Synergistic mildew-proof composition and application thereof, and method for inhibiting mildew of cigar tobacco leaves

Abstract

The invention belongs to the field of antifungal agents, and particularly relates to a synergistic mildew-proof composition, application thereof and a method for inhibiting cigar tobacco leaf mildew. The synergistic mildew-proof composition is composed of phenethyl alcohol and isoamyl alcohol, and the volume ratio of phenethyl alcohol to isoamyl alcohol is (10-0.5): 1. After phenethyl alcohol and isoamyl alcohol are compounded, a strong synergistic effect is achieved, and it is guaranteed that a good anti-mildew effect can be achieved under the condition that the dosage is low. The synergistic mildew-proof composition is low in mildew-proof effective concentration and is a natural aroma substance, the potential influence of the mildew-proof process on the original flavor and sensory quality of the cigar tobacco leaves is greatly reduced, and meanwhile, the problem of residues caused by traditional chemical preservatives is also avoided.

Description

Technical Field

[0001] This invention belongs to the field of antifungal agents, specifically relating to a synergistic antifungal composition and its application, and a method for inhibiting mold growth in cigar tobacco leaves. Background Technology

[0002] Cigars undergo a lengthy process of drying, fermentation, and storage during production and processing, during which they are susceptible to infection by various molds, especially Aspergillus montmorillonite (Monticollis). Aspergillus montevidensis Drought-resistant molds such as ( ) seriously affect the quality and economic value of tobacco leaves.

[0003] Currently, anti-mold measures for cigar tobacco leaves mainly rely on strict environmental control or the effective application of chemical agents. However, the former is costly and difficult, while the latter has problems such as residues and affecting flavor. Although there are reports of inhibiting mold growth in cigar tobacco leaves through biological control, these mainly focus on the effects of single strains under specific conditions, and there is a lack of efficient and safe solutions that can be implemented throughout the entire lifecycle of cigar tobacco leaves, from dynamic drying to storage.

[0004] Chinese invention patent application CN 109370947 A, published on February 22, 2019, discloses an Enterobacterium and its application in preventing and controlling Aspergillus flavus and toxins in peanuts during storage. The volatile metabolites of this Enterobacterium, 1-pentanol and phenylethanol, have been shown to effectively inhibit the occurrence of Aspergillus flavus disease and aflatoxin contamination in peanuts.

[0005] Besides 1-pentanol and phenylethyl alcohol, monomers such as acetic acid and phenylacetaldehyde have been proven to be effective flavoring and antibacterial agents. However, when used alone, the concentration required to achieve an effective anti-mold effect is relatively high, and it may have an adverse effect on the flavor and sensory quality of cigar tobacco leaves. Summary of the Invention

[0006] The purpose of this invention is to provide a synergistic anti-mold composition that solves the problem that the high effective concentration of a single antibacterial substance can easily have an adverse effect on the flavor and sensory quality of cigar tobacco.

[0007] A second objective of the present invention is to provide the application of the above-mentioned synergistic anti-mold composition in inhibiting mold growth in cigar tobacco leaves, in order to solve the above-mentioned problems.

[0008] The third objective of this invention is to provide a method for inhibiting mold growth in cigar tobacco leaves, thereby addressing the lack of an efficient and safe solution in the prior art that can be implemented throughout the entire lifecycle of cigar tobacco leaves, from dynamic drying to storage.

[0009] To achieve the above objectives, the technical solution of the synergistic antifungal composition of the present invention is as follows: A synergistic antifungal composition comprising phenethyl alcohol and isoamyl alcohol, wherein the volume ratio of phenethyl alcohol to isoamyl alcohol is (10-0.5):1.

[0010] This invention is groundbreaking. The combination of phenylethanol and isoamyl alcohol exhibits a strong synergistic effect, ensuring excellent anti-mold performance even at low dosages. The aforementioned synergistic anti-mold composition has a low effective concentration for mold prevention and, being a natural aroma compound, significantly reduces the potential impact of the anti-mold process on the original flavor and sensory quality of cigar tobacco leaves, while also avoiding the residue problems associated with traditional chemical preservatives.

[0011] Preferably, the volume ratio of phenylethanol to isoamyl alcohol is (3.5-1.1):1.

