Nanosalt particle disinfectant for inhibiting pathogenic fungi and use of the same

Abstract

This invention discloses a nano-salt particle disinfectant for inhibiting pathogenic fungi. It comprises nano-salt particles with a particle size distribution between 400 and 1000 nm. The disinfectant is prepared by atomizing a 0.1 M sodium chloride aqueous solution into aerosol droplets and introducing them into a reactor, with the reaction temperature controlled between 20°C and 30°C. The advantages are: by employing an antisolvent precipitation method combined with high-energy nano-sizing technology, sodium chloride powder with uniform particle size and nanoscale was successfully prepared; the antifungal efficacy of this novel nano-salt particle as the core component of the atomized disinfectant was systematically evaluated, targeting major human pathogenic fungi.

Description

Technical Field

[0001] This invention belongs to the field of nano-salt particle disinfectants and their applications, and in particular relates to a nano-salt particle disinfectant for inhibiting pathogenic fungi and its application to pathogenic fungi. Background Technology

[0002] The global incidence of invasive fungal infections has been steadily rising in recent decades, a worrying trend primarily attributed to the expanding immunocompromised population, including those undergoing chemotherapy, organ transplants, and those infected with HIV or living with AIDS. These epidemiological changes pose a significant challenge to global public health systems, further compounded by the limited availability of effective systemic antifungal agents. An article published in the 2023 issue of the Chinese Journal of Biomedicine, "Diagnosis of Invasive Fungal Infections: Challenges and Recent Advances," reveals that the clinical treatment of invasive fungal infections faces multiple obstacles, including difficulty in accessing antifungal drugs, significant drug-related toxicities, and the increasing prevalence of antifungal resistance in major pathogenic fungi (such as Aspergillus, Candida, and Cryptococcus).

[0003] The 2024 IJMS journal article, "Human-Fungal Pathogen Interactions from the Perspective of Immunoplasmic Analysis," reveals that the respiratory tract is the primary route of invasion for most invasive fungal pathogens. Inhalation of airborne conidia or yeast cells can trigger primary pulmonary infections and may further disseminate, leading to life-threatening systemic infections. Notably, data from the 2021 North American Infectious Disease Clinic, by Cadena J, Thompson GR, and Patterson TF, indicates that invasive fungal infections caused by Aspergillus fumigatus have a mortality rate exceeding 50% in immunocompromised patients, and in some endemic areas, resistance to azole antifungals has exceeded 20%. Therefore, implementing effective air purification measures in clinical settings and high-risk environments has become a crucial aspect of controlling such infections. However, the development of safe and efficient aerosol disinfectants lags behind actual clinical needs. Most traditional disinfectants are unsuitable for aerosol applications due to potential respiratory toxicity, irritation, or poor stability in aerosol form, thus representing a significant weakness in infection prevention strategies.

[0004] Nanomaterials, with their size-dependent physical interactions, offer potentially revolutionary solutions for antibacterial therapy. While metallic nanoparticles such as silver and zinc oxide possess certain antifungal activity, their potential biocompatibility issues and complex preparation processes limit their clinical translation. Sodium chloride, as an important inorganic salt, boasts significant advantages such as high biocompatibility, wide availability, and low cost. It plays a crucial role in various physiological processes, particularly in maintaining fluid homeostasis. In addition to its physiological functions, hypertonic sodium chloride solutions have been widely demonstrated to possess potent antibacterial and disinfectant effects, primarily through osmotic pressure inducing microbial cell dehydration and lysis. Fully utilizing its inherent disinfectant properties and developing it into a safe, controllable, and nebulizable formulation holds promise for significantly revolutionizing environmental fungal control strategies. However, in the fields of materials science and disinfection technology, successfully preparing water-soluble salts such as sodium chloride into stable nanoparticles suitable for aerosol delivery remains a key and unresolved technical challenge. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to provide a nano-salt particle disinfectant that can effectively inhibit pathogenic fungi and its application to pathogenic fungi.

[0006] To solve the above-mentioned technical problems, the present invention provides a nano-salt particle disinfectant for inhibiting pathogenic fungi, comprising nano-salt particles with a particle size distribution between 400 and 1000 nm. The nano-salt particle disinfectant for inhibiting pathogenic fungi is obtained by atomizing a 0.1 M sodium chloride aqueous solution into aerosol droplets and introducing them into a reactor, while controlling the reaction temperature between 20°C and 30°C.

