Avian influenza virus poisoning agent, virus killing application and virus killing method

Abstract

The invention discloses an avian influenza virus poisoning agent, virus killing application and a virus killing method. And directly using the sodium chloride aerosol with the average particle size of 800nm and the particle size distribution of 500nm-8mu m as the avian influenza virus poisoning agent. The sodium chloride aerosol has the advantages that the sodium chloride aerosol with the average particle size of 800nm and the particle size distribution of 100nm-8mu m is prepared by combining an anti-solvent, a spray drying method and specific process parameters and is directly used as the avian influenza virus poisoning agent, the sodium chloride aerosol has the virus killing effect in a specific dosage and time range, bacterial cells can be dehydrated by a high-concentration sodium chloride solution through osmotic pressure difference, and the concentration of the sodium chloride solution in the avian influenza virus poisoning agent is reduced. The sodium chloride aerosol is used for killing the H9N2 subtype avian influenza virus, the cell membrane structure of the H9N2 subtype avian influenza virus is damaged, the growth inhibition or death of the bacteria is caused, especially, 100% killing of the H9N2 subtype avian influenza virus can be achieved with a small amount of sodium chloride aerosol and a short time, and the effect is very

Description

Technical Field

[0001] This invention relates to a method for killing avian influenza virus (H9N2 subtype), specifically an avian influenza virus disinfectant, its use in killing the virus, and a method for killing the virus. Background Technology

[0002] Highly pathogenic avian influenza, as well as animal diseases such as Newcastle disease, mad cow disease, and African swine fever, have caused enormous economic losses to the livestock industry. Among these, zoonotic diseases also threaten human life. Currently, there are no long-lasting vaccines for some livestock viruses. Disinfection of farms, primarily using disinfectants, remains the most effective and necessary measure for animal disease control.

[0003] Traditional methods for virus eradication include ultraviolet (UV) irradiation and disinfectant methods. Commonly used disinfectants include hydrogen peroxide, peracetic acid, chlorine dioxide, sodium hypochlorite, hypochlorous acid, sodium dichloroisocyanurate, alcohol (75%), glutaraldehyde, and quaternary ammonium salts. UV irradiation can oxidize the nucleic acid and protein amino acids of viruses, leading to viral death. The reactive oxygen species and reactive chlorine species in peroxides and chlorine-containing disinfectants can penetrate the cell wall into the cell to oxidize the phosphate dehydrogenase in viruses, causing an imbalance in sugar metabolism and leading to pathogen death. Alcohol (75%), glutaraldehyde, and quaternary ammonium salts can destroy the outer protein envelope of viruses, causing their genetic material to leak out and become inactive. UV irradiation requires clearing the disinfection site to prevent harm to humans and livestock. Although high concentrations of peroxides and chlorine-containing disinfectants have good sterilization effects, they pose a problem of damage to biomass caused by oxygen free radicals and hypochlorous acid.

[0004] Sodium chloride is widely used in the food, chlor-alkali, soda ash, and glass industries. It is mainly derived from sea salt and well salt. In my country, sodium chloride produced by the salt industry is primarily used in these sectors, resulting in relatively low added value. The application areas of sodium chloride need to be expanded to develop new applications and increase its added value. Currently, industrial sodium chloride particles range from several hundred micrometers to several millimeters in size. There are few technologies for preparing ultrafine sodium chloride powder. Furthermore, while ultrafine sodium chloride has been shown to be effective in treating asthma, its use for killing viruses has not yet been proposed. Existing patent application number ZL200910031335, entitled "A Method for Preparing Micron-Sized Ultrafine Sodium Chloride," lacks a corresponding antiviral mechanism (it cannot bind to and destroy the structure of viruses) and therefore cannot be used for virus killing. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to provide an avian influenza virus disinfectant that can effectively kill avian influenza virus, its use in killing the virus, and a method for killing the virus.

[0006] To solve the above-mentioned technical problems, the avian influenza virus disinfectant of the present invention uses sodium chloride aerosol with an average particle size of 800 nm and a particle size distribution between 500 nm and 8 μm directly as the avian influenza virus disinfectant.

[0007] Furthermore, a saturated sodium chloride aqueous solution and anhydrous ethanol are passed into the drying airflow of a spray dryer at a volume ratio of 1:1, and the sodium chloride aerosol is prepared by drying in the spray dryer.

[0008] Furthermore, the nitrogen flow rate in the spray dryer is 1.0 L / min.

[0009] Furthermore, the volume ratio of the saturated sodium chloride aqueous solution to the airflow in the spray dryer is 1:1000.

[0010] Furthermore, the drying gas flow of the spray dryer is nitrogen, and the drying temperature is 100-150°C. 0 C.

[0011] Furthermore, alcohols are added as antisolvents during the preparation of the sodium chloride aerosol.

[0012] Furthermore, the avian influenza virus in question is of the H9N2 subtype.

[0013] The use of an influenza virus disinfectant for virus killing: The influenza virus disinfectant is used for virus killing.

