Composite bio-enzyme air disinfection method based on large-flow concentration and enrichment combined with aerosol-hydrosol transfer

Abstract

According to the composite bio-enzyme air disinfection method based on large-flow concentration and enrichment combined with aerosol-hydrosol transfer provided by the invention, through large-flow enrichment combined with aerosol-hydrosol transfer, bio-aerosol particles in a large-volume space can be transferred into a small-volume liquid phase within a short time; enriched microorganisms are intensively and efficiently killed through the composite biological enzyme, so that efficient indoor air disinfection is realized. The biological enzyme component can be atomized into the air through re-aerosolization in the sampling and transferring process from the gas phase to the liquid phase, so that a certain degree of direct air disinfection effect can be realized. Microbial particles escaping after aerosolization again are also killed or inhibited due to the combination of the composite biological enzyme component. Due to the fact that the composite biological enzyme safe to human bodies is used as the main component of the disinfectant, the method basically has no influence on the air environment while efficient disinfection is achieved, and pathogens in indoor air can be efficiently and safely disinfected under the condition that people flow.

Description

Technical Field

[0001] The present invention belongs to the technical field of air disinfection, and in particular relates to a composite bio-enzyme air disinfection method based on large-flow concentration and enrichment combined with aerosol-hydrosol transfer. Background Art

[0002] Indoor air in homes, offices, hospitals, high-speed trains, airplanes, ships, and submarines primarily contains microbial aerosols, including viruses, bacteria, and mold. The types, quantities, and proportions of these bioaerosols vary across different indoor environments. Some microorganisms are pathogens. Even at very low concentrations, they can enter the body through breathing or contact, causing infection and posing a significant threat to human health. Furthermore, their accumulation in poorly ventilated indoor air can contribute to the spread of pathogens. Therefore, air disinfection in relatively confined spaces is essential.

[0003] Methods and technologies for eliminating airborne microorganisms can generally be categorized into three main categories: physical, chemical, and biochemical. These include four physical disinfection technologies: thermal disinfection, high-efficiency filtration, ultraviolet disinfection, and electrostatic adsorption; two chemical disinfection technologies: ozone disinfection and disinfectant atomization; three physical-chemical disinfection technologies: high-voltage discharge, photocatalysis, and nanosilver technology; and one biochemical disinfection technology: lysozyme. These technologies vary in their disinfection capabilities, energy consumption, treatment targets, and treatment scopes.

[0004] Thermal disinfection technology is widely used and reliable, but it is limited by the heat resistance of the objects and the location of its application. High-efficiency filtration disinfection uses physical retention to remove airborne pathogens. Its advantages are that it can be removed at room temperature and is simple to use. However, its disadvantages are that it only filters and does not kill microorganisms, potentially causing secondary contamination. Ultraviolet disinfection is the most commonly used bioaerosol disinfection technology for confined spaces. It is fast and efficient, effectively inactivating nearly all bacteria and viruses. Although it is relatively well-established, ultraviolet light is harmful to the human body and should not be used directly in areas where people are active. Electrostatic adsorption technology offers advantages such as high adsorption efficiency, low pressure drop, and low cost, but the corona discharge process may generate large amounts of ozone. Ozone disinfection is thorough, broad-spectrum, and highly effective, with significant killing effects on all bacteria and viruses. However, high residual ozone concentrations can be harmful to the human body. Disinfectant atomization is effective, but it is prone to reagent leakage during the disinfection process, inevitably resulting in chemical residues that can have side effects on the human body. It is generally not suitable for disinfecting precision instruments, tableware, etc. High-voltage discharge technology can generate various active substances, such as plasma and negative ions, under different conditions, all of which can be used to disinfect pathogenic microorganisms. However, this process also produces harmful byproducts such as ozone and nitrogen oxides. Photocatalytic disinfection primarily relies on the reaction of charge carriers generated by photocatalytic materials absorbing light radiation with O₂, H₂O, and other substances adsorbed on the material's surface, generating reactive free radicals that oxidatively degrade microorganisms. However, current photocatalytic materials have a narrow light response range, high charge carrier recombination rates, and most require ultraviolet light to function. Furthermore, their catalytic performance gradually declines due to factors such as inherent defects and the surface environment. Silver nanoparticle technology offers advantages such as safety, efficiency, environmental friendliness, and broad spectrum, but Ag nanoparticles still agglomerate and leak within the matrix, reducing the material's recyclability and effectiveness. Lysozyme is a natural antimicrobial agent with very high antimicrobial efficacy and biosafety. However, since pure lysozyme requires a liquid or near-liquid environment for application and suffers from drawbacks such as poor stability, reusability, and leaching, it is currently difficult to achieve large-scale air disinfection. However, due to its good safety, it can be predicted that lysozyme technology will have unique advantages and broad application prospects in the field of air disinfection.