[0007] The pathogenic fungi include Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger, Candida albicans, and Cryptococcus neoformans.

[0008] The purpose of the nano-salt particle disinfectant for inhibiting pathogenic fungi is to inhibit pathogenic fungi.

[0009] The application of nano-salt particle disinfectant for inhibiting pathogenic fungi involves uniformly spraying the nano-salt particle disinfectant into a disinfection space to achieve a concentration of 20-400 nano-salt particles / cm³ with a particle size of 400-1000nm.

[0010] The nano-salt particles are uniformly sprayed using an electric powder sprayer.

[0011] Advantages of this invention:

[0012] By employing an antisolvent precipitation method combined with high-energy nanotechnology, sodium chloride powder with uniform particle size and nanoscale structure was successfully prepared. The antifungal efficacy of this novel nano-salt particle as a core component of an atomized disinfectant was systematically evaluated, targeting major human pathogenic fungi, including *Aspergillus spp.*, *Candida albicans*, and *Cryptococcus neoformans*. Furthermore, its bactericidal mechanism against highly resistant *Aspergillus fumigatus* conidia was further investigated. The experimental results provide strong evidence that the nano-salt particles physically disrupt the structural integrity of fungal cell walls and cell membranes, inducing pore formation, leading to the leakage of key intracellular substances, and ultimately causing cell lysis and death. This study not only systematically evaluates the disinfection potential of a groundbreaking nano-formulation but also explores its practical application value in controlling airborne fungal contamination. Attached Figure Description

[0013] Figure 1 This invention relates to the dynamic monitoring of particle size during the preparation of nano-salt particles in a nano-salt particle disinfectant for inhibiting pathogenic fungi.

[0014] Figure 2 The present invention provides a nano-salt particle disinfectant for inhibiting pathogenic fungi, demonstrating that the nano-salt is active against both azole-sensitive and resistant Aspergillus fumigatus spores. A and D are photographs of clinically and environmentally derived Aspergillus fumigatus fungal colonies; B and E are statistical data on counts after dilution and application; C and F are evaluations of the nano-salt's anti-Aspergillus fumigatus activity based on survival rate.

[0015] Figure 3 The present invention provides a nano-salt particle disinfectant for inhibiting pathogenic fungi, which shows that the nano-salt has antifungal activity and exhibits a strong inhibitory effect on Aspergillus flavus and Aspergillus niger. In the figure, A is a photograph of Aspergillus flavus and Aspergillus niger colony-forming units, B is the data after normalization based on the CFU value after dilution and coating, and C is the survival rate of Aspergillus flavus and Aspergillus niger.

[0016] Figure 4 The present invention provides a nano-salt particle disinfectant for inhibiting pathogenic fungi, which shows that the nano-salt has a bactericidal effect against key pathogens Candida albicans and Cryptococcus neoformans. Among them, A, B, and C demonstrate the antifungal effect of nano-salt against Candida albicans through CFU photograph evidence, CFU count statistics, and survival rate analysis; D, E, and F use the same procedure to evaluate the antifungal effect of nano-salt against Cryptococcus neoformans.

[0017] Figure 5 The present invention provides a nano-salt particle disinfectant for inhibiting pathogenic fungi, showing that nano-salt treatment leads to the breakage and structural destruction of Aspergillus fumigatus spores, scale bar = 10 μm;

[0018] Figure 6 CFW staining of the nano-salt particle disinfectant used in this invention for inhibiting pathogenic fungi showed that nano-salt treatment disrupted the cell wall integrity of Aspergillus fumigatus spores; in section A, CFW staining was used to evaluate the changes in the cell wall of Aspergillus fumigatus conidia and the fluorescence intensity of spores after treatment with different nano-salt concentrations; in section B, the fluorescence intensity of the conidia stained on the left side was calculated using ImageJ. Scale bar = 10 μm and 5 μm.

[0019] Figure 7 In this invention, the loss of cell membrane integrity of Aspergillus fumigatus spores treated with nano-salt was observed by PI staining in the nano-salt particle disinfectant used to inhibit pathogenic fungi. In this invention, A represents the activity of Aspergillus fumigatus conidia directly measured by PI staining; B represents the percentage of corresponding PI-positive cells. Scale bar = 10 μm. Detailed Implementation

[0020] The following detailed description, in conjunction with the accompanying drawings and specific embodiments, further illustrates the present invention's nano-salt particle disinfectant for inhibiting pathogenic fungi and its application to pathogenic fungi.

