp's immune capacity.

[0010] The present invention relates to the use of cinnamon leaf extract, wherein the cinnamon leaf extract can be injected into the shrimp at a dose of 0.375 μg per gram of shrimp body weight. This allows for a better enhancement of the shrimp's immune function through the dosage of the cinnamon leaf extract. Simple Explanation of the Diagram

[0011] [Figure 1] Phagocytic activity of hemoglobin cells in freshwater long-armed shrimp at different time points in Experiment (A). [Figure 2] Survival rate of freshwater long-armed shrimp in each group at different time points in experiment (B). Implementation

[0012] To make the above and other objects, features and advantages of the present invention more apparent and understandable, preferred embodiments of the present invention are described below in detail with reference to the accompanying drawings:

[0013] The term "shrimp" as used in this invention can refer to farmed shrimp, including but not limited to: white shrimp (Litopenaeus vannamei), grass shrimp (Penaeus monodon), tiger prawn (Marsupenaeus japonicus), Chinese grass shrimp (Fenneropenaeus chinensis), Indian shrimp (Fenneropenaeus indicus), sand shrimp (Metapenaeus ensis barbata), red-tailed shrimp (Penaeus penicillatus), and freshwater long-armed shrimp (Macrobrachium rosenbergii), etc.

[0014] The cinnamon leaf extract of this invention can refer to any extract obtained from a cinnamon leaf sample. In this embodiment, the cinnamon leaf extract is obtained by extracting a cinnamon leaf sample with an extractant. The cinnamon leaf sample can be a leaf sample of *Cinnamomum osmophloeum* Kanehira (also known as Taiwan native cinnamon or false cinnamon), belonging to the genus *Cinnamomum* of the family Lauaceae. For example, the cinnamon leaf extract can be prepared by the preparation method described below, which may include: a sample providing step, an extraction step, and a precipitation step.

[0015] In the sample provision step, the operator can directly perform the extraction step after obtaining the cinnamon leaf sample, or the cinnamon leaf sample can be pre-dried before the extraction step, for example, reducing the moisture content of the cinnamon leaf sample to 3-15%. In this embodiment, the cinnamon leaf sample (moisture content of approximately 42%) is placed in an oven at 50°C and dried for 24 hours to obtain a dried cinnamon leaf sample with a moisture content of approximately 8%. Furthermore, the cinnamon leaf sample can also be pre-crushed (for example, to a particle size ≤0.18 mm) to increase the contact surface area of ​​the extractant used in the extraction step, thereby improving the subsequent extraction efficiency.

[0016] In this extraction step, the operator can choose water as the extractant (e.g., distilled water, deionized water, reverse osmosis water, and ultrapure water); however, the operator can also choose other extractants with similar polarity according to the application requirements to extract the active substances in the cinnamon leaf sample that inhibit the formation of advanced glycation end products, thereby obtaining the crude cinnamon leaf extract. Furthermore, the operator can adjust the temperature and / or time of extraction with the extractant, for example, adjusting it to a temperature between 90 and 100°C and / or between 10 and 20 minutes. The aforementioned extraction can also be repeated several times to ensure that the active ingredients rich in the cinnamon leaf sample are completely released into the extractant. This is widely used by those skilled in the art and will not be elaborated upon here. In this embodiment, the operator chooses deionized water as the extractant and extracts at 95°C for 15 minutes to obtain a crude cinnamon leaf extract.

[0017] Furthermore, before extracting the cinnamon leaf sample with the extractant, the operator preferably adjusts the ratio of the cinnamon leaf sample to the extractant to ensure that the active ingredients in the cinnamon leaf sample are effectively dissolved in the extractant with the minimum amount of extractant, thereby reducing the preparation cost of the cinnamon leaf extract. For example, the operator can mix the cinnamon leaf sample and the extractant at a ratio between 500 and 2,000 ml per 100 grams of cinnamon leaf sample; in this embodiment, the operator mixes 30 grams of cinnamon leaf sample with 400 ml of extract.