[0005] In summary, high-efficiency filtration and electrostatic adsorption are economical and affordable, but they can only achieve the interception and sedimentation of pathogenic microorganisms and cannot disinfect and inactivate them. Ultraviolet disinfection, ozone disinfection, and disinfectant atomization are simple and convenient, but the byproducts produced by them or during use can be harmful to the human body, so they can only be used when no one is around. High-voltage discharge, photocatalysis, nanosilver, and lysozyme technologies are efficient and safe, but their range of action is limited and cannot achieve large-scale disinfection. In summary, it can be seen that a single disinfection technology is often difficult to achieve rapid and efficient disinfection of pathogenic microorganisms. Although multiple disinfection technologies have been combined for air disinfection, they have shortcomings and drawbacks such as limited efficiency, unstable catalyst activity, and unclear biological toxicity. Therefore, the development of safe and efficient multi-factor, multi-functional, and high-efficiency methods and technologies for indoor air disinfection is an important future development direction and has great significance for epidemic prevention and control, economic and social stability, and the construction of a national biosafety system.

[0006] Based on the urgent need to develop safe and efficient indoor air disinfection methods, exploring and establishing green and environmentally friendly sterilization and disinfection methods that do not harm human health, so as to achieve high-efficiency indoor air sterilization and disinfection with little or no impact on the air environment, thereby disinfecting pathogens in indoor air when there is personnel flow, is an urgent problem to be solved. Summary of the Invention

[0007] (1) Technical issues to be resolved

[0008] The present invention proposes a composite bio-enzyme air disinfection method based on large-flow concentration and enrichment combined with aerosol-hydrosol transfer. Through large-flow concentration and enrichment combined with aerosol-hydrosol transfer technology, the bioaerosol particles in a large volume space are transferred to the liquid phase in a short time, and the enriched microorganisms are concentratedly and efficiently removed by composite bio-enzymes, so as to solve the technical problem that it is difficult to strike a balance between high efficiency and safety in current indoor air disinfection methods, achieve high-efficiency air disinfection while having little or basically no impact on the air environment, thereby disinfecting pathogens in indoor air under the condition of personnel flow.

[0009] (2) Technical solution

[0010] In order to solve the above technical problems, the present invention proposes a composite bio-enzyme air disinfection method based on large-flow concentration and enrichment combined with aerosol-hydrosol transfer. The composite bio-enzyme air disinfection method is:

[0011] S1. Concentrate and enrich aerosol particles in the air at a high flow rate;

[0012] S2. Perform aerosol-hydrosol transfer on the concentrated and enriched aerosol particles and sample them into a sampling sterilization solution;

[0013] S3. Use the liquid phase composite bio-enzyme in the sampled sterilization liquid to carry out sterilization and disinfection.

[0014] Furthermore, step S1 specifically includes: using a large flow concentrator to concentrate and enrich aerosol particles in the air.

[0015] Furthermore, through step S1, the concentration and enrichment of aerosol particles with a particle size of 0.5 μm or more is achieved.

[0016] Furthermore, through step S1, aerosol particles with a particle size of 0.5 μm or more are concentrated and enriched by 4 to 15 times.

[0017] Furthermore, the flow rate of the large-flow concentrator is 3600 L / min, and the flow rate of the effective small-flow secondary airflow after concentration and enrichment is controlled at 250 L / min to 300 L / min.