[0021] Example:

[0022] Atomized nano-salt particles were prepared by the following steps: A 0.1M sodium chloride aqueous solution was atomized to form aerosol droplets and introduced into a high-temperature reactor. By controlling the aerosol flow rate and reaction temperature, nano-salt particles with a particle size distribution between 400 and 1000 nm were obtained. These nano-salt particles can be used as a disinfectant to inhibit pathogenic fungi. The aerosol droplets undergo Brownian motion in the air, and the reaction temperature is controlled between 20℃ and 30℃, with an optimal temperature of 25℃. The nano-salt particles were then uniformly sprayed into a disinfection space, resulting in a number concentration of 20-400 nano-salt particles (400-1000 nm) per cm³. The spraying time was 5 seconds, and the flow rate was 4-80 g / (cm³·s).

[0023] In actual operation, with a spray time of 5 seconds and a flow rate of 1480~29600 particles / s, the measured 25,000 particles in the 400-1000 nm range corresponded to a peak number concentration of approximately 67.6 particles / cm³ in a 370 cm³ chamber.

[0024] The bactericidal effect was tested, as follows:

[0025] Culture of fungal strains: The fungal strains included *Aspergillus fumigatus*, *Aspergillus flavus*, *Aspergillus niger*, *Candida albicans*, and *Cryptococcus neoformans*. *Aspergillus fumigatus*, *Aspergillus flavus*, and *Aspergillus niger* were inoculated into YAG medium containing 2% glucose, 0.5% yeast extract, and trace elements to obtain fungal conidia. The fungal conidia included *Aspergillus fumigatus* conidia, *Aspergillus flavus* conidia, and *Aspergillus niger* conidia. *Candida albicans* and *Cryptococcus neoformans* were cultured in SDA medium containing 4% glucose, 1% peptone, and at a pH of 5.4-5.8 to obtain yeast cells. The culture temperature for both YAG and SDA media was 37°C.

[0026] Evaluation of antifungal activity: Nano-salt particles were uniformly sprayed for 5 seconds using an electric powder sprayer. The suspension of cultured fungal conidia or yeast cells was atomized and uniformly dispersed in a closed device using a medical nebulizer and mixed with nano-salt particles for 2 hours. The fungal cells after the reaction were collected and spread on a solid culture medium, incubated at 37°C, and counted. The colony forming units (cfu) were counted and the survival rate was calculated accordingly.

[0027] Flow cytometry: Aspergillus fumigatus conidia reacted with nano-salt particles at different mixing time points were collected, centrifuged at 10,000-13,000 rpm for 3-5 min, the supernatant was discarded, and DCFH-DA probe with a final concentration of 10 μM was added. The cells were incubated at 37ºC for 30-50 min, with the cells inverted every 5-10 min to ensure sufficient contact between the probe and the cells. The fungal cells were then washed 2-3 times with phosphate buffer to remove any probe that had not entered the cells. Finally, the cells were resuspended in 1 mL of phosphate buffer and analyzed using a flow cytometer with an excitation wavelength of 488 nm and an emission wavelength of 525 nm. The fluorescence intensity of at least 15,000 conidia was detected.

[0028] Scanning electron microscopy (SEM): Fresh, untreated Aspergillus fumigatus conidia and nano-salt treated Aspergillus fumigatus conidia were collected separately. After centrifugation at 10,000-13,000 rpm for 3-5 min, the supernatant was discarded, and the conidia were resuspended and fixed in 1 mL of 3% glutaraldehyde solution. The morphological changes of conidia under different treatments were observed using a Hitachi Regulus-8100 scanning electron microscope (Hitachi, Tokyo, Japan).

[0029] Calcium fluorescent white (CFW) staining: Aspergillus fumigatus conidia collected after reaction with nano-salt particles were transferred to a glass slide, and 150 μL of CFW staining solution with a concentration of 10 μg / mL was added to the surface. The slide was incubated at 37 ℃ for 5 min, the CFW staining solution was aspirated, and the fungal cells were washed 2-3 times with 1 mL of phosphate buffer. The samples were fixed with 4% paraformaldehyde for 30 min, a coverslip was placed on the glass slide, and filter paper was gently pressed to absorb excess moisture. The slide was then sealed with nail polish to obtain the treated conidia. The conidia were observed under a fluorescence microscope with an excitation light of 365 nm and an emission light of 460 nm.