[0012] More preferably, the volume ratio of phenylethanol to isoamyl alcohol is (2.5-1.1):1.

[0013] The application of the above-mentioned synergistic anti-mold composition in inhibiting mold growth in cigar tobacco leaves.

[0014] When the above-mentioned synergistic anti-mold composition is applied to prevent mold growth in cigar tobacco leaves, it is used at a low concentration, has little impact on the flavor and sensory quality of cigar tobacco leaves, and can be used during the drying, storage, aging and maintenance of cigar tobacco leaves, meeting the anti-mold requirements of cigar tobacco leaves throughout the entire life cycle.

[0015] The technical solution of the method for inhibiting mold growth in cigar tobacco leaves according to the present invention is as follows: A method for inhibiting mold growth on cigar tobacco leaves includes the step of applying an effective amount of the above-described synergistic anti-mold composition to the cigar tobacco leaves or their environment.

[0016] The method for inhibiting mold growth in cigar tobacco leaves of the present invention utilizes the synergistic effect of phenylethanol and isoamyl alcohol to achieve highly efficient mold prevention. It has a low application concentration, good mold prevention effect, and is suitable for drying and storage scenarios. It has good application prospects in the large-scale processing of cigar tobacco leaves.

[0017] Preferably, the application concentration of the synergistic antifungal composition is such that the final concentration of the active ingredient on the surface of the tobacco leaf is no higher than 1 / 5 of the minimum inhibitory concentration (MIC) of phenylethyl alcohol and isoamyl alcohol when used alone. The MIC is the concentration at which the inhibition rate against Aspergillus montana reaches 100% when used alone.

[0018] Preferably, the synergistic antifungal composition is applied by fumigation; when fumigating, the fumigation volume concentration of phenylethanol is 0.033-0.7 μL / mL, and the fumigation volume concentration of isoamyl alcohol is 0.022-0.3 μL / mL.

[0019] Preferably, the synergistic antifungal composition is applied by spraying, with the volume ratio of phenethyl alcohol to isoamyl alcohol controlled at (3.5-1.1):1. When spraying, 20-30 mL of the mixed reagent can be sprayed onto each tobacco stalk. The mixed reagent consists of phenethyl alcohol and isoamyl alcohol in a volume ratio of (3.5-1.1):1. Each tobacco stalk contains 50-60 tobacco leaves.

[0020] Preferably, the synergistic antifungal composition is applied during the drying, storage, or aging stages of cigar tobacco leaves.

[0021] Preferably, the synergistic anti-mildew composition is applied during cigarette maintenance.

[0022] The method for inhibiting mold growth in cigar tobacco leaves according to the present invention achieves the following beneficial effects: (1) Synergistic effect, concentration reduced by more than half: The checkerboard method experiment results show that the FIC of phenethyl alcohol and isoamyl alcohol against Aspergillus montana reaches 0.375, showing a strong synergistic effect. This means that when the same or even better antibacterial effect is achieved, the total concentration of the above synergistic antifungal composition is much lower than the concentration required for either component to be used alone.

[0023] (2) Full-cycle anti-mold, with a wide range of applications: The synergistic anti-mold composition of the present invention has been successfully applied to the drying and storage periods of cigar tobacco leaves, and has shown excellent anti-mold effects, achieving full-cycle coverage of key processing links of cigar tobacco leaves.

[0024] (3) Safety: Due to its extremely low effective concentration and the fact that the components are natural aroma substances, it greatly reduces the potential impact on the original flavor and sensory quality of cigar tobacco leaves, and avoids the residue problems caused by traditional chemical preservatives.

[0025] (4) Stable effect and strong universality: Verification experiments conducted on cigar tobacco leaves from different producing areas (Hunan, Hubei and Sichuan) all showed good anti-mold effect, indicating that the synergistic anti-mold composition and the method for inhibiting mold growth of cigar tobacco leaves of the present invention have good universality. Attached Figure Description