[0018] Furthermore, step S2 specifically includes: sampling the small-flow secondary airflow for concentrating and enriching aerosol particles into the sampling sterilization liquid through an aerosol sampler.

[0019] Furthermore, the aerosol sampler is a wetted wall cyclone aerosol sampler.

[0020] Furthermore, the sampling flow rate of the wetted wall cyclone aerosol sampler is 100 L / min to 300 L / min.

[0021] Furthermore, the sampling sterilization liquid contains lysozyme, lysostaphin, antimicrobial peptides and chlorhexidine acetate; the collected microorganisms are automatically killed through the synergistic enzymatic action of lysozyme and lysostaphin, the antimicrobial action of antimicrobial peptides and the antimicrobial action of chlorhexidine acetate.

[0022] Furthermore, each liter of the sampled sterilization solution contains 2 g of lysozyme, 0.005 g of lysostaphin, 0.5 g of type A nisin, 1 g of ε-polylysine, 1 g of chlorhexidine acetate, and 1.5 g of auxiliary material trehalose, and the solvent is water.

[0023] (3) Beneficial effects

[0024] The present invention proposes a composite bio-enzyme air disinfection method based on large-flow concentration and enrichment combined with aerosol-hydrosol transfer. Through large-flow enrichment combined with aerosol-hydrosol transfer, the bioaerosol particles in a large volume space can be transferred to a small volume liquid phase in a short time, and the enriched microorganisms can be concentrated and efficiently killed by the composite bio-enzyme, thereby achieving high-efficiency indoor air disinfection. The re-aerosolization during the gas-to-liquid phase sampling transfer process can cause the bio-enzyme components to be atomized into the air, thereby also achieving a certain degree of direct air disinfection effect. The microbial particles that escape due to re-aerosolization are also disinfected or inhibited due to binding to the composite bio-enzyme components. Since a composite bio-enzyme that is safe for the human body is used as the main component of the disinfectant, this method has basically no impact on the air environment while achieving efficient disinfection, so that pathogens in indoor air can be efficiently and safely disinfected in the presence of personnel flow, and has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 A schematic diagram of the principle of the method proposed in the present invention;

[0026] In the figure: A is the aerosol flow before high-flow enrichment; B is the concentrator; C is the low-flow concentrated aerosol flow after concentration and enrichment; D is the wetted wall cyclone sampler that converts aerosol into hydrosol (the sampler contains a sampling sterilization solution with a complex biological enzyme); E is the aerosol flow generated by the re-aerosolization during the sampling process;

[0027] Figure 2 The test results for verifying the sterilization and disinfection effect of artificial bioaerosol enrichment and concentration in the embodiment of the present invention are as follows;

[0028] In the figure: A is the concentration change of Escherichia coli aerosol sampling and killing before and after, where 1 is the concentration before sampling and killing, and 2 is the concentration after sampling and killing; B is the concentration change of Staphylococcus aureus aerosol sampling and killing before and after, where 1 is the concentration before sampling and killing, and 2 is the concentration after sampling and killing; C is the concentration change of Candida albicans aerosol sampling and killing before and after, where 1 is the concentration before sampling and killing, and 2 is the concentration after sampling and killing; the vertical axis is the concentration of biological aerosol, the unit is CFU / m 3 . DETAILED DESCRIPTION

[0029] In order to make the purpose, content and advantages of the present invention more clear, the specific implementation methods of the present invention are further described in detail below with reference to the accompanying drawings and examples.

[0030] This embodiment proposes a composite bio-enzyme air disinfection method based on large-flow concentration and enrichment combined with aerosol-hydrosol transfer, which specifically includes the following steps:

[0031] S1. High-flow concentration and enrichment of aerosol particles in the air

[0032] A large-flow concentrator with a flow rate of 3600L / min is used to concentrate and enrich the aerosol particles in the air. The flow rate of the effective small-flow secondary airflow after concentration and enrichment is controlled at 250L / min~300L / min, achieving 4~15 times concentration and enrichment of aerosol particles with a particle size of 0.5μm or more.

[0033] S2. Transfer the concentrated aerosol particles to a hydrosol and sample them into the sampling sterilization solution.