[0018] In this precipitation step, the worker can mix the crude cinnamon leaf extract with a precipitating agent to precipitate the cinnamon polysaccharides in the crude cinnamon leaf extract, thereby obtaining the cinnamon leaf extract. For example, the precipitating agent can be an aqueous ethanol solution (e.g., an aqueous ethanol solution with an ethanol concentration of 90-99.9%). The worker mixes the crude cinnamon leaf extract with 3-5 times its volume of the aqueous ethanol solution and allows the mixture to stand at 4-8°C for 16-24 hours to precipitate the cinnamon polysaccharides. In this embodiment, the worker mixes the crude cinnamon leaf extract with 3 times its volume of the aqueous ethanol solution (ethanol concentration of 95%) and allows the mixture to stand at 4°C for 16 hours to precipitate the cinnamon polysaccharides. Finally, the mixture is dried at 50°C for 6 hours to obtain the cinnamon leaf extract.

[0019] Cinnamon leaf extract can increase the phagocytic rate of shrimp hemoglobin cells and reduce shrimp mortality caused by pathogen attacks. Therefore, an effective dose of cinnamon leaf extract can be administered to the shrimp to enhance their immunity.

[0020] For example, the cinnamon leaf extract can be administered to the shrimp via injection. For instance, the cinnamon leaf extract can be injected into the ventral sinusoids of the cephalothorax of the shrimp. These ventral sinusoids are close to the heart and are where the shrimp's hemolymph is concentrated, so the cinnamon leaf extract can flow to various tissues via the shrimp's hemolymph. In this embodiment, a shrimp weighing approximately 8 grams is selected, and the cinnamon leaf extract is administered to the shrimp at a dose of 0.375 μg per gram of shrimp body weight.

[0021] To verify that the cinnamon leaf extract of this invention does indeed have the activity of enhancing the immune function of shrimp, the following experiments were conducted using freshwater long-armed shrimp:

[0022] (A) Efficacy against Lactococcus gasseri

[0023] Please refer to Table 1. In this experiment, different doses of cinnamon leaf extract were injected into freshwater giant pandas. One day after the injection of cinnamon leaf extract, each group of freshwater giant pandas was injected with Lactococcus garvieae to infect them with Lactococcus garvieae (infectious dose of 2 × 10⁷ CFU / shrimp body). One hour later, hemolymph was collected, and the phagocytic activity of hemolymph cells in each group of freshwater giant pandas was further tested to evaluate the efficacy against Lactococcus garvieae.

[0024] Table 1. Dosage of cinnamon leaf extract in each group of this experiment Group Cinnamon leaf extract A0 0 A1 3 μg / shrimp body

[0025] Referring to Figure 1, compared to Group A0 freshwater long-armed shrimp that were not given the cinnamon leaf extract of the present invention, the phagocytic activity of hemoglobin cells in Group A1 freshwater long-armed shrimp given 3 μg of the cinnamon leaf extract of the present invention was improved, indicating that the cinnamon leaf extract of the present invention has the activity of enhancing the clearance of bacteria in hemolymph.

[0026] (B) Survival rate after Lactococcus gasseri infection

[0027] Next, referring to Table 2, this experiment also injected different doses of cinnamon leaf extract into the freshwater giant panda. Two days after the injection of cinnamon leaf extract, each group of freshwater giant pandas was injected with Lactococcus gasseri to infect the freshwater giant pandas with Lactococcus gasseri (infectious dose of 2 x 10⁵ CFU / shrimp body). The mortality rate of each group of freshwater giant pandas was calculated at 0, 24, 48, 72, 96, 120, 168 and 168 hours after infection.