[0030] Propidium iodide (PI) staining: The treated conidia were transferred to a glass slide for parallel staining. The slide was incubated with PI staining solution at 37°C for 15 min. After incubation, the staining solution was removed, and the samples were washed 2-3 times with phosphate buffer. All samples were fixed with 4% paraformaldehyde for 30 min. After covering with a coverslip and absorbing excess liquid, the slides were sealed. The stained samples were observed using a fluorescence microscope with 535 nm excitation light and 605 nm emission light.

[0031] Statistical analysis: All statistical analyses were performed using GraphPad Prism software (GraphPad software, San Diego, CA, USA). A p-value < 0.05 was considered statistically significant. Unpaired t-tests were used to analyze the survival rate of fungal colony-forming units (CFU) in different groups of untreated and nano-salt-treated strains. Statistical significance was expressed as follows: *p < 0.05; **p < 0.005; ***p < 0.001; and ****p < 0.0001.

[0032] Results and Discussion:

[0033] I. Preparation and Characterization of Sodium Chloride Nanoparticles Using an Aerosol Method: A precursor solution was atomized to generate micron-sized liquid aerosol droplets. These droplets were mixed with a heated inert carrier gas in a drying tube, causing rapid solvent evaporation. Supersaturation was achieved within the droplets, promoting the nucleation and growth of sodium chloride, ultimately forming solid nano-sized sodium chloride particles. Dynamic monitoring of particle distribution yielded sedimentation velocities and accelerations for different particle sizes. Figure 1As shown, after uniform dispersion in a sealed chamber for 30 min, 17,310 suspended nano-salt particles with a diameter range of 0.4–0.5 μm were observed, accounting for 62.7% of the total number of suspended nano-salt particles at the peak point. Particle size distribution analysis indicated that the atomized nano-salt particles were polydisperse, but still dominated by a specific fraction. Throughout the observation period, the proportion of particles in the 400–500 nm range consistently remained above 60% of the total distribution, indicating a stable and dominant subgroup. In contrast, the concentration of larger particles (500–1000 nm) gradually decreased over time. When the sampling volume was referenced, the original particle count was meaningfully interpreted; specifically, the 25,000 particles measured in the 400–1000 nm range 5 seconds after spraying corresponded to a peak number concentration of approximately 67.6 particles / cm³ in a 370 cm³ chamber. This quantification of spatial density provides a key parameter for assessing potential exposure and interaction kinetics.

[0034] In summary, the nano-salt aerosols exhibit a consistent and inhalable size distribution, with most particles concentrated in the 400-500 nm range. This well-defined physical characteristic, along with the derived particle number concentration, forms an important basis for subsequent evaluation of its environmental behavior and antifungal efficacy.

[0035] II. To investigate the antibacterial effect of nano-salt on Aspergillus fumigatus, we first selected the clinically azole-sensitive reference strain A1161. The conidia of A1161 in the untreated group and after 5 seconds of nano-salt treatment followed by a 2-hour rest in a closed chamber were counted using the plate-spreading method. Plate counts of A1161 conidia from different treatments showed that the number of conidia in the 500, 1000, and 2000 conidia / mL nano-salt treatment groups was sharply reduced compared to the untreated group. Figure 2 Survival rates for A and 2B were only 11.0%, 8.8%, and 7.2%, respectively. Figure 2 C). Furthermore, we evaluated the effects of nano-salts on environmentally sensitive and azole-resistant strains (C). Figure 2 (D and 2E). The results also demonstrated the killing effect of nano-salt on these two strains, reducing the survival rate to 9.2% and 12.5%, respectively. Figure 2 (F), indicating that the nano-salt has a significant non-selective fungicidal effect on Aspergillus fumigatus conidia.