[0034] The small-flow secondary airflow that concentrates and enriches aerosol particles is sampled into the sampling sterilization liquid through a wetted wall cyclone aerosol sampler (sampling flow rate is 100L / min to 300L / min).

[0035] S3. Use liquid phase compound enzymes in the sampled sterilization solution for sterilization and disinfection

[0036] The sampling sterilization solution contains components of complex biological enzymes such as lysozyme, lysostaphin, and various antimicrobial peptides, as well as the broad-spectrum antimicrobial agent chlorhexidine acetate, as well as protective excipients such as water and trehalose. While sampling, the collected microorganisms are automatically killed through the synergistic enzymatic hydrolysis of lysozyme and lysostaphin, the antimicrobial action of the antimicrobial peptides, and the antimicrobial action of chlorhexidine acetate. In this embodiment, the sampling sterilization solution is formulated as follows: per liter of sampling sterilization solution, it contains 2g of lysozyme, 0.005g of lysostaphin, 0.5g of type A nisin, 1g of epsilon-polylysine, 1g of chlorhexidine acetate, and 1.5g of trehalose (excipient), with water as the solvent.

[0037] Verification of the sterilization effect of compound bio-enzyme sampling sterilization solution:

[0038] The bactericidal effect of the sampled bactericidal solution was verified according to the following steps. (1) Preparation of bacterial suspension: Staphylococcus aureus, Escherichia coli and Candida albicans were isolated and cultured separately, and fresh slant cultures of each strain for 24 hours were taken, washed with 0.03 mol / L phosphate buffer (PBS, pH 7.2-7.4) and diluted to a concentration of 10 5 ~10 6cfu / mL bacterial suspension. (2) Sterilization test: Pipette 0.5mL of the test bacterial solution into 4.5mL of the sampled sterilization solution, mix well, and allow to act for 1min, 5min, and 10min respectively. Immediately after that, take 0.5mL of the above mixture, add 4.5mL of neutralizer (phosphate buffer containing 0.5% sodium thiosulfate + 3% Tween-80 + 0.5% lecithin, 0.03M, pH 7.2-7.4), mix well, and allow to act for 10min. Pipette 1mL of the test bacterial solution and the mixture after the action for plate culture to determine the number of colonies and calculate the sterilization efficiency. Repeat the test 3 times. The results are shown in the table below. It can be seen that the composite bio-enzyme sampling sterilization solution of this embodiment acts on Escherichia coli, Staphylococcus aureus and Candida albicans in the suspension for 1 minute, 5 minutes and 10 minutes, respectively. The average sterilization rates of Escherichia coli are 96.64%, 98.61% and 100%, respectively, while the average sterilization rates of Staphylococcus aureus and Candida albicans are both 100%.

[0039]

[0040] Lysozyme catalyzes the hydrolysis of the β-1,4 glycosidic bond between N-acetylmuramic acid and N-acetylglucosamine in the peptidoglycan of the bacterial cell wall, thereby causing the cell wall to rupture, the contents to overflow and the bacteria to lyse; lysostaphin catalyzes the cleavage of the pentapeptide Gly-Gly bond in the peptidoglycan of the cell wall; antimicrobial peptides are a class of small peptide substances with microbial killing activity, such as nisin, bisin, bacitracin, iseganan, indolicidin, omiganan, neuprex and ε-polylysine, each with its own unique mechanism of action. For example, the antibacterial mechanism of nisin is to form potential-dependent ion channels on the plasma membrane of sensitive bacteria, allowing ions and small molecules to quickly overflow, leading to bacterial death; chlorhexidine acetate achieves a broad-spectrum bacterial killing effect by changing the permeability of the bacterial cell membrane. The lysozyme, lysostaphin, nisin, ε-polylysine and chlorhexidine acetate composite enzyme combination adopted in the present embodiment are compounded, by the enzymolysis of bacterial cell wall and the synergistic effect that changes cell membrane permeability, broaden the bactericidal spectrum, improve antibacterial ability, realize the broad spectrum strong killing action to Gram-positive bacteria, Gram-negative bacteria and fungi.Each component of composite enzyme has no toxic effect on human cells and tissue, and has been widely used in flushing, disinfection and treatment of medical field.Even therefore also harmless to human body even if a small amount of being aerosolized and released into the air in the sampling sterilization process, and each component of composite enzyme can also bring into play certain bactericidal action in air.