[0028] Table 2. Dosage of cinnamon leaf extract in each group of this experiment Group Lactococcus gasseri Cinnamon leaf extract B0 X 0 B1 O 0 B2 O 3 μg / shrimp body

[0029] Referring to Figure 2, compared to Group B1 freshwater prawns that were not given the cinnamon leaf extract of this invention, Group B2 freshwater prawns given 3 μg of the cinnamon leaf extract of this invention showed a significant reduction in mortality within 96 to 168 hours. This indicates that the cinnamon leaf extract of this invention can effectively prevent infection by Lactococcus gasseri and reduce the risk of death of freshwater prawns due to Lactococcus gasseri infection.

[0030] In summary, the use of the cinnamon leaf extract of the present invention is that the active ingredients extracted from cinnamon leaf samples can promote the phagocytic rate of shrimp hemoglobin cells and the ability to clear bacteria from hemolymph, and reduce the mortality of shrimp caused by pathogen attacks. Therefore, it can reduce the amount of antibiotics used by aquaculture farmers in the aquaculture process, which is the effect of the present invention.

[0031] It is particularly important to note that freshwater prawns infected with Lactococcus gasseri not only exhibit symptoms such as slow growth, loss of appetite, and abnormal behavior, but also develop lesions such as muscle tissue bleaching and hepatopancreatic enlargement. Furthermore, the mortality rate of freshwater prawns infected with Lactococcus gasseri is extremely high, often causing significant losses to aquaculture farmers. Therefore, preventing and controlling Lactococcus gasseri infection is crucial for ensuring the stability and sustainable development of the aquaculture industry.

[0032] Although the present invention has been disclosed using the above preferred embodiments, it is not intended to limit the present invention. Any modifications and alterations made by those skilled in the art to the above embodiments without departing from the spirit and scope of the present invention shall still fall within the scope of the technology protected by the present invention. Therefore, the scope of protection of the present invention shall be determined by the appended claims.

[0033] none

Claims

1. A method for farming freshwater giant pandas to prevent death from Lactococcus gasseri infection, comprising: injecting a cinnamon leaf extract into the ventral sinusoids of the cephalothorax of a freshwater giant panda at a dose of 0.375 μg / g of the panda's body weight, thereby preventing death from Lactococcus gasseri infection; wherein, The method for preparing the cinnamon leaf extract includes: extracting a cinnamon leaf sample for 10-20 minutes at a temperature of 90-100°C using water as an extraction solvent to obtain a crude cinnamon leaf extract; and mixing the crude cinnamon leaf extract with 3-5 times its volume of an aqueous ethanol solution to precipitate the cinnamon polysaccharides in the crude cinnamon leaf extract to obtain the cinnamon leaf extract.

2. The farming method for preventing the death of freshwater giant prawns due to Lactococcus gasseri infection, as requested in Item 1, wherein... The crude extract of cinnamon leaves was obtained by extracting the cinnamon leaf sample at 95°C for 15 minutes.

3. The farming method for preventing the death of freshwater giant prawns due to Lactococcus gasseri infection, as requested in Item 1, wherein... The crude cinnamon leaf extract is prepared by mixing three times its volume of an aqueous ethanol solution to precipitate the cinnamon polysaccharides from the crude cinnamon extract.

4. The farming method for preventing the death of freshwater giant prawns due to Lactococcus gasseri infection, as requested in Item 1, wherein... The ethanol-water solution is an ethanol-water solution with an ethanol concentration of 90-99.9%.

5. The farming method for preventing the death of freshwater giant prawns due to Lactococcus gasseri infection, as requested in item 4, wherein... This ethanol-water solution is an ethanol-water solution with an ethanol concentration of 95%.

6. The farming method for preventing the death of freshwater giant prawns due to Lactococcus gasseri infection, as requested in Item 1, wherein... The cinnamon polysaccharides in the crude cinnamon leaf extract precipitate out at a temperature of 4–8°C for 16–24 hours.

7. The farming method for preventing the death of freshwater giant prawns due to Lactococcus gasseri infection, as requested in item 6, wherein... The cinnamon polysaccharides in the crude cinnamon leaf extract precipitated at 4°C for 16 hours.

Images