[0014] A method for killing influenza viruses using the above-mentioned influenza virus disinfectant, characterized in that: the dosage of the influenza virus disinfectant during the virus killing process is 15-30mg, and the killing time is 1-5min.

[0015] The advantages of this invention are: This invention combines antisolvent and spray drying methods with specific process parameters to prepare sodium chloride aerosol with an average particle size of 800 nm and a particle size distribution between 100 nm and 8 μm, which can be directly used as a disinfectant for avian influenza viruses. It has disinfection efficacy within a specific dosage and time range. High concentrations of sodium chloride solution can dehydrate bacterial cells through osmotic pressure difference, destroy their cell membrane structure, and lead to bacterial growth inhibition or death. In particular, it can achieve 100% inactivation of H9N2 subtype avian influenza virus with a small amount of sodium chloride aerosol and a short time, which is very effective. Attached Figure Description

[0016] Figure 1 This is a SEM image of the sodium chloride aerosol prepared according to the present invention. Detailed Implementation

[0017] The following detailed description, in conjunction with the accompanying drawings and specific embodiments, further illustrates the avian influenza virus disinfectant, its virus-killing applications, and virus-killing methods of the present invention.

[0018] The avian influenza virus disinfectant of this invention is prepared by passing a saturated sodium chloride aqueous solution and anhydrous ethanol in a 1:1 volume ratio into the drying gas stream of a spray dryer (with methanol or ethanol added as an anti-solvent). The resulting particles have an average size of 800 nm and a particle size distribution between 500 nm and 8 μm. Figure 1 Sodium chloride aerosol is used directly as a disinfectant against avian influenza virus. It works by altering the crystal structure or surface properties of sodium chloride, making it easier for the virus to bind and destroy its structure. The spray dryer has a drying chamber diameter and height of 0.5 x 0.5 m, a nitrogen flow rate of 1.0 L / min, a saturated sodium chloride aqueous solution to airflow volume ratio of 1:1000, and nitrogen as the drying gas. The drying temperature is 100-150 °C. 0 C, preferably 120 oC. Sodium chloride aerosol, used as a disinfectant against avian influenza viruses, is collected in a collector at the bottom of the spray dryer. The spray dryer is a device used to convert liquid materials into a dry, powdery product. Specific parameters of the spray dryer are crucial for producing sodium chloride aerosol as a disinfectant against avian influenza viruses. For example, the nitrogen flow rate is 1.0 L / min. This parameter determines the airflow velocity within the drying chamber, affecting the drying rate of the material. A faster flow rate helps improve drying efficiency, but an excessively fast flow rate may cause the material to be carried out of the drying chamber before it is fully dried. Another example is the saturated sodium chloride aqueous solution to airflow volume ratio of 1:1000, which controls the sodium chloride solution... The concentration of these parameters is crucial for forming aerosol particles of the desired size and uniformity. A suitable ratio ensures the generated aerosol particles have an ideal disinfection effect. Furthermore, using nitrogen as the drying gas stream and operating at 100-150°C (preferably 120°C) ensures sufficient drying of sodium chloride without causing thermal decomposition or destruction of active ingredients. Appropriate temperature contributes to improved product quality and stability. Verification has shown that the selection and optimization of these parameters are essential for producing sodium chloride aerosol products with highly effective disinfection capabilities, demonstrating the application value of spray drying technology in pharmaceutical and hygiene product manufacturing. Precise control of these parameters can improve product quality and production efficiency while ensuring product safety and efficacy. Especially for particle size control, precisely controlling the particle size of sodium chloride aerosol to an average of 800 nm with a distribution between 100 nm and 8 μm using anti-solvent and spray drying methods requires fine-tuning of process parameters. This includes the selection of antisolvents, optimization of parameters such as spray drying temperature, flow rate, and pressure. Finally, the size of the drying chamber can be controlled within a diameter and height of 0.5 meters, which affects the airflow distribution and material residence time within the drying chamber. Appropriate dimensions ensure that the material can be evenly exposed to the drying airflow, thereby promoting uniform drying.

[0019] The use of the influenza virus disinfectant of the present invention for virus killing involves using the above-mentioned influenza virus disinfectant for virus killing.

[0020] Test of sodium chloride aerosol's effectiveness in killing avian influenza virus (H9N2 subtype) 1 mL of virus solution was evenly sprayed into a cell culture dish, followed by evenly spraying 5 mg, 15 mg, 20 mg, 35 mg, and 50 mg of sodium chloride aerosol particles. After reacting for 1 min and 5 min, the mixture was collected for HA titer determination. Each time gradient was repeated three times. The initial hemagglutination titer of avian influenza virus was 2. 6 .