[0041] Verification test of artificial bioaerosol enrichment and concentration sterilization and disinfection effect:

[0042] Using 8m 3The bactericidal and disinfection effect of this method was tested by generating bacterial aerosols in the bioaerosol test cabin. A high-flow concentrator and a wet-wall cyclone sampler (complex biological enzymes were added to the sampling liquid) were connected and placed in the bioaerosol generation test cabin. After the cabin completed self-cleaning, Escherichia coli, Staphylococcus aureus, and Candida albicans aerosols were generated in the cabin respectively. The wet-wall cyclone sampler was used to collect samples to determine the concentration of active bacterial aerosols. After the sampling was completed, the high-flow concentrator and the wet-wall cyclone sampler were turned on for continuous sampling and sterilization for 10 minutes. After the sampling and sterilization was completed, the wet-wall cyclone sampler was used to sample the nutrient agar plate to determine the concentration of active bacterial aerosols.

[0043] The results are as follows Figure 2 As shown in the figure, the aerosol concentrations of active Escherichia coli, Staphylococcus aureus, and Candida albicans before sampling and sterilization were 740667 CFU / m 3 、1386667CFU / m 3 and 416506 CFU / m 3 After 10 minutes of continuous sampling and sterilization, the concentration of active bacteria aerosol decreased to 33600 CFU / m 3 、6380CFU / m 3 and 9389 CFU / m 3 It can be seen that the concentration of live bacteria in the sample culture was greatly reduced after continuous sampling and sterilization for 10 minutes. 3 The removal rates of Escherichia coli, Staphylococcus aureus and Candida albicans in the cabin reached 95.46%, 95.40% and 97.75% respectively within 10 minutes. This method has obvious effect on the enrichment and concentration of artificial bioaerosols for sterilization and disinfection.

[0044] Verification test of sterilization and disinfection effect of air enrichment and concentration in indoor confined spaces:

[0045] Using 8m 3 The method was tested for its effectiveness in sterilizing and disinfecting indoor air in a bioaerosol test chamber. After the chamber was self-cleaned, the test chamber door was opened to connect the chamber to the laboratory. After equilibrium was reached, the door was closed and sampling was performed using a six-stage Andersen sampler in conjunction with a nutrient agar plate for 10 minutes. The total number of colonies was cultured and counted to obtain the concentration of culturable microorganisms. After sampling, the high-flow concentrator and wet-wall cyclone sampler were turned on for continuous sampling and sterilization for 10 minutes. After sampling and sterilization, sampling was performed using a six-stage Andersen sampler in conjunction with a nutrient agar plate for 10 minutes. The total number of colonies was cultured and counted to obtain the concentration of culturable microorganisms. This was used to calculate the removal rate of culturable microorganisms in the chamber. The test was conducted five times, once per day.

[0046] The results are shown in the table below. It can be seen that after continuous 10 minutes of sampling and sterilization, the concentration of culturable microorganisms in the air of the closed cabin was greatly reduced. 3The results of five tests on the removal rate of culturable microorganisms in the cabin air within 10 minutes were 90.28%, 88.65%, 90.57%, 87.3% and 90%, with an average of 89.36%. This method has obvious effects on the enrichment, concentration, sterilization and disinfection of actual indoor air microorganisms.