[0036] III. Nano-salts exhibit considerable fungicidal activity against *Aspergillus flavus* and *Aspergillus niger*. Besides the most common *Aspergillus fumigatus*, other pathogenic bacteria such as *Aspergillus flavus* and *Aspergillus niger* also pose a significant clinical risk of infection. To determine whether the synthesized nano-salt particles have antibacterial efficacy against these non-*Aspergillus fumigatus* species, we intentionally selected *Aspergillus flavus* and *Aspergillus niger*, which are frequently found in clinical practice. Following the same testing procedure as for *Aspergillus fumigatus*, conidia of *Aspergillus flavus* and *Aspergillus niger* were collected, quantified, and adjusted to a concentration of 2000 CFU / mL. The suspension was then nebulized into a sealed container using a nebulizer and co-exposed with the nano-salt particles for 5 s. After exposure and the subsequent settling period, samples were collected, coated, and quantitatively counted for colonies. Therefore, we evaluated the effects of the nano-salts on these two species. Figure 3 As shown in Figure A, nano-salt treatment significantly reduced the CFU of Aspergillus flavus and Aspergillus niger. Quantitatively, the survival rates of Aspergillus flavus and Aspergillus niger were 4.3% and 20.4%, respectively. Figure 3 Consistent results were observed in independent experimental replicates (BC). Statistical comparisons showed that the survival rate of *Aspergillus niger* was significantly higher than that of *Aspergillus flavus* (p<0.01), indicating that *Aspergillus niger* possesses stronger innate resistance to nanosalts. This difference in tolerance may be attributed to the unique physiological and structural characteristics of *Aspergillus niger*, including its thick and reinforced cell wall, a robust system for maintaining internal homeostasis, and a protective melanin layer present in the spores. Compared to *Aspergillus flavus*, these characteristics may enhance its ability to resist physical damage and potential chemical oxidative stress induced by nanosalts. Therefore, under the same conditions, nanosalts showed a slightly lower killing effect on *Aspergillus niger*.

[0037] IV. Nanosalts also exhibit antifungal activity against yeast-type human fungal pathogens. It is noteworthy that, besides Aspergillus species, common environmental substrates such as air and soil are also reservoirs for other important fungal pathogens. This includes pathogenic yeasts existing in single-celled form, such as Candida albicans and Cryptococcus neoformans. According to the WHO's first list of key fungal pathogens published in 2022, Candida albicans and Candida neoformans are listed as key priority fungi posing significant public health risks. Cryptococcus neoformans typically infects humans through inhalation of environmental spores via the respiratory tract, which is the primary route of transmission from the environment to humans. On the other hand, Cryptococcus neoformans can cross the blood-brain barrier, targeting the central nervous system to cause cryptococcal meningitis, usually with a latent infection. In contrast, Candida albicans infection is primarily due to the disruption of the endogenous flora, rather than airborne transmission between individuals. Lung involvement usually occurs through endogenous inhalation rather than external transmission. Therefore, although it may enter through the respiratory tract, it is not classified as a "respiratory transmission" pathogen. Both Candida albicans and Candida neoformans are opportunistic pathogens present in the environment. Candida albicans can cause systemic infections via the bloodstream, as well as vaginal and mucosal infections, posing a significant health threat. Therefore, we further evaluated the antibacterial efficacy of the prepared nanosalt against these two fungal pathogens. Figure 4 As shown, the efficacy of nano-salts decreased with increasing yeast concentration. However, even at a concentration of 2000 CFU / mL, the survival rates of *Candida albicans* and *Candida neonicotinoids* fell below 30%. At lower concentrations (500 CFU / mL), the effect was particularly pronounced, with survival rates of 4.0% and 13.3%, respectively. Figure 4 C and 4F).

[0038] V. Nano-salts can disrupt the integrity of conidial cell walls and membranes. Given the significant antifungal effects of nano-salts on various fungi, this study used the standard strain *Aspergillus fumigatus* A1161 as a representative to investigate the effects of nano-salt treatment on the cell morphology of *Aspergillus fumigatus*. To study the killing effect of nano-salts on *Aspergillus fumigatus*, we observed the morphological changes of conidia before and after nano-salt treatment using scanning electron microscopy. Fresh spores of strain A1161 were harvested from YAG plates, then filtered and counted to prepare a spore suspension. Figure 5 As shown, untreated Aspergillus fumigatus conidia are plump spherical or nearly spherical with intact structure and fine surface ornamentation. In contrast, nano-salt treated conidia exhibit obvious structural damage and irregular morphology.