[0026] Figure 1 The results of MIC value determination of the inhibitory effect of phenylethanol on Aspergillus montana in the embodiments of the present invention; Figure 2 The results of MIC value determination of the inhibitory effect of isoamyl alcohol on Aspergillus montmorillonite in the embodiments of the present invention; Figure 3 The figure shows the statistical results of the inhibitory effect of the synergistic anti-mold composition on the mold growth of cigar tobacco leaves (Enshi, Hubei) during the drying process in the experimental examples of this invention. Different letters in the figure indicate significant differences between treatments. P <0.05; Figure 4The figure shows the statistical results of mold content in cigar tobacco leaves (Hunan) during storage after fumigation with different concentrations of synergistic anti-mold compositions. "Indicates a highly significant difference between the different treatments" P <0.01; Figure 5 The figure shows the statistical results of mold content in cigar tobacco leaves (Sichuan) during storage after fumigation treatment with different concentrations of synergistic anti-mold compositions in a desiccator. "Indicates that there are extremely significant differences between different treatments" P <0.0001; Figure 6 The figure shows the statistical results of mold content in cigar tobacco leaves (Sichuan) during storage after fumigation with different concentrations of synergistic anti-mold compositions within small plastic frames. "Indicates that there are extremely significant differences between different treatments" P <0.0001. Detailed Implementation

[0027] (I) A preferred embodiment of the synergistic anti-mold composition of the present invention, the application of the synergistic anti-mold composition in inhibiting mold growth in cigar tobacco leaves, and the method for inhibiting mold growth in cigar tobacco leaves.

[0028] This invention uses metabolites of a certain biocontrol strain to analyze components and identify antibacterial components such as phenylethanol, isoamyl alcohol, phenylacetaldehyde, isoamyl acetate, isobutanol, and acetic acid. Further investigation into the interactions of different antibacterial components and verification through a checkerboard method experiment revealed that phenylethanol and isoamyl alcohol inhibit *Aspergillus montana* (…). Aspergillus montevidensis It has a synergistic effect on drought-resistant molds such as )

[0029] This invention further verifies the anti-mold properties of the above-mentioned composition during the drying and storage of cigar tobacco leaves, confirming that the composition can achieve excellent anti-mold effects when used at low concentrations (e.g., not exceeding 1 / 5 of the minimum inhibitory concentration when phenylethanol and isoamyl alcohol are used alone). This creates favorable conditions for reducing the adverse effects of anti-mold measures on the quality and flavor of cigar tobacco leaves.

[0030] Compared to spraying, fumigation is a crucial method for the safe and efficient industrial processing of cigar tobacco leaves due to the minimal impact of the reagents on the leaves. This invention, through experiments using different fumigation methods such as drying room fumigation, square dish fumigation, desiccator fumigation, and plastic frame fumigation, demonstrates that the above-mentioned composition exhibits excellent anti-mold efficiency at its respective 1 / 5 MIC concentration, thus fully proving the industrial adaptability of the above-mentioned composition in fumigation-based mold prevention.

[0031] In addition, verification using cigar tobacco leaves from different origins (Hunan, Hubei, and Sichuan) proved that the above composition has a stable and universally applicable effect in preventing mold growth in cigar tobacco leaves.

[0032] The preferred embodiments described above are illustrated below with specific examples. Unless otherwise specified, the raw materials involved in the following embodiments are all commercially available products, and the relevant processing techniques are all conventional technologies.

[0033] The procedure for determining the synergistic inhibitory effect of phenylethanol and isoamyl alcohol on Aspergillus montana is as follows: (1) Activation of strain and preparation of spore suspension: Aspergillus montana seeds were inoculated into PDA solid medium and cultured at 28°C for 7 days. The spores were washed away with sterile PBS buffer to prepare a certain concentration of Aspergillus montana spore seed solution.

[0034] (2) Determination of MIC values ​​individually: The double-plate inverted method was used. A concentration gradient of phenylethanol and isoamyl alcohol was set up. The test reagent was dropped onto a filter paper disc, which was then inverted and sealed with a PDA plate inoculated with Aspergillus montana. After incubation at 28°C for 48 h, the colony diameter was measured and the inhibition rate was calculated. The concentrations of phenylethanol included: 0.2, 0.5, 1.0, 2.5, 2.8, and 2.9 μL / mL; the concentrations of isoamyl alcohol included: 0.2, 0.4, 0.6, 0.8, 0.9, and 1.0 μL / mL.

[0035] The MIC values ​​of the inhibitory effects of phenylethanol and isoamyl alcohol on Aspergillus montana are as follows: Figure 1 and Figure 2 As shown.