[0047]

[0048]

[0049] According to the embodiments, the present invention proposes a composite bio-enzyme indoor air sterilization and disinfection method based on high-flow concentration and enrichment combined with aerosol-hydrosol transfer. Through high-flow enrichment combined with high-efficiency aerosol-hydrosol transfer, bioaerosol particles in a large volume can be transferred to a small liquid phase in a short period of time. The composite bio-enzyme concentrates and efficiently removes artificial microbial aerosols such as Staphylococcus aureus, Escherichia coli, and Candida albicans that have been enriched in the liquid phase. It also achieves a high removal rate for actual airborne microorganisms in confined spaces, achieving highly efficient indoor air sterilization and disinfection. Re-aerosolization during the gas-to-liquid phase sampling and transfer process allows a small amount of bio-enzyme components to be atomized into the air, thereby also achieving a certain degree of direct air disinfection. Microbial particles that escape after re-aerosolization are also disinfected or inhibited due to binding to the composite bio-enzyme components. Because the composite bio-enzyme, which is safe for human health, is used as the main component of the disinfectant, this method achieves efficient disinfection while having little impact on the air environment. This allows for efficient and safe disinfection of pathogens in indoor air even in the presence of human traffic, and has broad application prospects.

[0050] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the technical principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A composite bio-enzyme air disinfection method based on large-flow concentration and enrichment combined with aerosol-hydrosol transfer, characterized in that: The composite biological enzyme air disinfection method is: S1. Concentrate and enrich aerosol particles in the air at a large flow rate; S2. Perform aerosol-hydrosol transfer on the concentrated and enriched aerosol particles and sample them into a sampling sterilization solution; S3. Use the liquid phase composite bio-enzyme in the sampled sterilizing liquid to carry out sterilization and disinfection.

2. The composite bio-enzyme air disinfection method based on large flow concentration and enrichment combined with aerosol-hydrosol transfer as claimed in claim 1, characterized in that: Step S1 specifically includes: using a large flow rate concentrator to concentrate and enrich aerosol particles in the air.

3. The composite bio-enzyme air disinfection method based on large flow concentration and enrichment combined with aerosol-hydrosol transfer as claimed in claim 2, characterized in that: Through step S1, the aerosol particles with a particle size of 0.5 μm or more are concentrated and enriched.

4. The composite bio-enzyme air disinfection method based on large flow concentration and enrichment combined with aerosol-hydrosol transfer as claimed in claim 3 is characterized in that: Through step S1, 4 to 15 times concentration and enrichment of aerosol particles with a particle size of 0.5 μm or more is achieved.

5. The composite bio-enzyme air disinfection method based on large flow concentration and enrichment combined with aerosol-hydrosol transfer as claimed in claim 2, characterized in that: The flow rate of the large-flow concentrator is 3600 L / min, and the flow rate of the effective small-flow secondary airflow after concentration and enrichment is controlled at 250 L / min to 300 L / min.

6. The composite bio-enzyme air disinfection method based on large flow concentration and enrichment combined with aerosol-hydrosol transfer as claimed in claim 1, characterized in that: Step S2 specifically includes: sampling a small-flow secondary airflow for concentrating and enriching aerosol particles into a sampling sterilization liquid through an aerosol sampler.

7. The composite bio-enzyme air disinfection method based on large flow concentration and enrichment combined with aerosol-hydrosol transfer as claimed in claim 6, characterized in that: The aerosol sampler is a wet wall cyclone aerosol sampler.

8. The composite bio-enzyme air disinfection method based on large flow concentration and enrichment combined with aerosol-hydrosol transfer as claimed in claim 7, characterized in that: The sampling flow rate of the wet wall cyclone aerosol sampler is 100L / min to 300L / min.

9. The composite bio-enzyme air disinfection method based on large flow concentration and enrichment combined with aerosol-hydrosol transfer as claimed in claim 1, characterized in that: The sampling sterilization liquid contains lysozyme, lysostaphin, antimicrobial peptide and chlorhexidine acetate; the collected microorganisms are automatically killed through the synergistic effects of the enzymatic hydrolysis of lysozyme and lysostaphin, the antimicrobial effect of the antimicrobial peptide and the antimicrobial effect of chlorhexidine acetate.

10. The composite bio-enzyme air disinfection method based on large flow concentration and enrichment combined with aerosol-hydrosol transfer as claimed in claim 9, characterized in that: Each liter of sampling sterilization solution contains 2g of lysozyme, 0.005g of lysostaphin, 0.5g of type A nisin, 1g of ε-polylysine, 1g of chlorhexidine acetate, and 1.5g of auxiliary material trehalose, and the solvent is water.

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