[0039] After observing the morphological changes induced by nanosalt, CFW staining was used to assess the corresponding changes in the cell walls of *Aspergillus fumigatus* in different treatment groups. CFW is a polysaccharide-specific fluorescent dye that preferentially binds to β-1,4 and β-1,3 polysaccharide chains (such as cellulose and chitin). Upon binding and excitation by ultraviolet or blue light, it emits strong blue fluorescence, thus labeling these polysaccharide structures. It is mainly used to label the cell walls of fungi and plants. Before nanosalt exposure, untreated conidia appeared as uniform, rough-surfaced spherical particles emitting bright blue-white fluorescence, mainly localized to the spore cell wall, reflecting the strong binding of CFW to chitin and cellulose in the fungal cell wall. However, in nanosalt-treated conidia, the blue-white fluorescence was concentrated inside the conidia, indicating severe damage to the cell wall. Figure 6 A). Simultaneously, Aspergillus fumigatus conidia treated with different concentrations of nano-salt were selected, and the changes in their overall fluorescence intensity were calculated using ImageJ. For example... Figure 6 As shown in Figure B, the fluorescence intensity distribution of untreated conidia exhibits a bimodal distribution. The fluorescence spectra of nano-salt-treated conidia display different trapezoidal shapes. This morphological change in fluorescence spectrum is attributed to nano-salt-induced disruption of the conidial cell wall, which facilitates CFW dye entry into the cell, leading to an increase in overall fluorescence intensity. In CFW staining, disruption of the cell wall of nano-salt-treated Aspergillus fumigatus spores was observed, and substances from outside the cell wall were detected within the cells.

[0040] Because exposure to nanosalts impairs the integrity of Aspergillus fumigatus spore cell walls, their viability was subsequently assessed. Population-level viability was assessed using propidium iodide staining. The PI dye cannot penetrate the intact membrane of living cells, only entering dead cells, where it binds to DNA and fluoresces. The intensity of the fluorescence is proportional to the DNA content. Simultaneously, PI staining further validated the viability of Aspergillus fumigatus conidia after nanosalt treatment. Figure 7 As shown in Figure A, the untreated conidia remained largely viable, unstained, and appeared as dark hollow spherical shadows against a black background, without emitting red fluorescence. Only the Aspergillus fumigatus conidia treated with nanosalt exhibited uniform bright red or orange-red fluorescence, confirming the loss of cell membrane integrity, allowing PI dye to enter, bind to nucleic acids, and produce fluorescence. Figure 7 (B) Based on these findings, the nano-salt particles primarily kill Aspergillus fumigatus by disrupting the cell wall and cell membrane, causing intracellular contents to leak out, severely damaging spore integrity, and ultimately leading to death. However, with increasing conidial concentration, the PI staining positivity rate decreased, due to reduced staining and spore aggregation detection efficiency.

[0041] In summary, the core of this study lies in the successful design and fabrication of sodium chloride nanoparticles based on antisolvent precipitation and high-energy nanotechnology, providing an innovative solution to the bottleneck in the application of water-soluble inorganic salts in aerosol disinfection. Systematic evaluation of their antifungal effects confirmed that these nanoparticles exhibit significant disinfection activity against key clinical fungal pathogens such as Aspergillus, Candida albicans, and Candida neonicotinoids.

[0042] Notably, this study elucidated the lethal mechanism of *Aspergillus fumigatus*, the main pathogen of invasive aspergillosis, at the ultrastructural level. Statistical results indicate that the nanosalt particles function by physically disrupting the structural integrity of the cell wall and cell membrane, inducing the formation of specific pores and pits, ultimately leading to irreversible leakage of intracellular contents and cell lysis. This non-selective physical damage mechanism holds promise for circumventing the bioresistance challenge faced by existing antifungal drugs and provides a promising technological approach for controlling infections caused by airborne fungi.

Claims

1. A nano-salt particle disinfectant for inhibiting pathogenic fungi, characterized in that: The process involves nano-salt particles with a particle size distribution between 400 and 1000 nm. By atomizing a 0.1 M sodium chloride aqueous solution into aerosol droplets and introducing them into a reactor, the reaction temperature is controlled between 20°C and 30°C to obtain a nano-salt particle disinfectant for inhibiting pathogenic fungi.

2. The nano-salt particle disinfectant for inhibiting pathogenic fungi according to claim 1, characterized in that: The pathogenic fungi include Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger, Candida albicans, and Cryptococcus neoformans.

3. Use of a nano-salt particle disinfectant for inhibiting pathogenic fungi, wherein the nano-salt particle disinfectant according to any one of claims 1-2 is used for the inhibition of pathogenic fungi.

4. The application of a nano-salt particle disinfectant for inhibiting pathogenic fungi according to any one of claims 1-3 against pathogenic fungi, characterized in that: The nano-salt particle disinfectant is uniformly sprayed in the disinfection space to achieve a number concentration of 20-400 nano-salt particles with a particle size of 400-1000nm in the disinfection space.

5. The application of the nano-salt particle disinfectant for inhibiting pathogenic fungi according to claim 4 against pathogenic fungi, characterized in that: The nano-salt particles are uniformly sprayed using an electric powder sprayer.

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