[0036] Figures 1-2 The results showed that the MIC of phenylethanol against Aspergillus montana was 2.8 μL / mL, and the MIC of isoamyl alcohol against Aspergillus montana was 0.9 μL / mL. Here, μL / mL represents the volume concentration, i.e., the volume of reagent used per mL of space.

[0037] (3) Synergistic effect determination: The checkerboard method was used. Phenylephrine and isoamyl alcohol were set to 2 MIC, 1 MIC, 1 / 2 MIC, 1 / 4 MIC, 1 / 8 MIC and 1 / 16 MIC, and then paired for double-plate incubation experiments. The mixtures were incubated at 28℃ for 48 h, and the inhibition rate of each combination was calculated after incubation. The results are shown in Table 1.

[0038] Table 1. Inhibition rate of different concentrations of phenylethanol and isoamyl alcohol against Aspergillus montana (checkerboard assay results) Table 1 shows that when the concentration of phenylethanol is 1 / 8 MIC (0.35 μL / mL) and the concentration of isoamyl alcohol is 1 / 4 MIC (0.225 μL / mL), the inhibition rate against Aspergillus montana remains 100%. According to the formula FCI = (0.35 / 2.8) + (0.225 / 0.9) = 0.375, FIC < 0.5, which fully demonstrates the significant synergistic inhibitory effect of phenylethanol and isoamyl alcohol on Aspergillus montana.

[0039] Furthermore, based on the data trends in Table 1, when the concentration of phenylethanol is 1 / 8 MIC and the concentration of isoamyl alcohol is 1 / 3 MIC, the result of FIC < 0.5 still exists. Similarly, when the concentrations of phenylethanol are 1 / 5 MIC, 1 / 6 MIC, and 1 / 7 MIC, and the concentration of isoamyl alcohol is 1 / 4 MIC, the result of FIC < 0.5 still exists. Based on these results, a significant synergistic effect is observed when the volume ratio of phenylethanol to isoamyl alcohol is at least in the range of (3.1-1.1):1.

[0040] Example 1 The synergistic antifungal composition of this embodiment consists of phenethyl alcohol and isoamyl alcohol, with phenethyl alcohol having a MIC concentration of 1 / 8 and isoamyl alcohol having a MIC concentration of 1 / 4. The volume ratio of phenethyl alcohol to isoamyl alcohol is 0.35:0.225 = 1.56:1.

[0041] Example 2 The synergistic antifungal composition of this embodiment consists of phenethyl alcohol and isoamyl alcohol, with phenethyl alcohol and isoamyl alcohol having a concentration of 1 / 5 of the MIC, and the volume ratio of phenethyl alcohol to isoamyl alcohol being 0.56:0.18 = 3.1:1.

[0042] Example 3 The synergistic antifungal composition of this embodiment consists of phenethyl alcohol and isoamyl alcohol, with phenethyl alcohol having a MIC concentration of 1 / 8 and isoamyl alcohol having a MIC concentration of 1 / 3. The volume ratio of phenethyl alcohol to isoamyl alcohol is 0.35:0.3 = 1.17:1.

[0043] Example 4 The synergistic antifungal composition of this embodiment consists of phenylethanol and isoamyl alcohol, with a volume ratio of phenylethanol to isoamyl alcohol of 6:1.

[0044] Example 5 The synergistic antifungal composition of this embodiment consists of phenylethanol and isoamyl alcohol, with a volume ratio of phenylethanol to isoamyl alcohol of 10:1.

[0045] In Examples 4 and 5, the relative amounts of phenylethanol and isoamyl alcohol were increased based on that in Example 2.

[0046] Example 6 The synergistic antifungal composition of this embodiment consists of phenylethanol and isoamyl alcohol, with a volume ratio of phenylethanol to isoamyl alcohol of 0.7:1.

[0047] Example 7 The synergistic antifungal composition of this embodiment consists of phenylethanol and isoamyl alcohol, with a volume ratio of phenylethanol to isoamyl alcohol of 0.5:1.

[0048] In Examples 6 and 7, the relative amounts of phenylethanol and isoamyl alcohol were increased based on that in Example 3, with the amount of isoamyl alcohol further increased.