[0021] The efficacy of sodium chloride aerosol in killing avian influenza virus (H9N2 subtype) was determined using the avian influenza virus hemagglutinin glycoprotein (HA) level assay. The HA level assay was performed as follows: Add 0.025 mL of virus solution to well 1 and mix thoroughly by pipetting 3-5 times. Add 0.025 mL of virus solution from well 1 to well 2, mix well, and then add 0.025 mL to well 3. Dilute 2-fold to well 11, and discard 0.025 mL from well 11. Well 12 is the negative control. Add 0.025 mL of PBS to each well. Add 0.025 mL of 1% (v / v) chicken red blood cell suspension to each well. Gently tap the reaction plate to mix the reaction mixture and incubate at room temperature (approximately 20°C) for 40 min. The result is determined when the red blood cells in the control well show a distinct button-like appearance. During the determination, the reaction plate is tilted at 60° and the presence or absence of teardrop-like flow of red blood cells is observed. The highest dilution factor with no teardrop-like flow (100% agglutination) is determined as the blood coagulation titer.

[0022] The following examples demonstrate its effectiveness: Application Example 1 1 mL of avian influenza virus (H9N2 subtype) was evenly sprayed into a cell culture dish, followed by 5 mg of sodium chloride aerosol powder. After reacting for 1 min and 5 min, the mixture was collected for HA titer analysis, which showed a titer of 2. 6 .

[0023] Application Example 2 1 mL of avian influenza virus (H9N2 subtype) was evenly sprayed into a cell culture dish, followed by 15 mg of sodium chloride aerosol powder. After reacting for 1 min and 5 min, the mixture was collected for HA titer analysis, which showed a titer of 2. 6 With 2 5 .

[0024] Application Example 3 1 mL of avian influenza virus (H9N2 subtype) was evenly sprayed into a cell culture dish, followed by 20 mg of sodium chloride aerosol powder. After reacting for 1 min and 5 min, the mixture was collected and the HA titer was measured to be 2. 5.5 With 2 4 .

[0025] Application Example 4 1 mL of avian influenza virus (H9N2 subtype) was evenly sprayed into a cell culture dish, followed by 35 mg of sodium chloride aerosol powder. After reacting for 1 min and 5 min, the mixture was collected for HA titer analysis, which showed a titer of 2. 3 With 2 1 .

[0026] Application Example 5 1 mL of avian influenza virus (H9N2 subtype) was evenly sprayed into a cell culture dish, followed by 50 mg of sodium chloride aerosol powder. After reacting for 1 min and 5 min, the mixture was collected and the HA titer was 0.

[0027] Application Example 6 1 mL of avian influenza virus (H9N2 subtype) was evenly sprayed into a cell culture dish, followed immediately by 50 mg of sodium chloride powder with an average particle size of 550 μm. After reacting for 1 min and 5 min, the mixture was collected for HA titer analysis, which showed a titer of 2. 6 .

[0028] Table 1. Effects of sodium chloride aerosol on hemagglutination titer of avian influenza virus (H9N2 subtype) The experimental data above show that when the influenza virus disinfectant of this invention is used to kill avian influenza virus (H9N2 subtype), it exhibits a killing effect at a dosage of 15 mg and a killing time of 5 minutes. At a sodium chloride aerosol dosage of 30 mg and a killing time of 1 minute, the hemagglutination titer (HA) value increases from 2... 6 The value dropped to 0, demonstrating highly efficient avian influenza virus (H9N2 subtype) eradication characteristics.

[0029] Of course, the above description is not intended to limit the present invention, and the present invention is not limited to the examples given above. Any changes, modifications, additions or substitutions made by those skilled in the art within the scope of the present invention should also fall within the protection scope of the present invention.

Claims

1. A disinfectant for avian influenza viruses, characterized in that: Sodium chloride aerosol with an average particle size of 800 nm and a particle size distribution between 500 nm and 8 μm was directly used as a disinfectant against avian influenza virus.

2. The avian influenza virus disinfectant according to claim 1, characterized in that: A saturated sodium chloride aqueous solution and anhydrous ethanol were passed into the drying airflow of a spray dryer at a volume ratio of 1:1, and the sodium chloride aerosol was prepared by drying in the spray dryer.

3. The avian influenza virus disinfectant according to claim 2, characterized in that: The nitrogen flow rate in the spray dryer is 1.0 L / min.

4. The avian influenza virus disinfectant according to claim 2 or 3, characterized in that: The volume ratio of the saturated sodium chloride aqueous solution to the airflow in the spray dryer is 1:1000.

5. The avian influenza virus disinfectant according to claim 4, characterized in that: The spray dryer uses nitrogen as the drying gas flow and the drying temperature is 100-150°C. 0 C.

6. The avian influenza virus disinfectant according to claim 1, 2 or 4, characterized in that: Alcohols are added as antisolvents during the preparation of sodium chloride aerosols.

7. The avian influenza virus disinfectant according to claim 6, characterized in that: The avian influenza virus in question is of the H9N2 subtype.

8. The use of an influenza virus disinfectant for virus killing, wherein the influenza virus disinfectant according to any one of claims 1-7 is used for virus killing.

9. A method for killing viruses using an influenza virus disinfectant as described in any one of claims 1-7, characterized in that: The influenza virus disinfectant is used at a dosage of 15-30 mg during the virus killing process, and the killing time is 1-5 minutes.

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