[0049] Example 8 The application of the synergistic anti-mold composition in inhibiting mold growth in cigar tobacco leaves in this embodiment involves spraying a mixture of phenylethanol and isoamyl alcohol onto the floor of the drying room during the cigar drying stage. The concentration of phenylethanol in the drying room is controlled at 0.033 μL / mL, and the concentration of isoamyl alcohol is controlled at 0.022 μL / mL. After sealing for 48 hours, ventilation is carried out to achieve anti-mold treatment of cigar tobacco leaves during the drying stage.

[0050] Example 9 The method for inhibiting mold growth in cigar tobacco leaves in this embodiment involves fumigating the cigar tobacco leaves during storage with a combination of phenylethanol and isoamyl alcohol, wherein the volume ratio of phenylethanol to isoamyl alcohol is 3.1:1. The fumigation concentration of phenylethanol is 0.56 μL / mL, and the fumigation concentration of isoamyl alcohol is 0.18 μL / mL.

[0051] (II) Experimental Examples Example 1: Application in the drying stage of cigar tobacco leaves in Enshi, Hubei Province This experimental example examines the application effect of the above-mentioned synergistic anti-mildew composition in the drying stage of cigar tobacco leaves in Enshi, Hubei Province. The specific experimental procedure is as follows: 1. Fumigation in the drying room Mix phenylethanol and isoamyl alcohol at a volume ratio of 3:2 (3.9L: 2.6L), and spray the mixture onto the floor of the drying room (drying room volume: 120 m³). 3At this point, the concentration of phenylethanol in the drying room was 0.033 μL / mL, and the concentration of isoamyl alcohol was 0.022 μL / mL. After sealing for 48 hours, ventilation was carried out, followed by routine drying.

[0052] 2. Spraying the surface of cigar tobacco leaves During the drying and browning period (when the ambient humidity is continuously greater than 90%), a mixture of phenylethanol and isoamyl alcohol in a volume ratio of 3:2 is directly sprayed onto the surface of the tobacco leaves, with 30 mL sprayed per tobacco leaf (60 leaves in total).

[0053] The standard drying process is used for the CK group.

[0054] After drying, samples were taken and pulverized, and a bacterial suspension was obtained by shaking with sterile PBS buffer. After serial dilution, the suspension was spread onto Bengal red solid medium and incubated at 28°C for 5 days. The number of mold colonies under different treatments was counted. Specific results are shown below. Figure 3 As shown.

[0055] Depend on Figure 3 It can be seen that, compared with the control group, both fumigation and surface spraying treatments significantly inhibited mold growth in tobacco leaves. Mold counting results showed that the mold count in the treated group was significantly lower than that in the control group, proving that the above-mentioned composition has excellent anti-mold effects in a dynamically humid drying environment.

[0056] Experimental Example 2: Application of Anti-mold Measures in Multi-origin and Multi-method Storage Stages This experimental example verifies the anti-mold effect of the above-mentioned synergistic anti-mold composition during storage by using multiple production sites and methods.

[0057] 1. Inverted square dish method (Hunan cigar tobacco) For Hunan cigar tobacco leaves, the anti-mold effect of the composition during storage was tested using the square dish inverted method. The specific experimental steps are as follows: ① Tobacco leaf cutting: Cut the cigar tobacco leaves into 2 cm × 2 cm pieces and rehydrate them until the moisture content is about 30%. Select 40 cigar tobacco leaves and place them in a square dish.

[0058] ② Concentration settings: Three concentration settings are provided: 1 / 5 MIC, 1 / 10 MIC, and 1 / 20 MIC. This means the concentrations of phenylethanol and isoamyl alcohol are 1 / 5, 1 / 10, and 1 / 20 of their respective minimum inhibitory concentrations. Add the above concentrations of phenylethanol and isoamyl alcohol dropwise onto filter paper, control the concentration, and seal two square dishes by inverting them together. Incubate at 28℃ for 5 days.

[0059] ③ Mold count: After the culture was completed, the number of leaves with mold in cigar tobacco under different treatments was counted, and the mold content under different treatments was determined by Bengal red medium.

[0060] Table 2 shows the statistical results of mold rate in Hunan cigar tobacco leaves under different treatments.

[0061] Table 2. Statistics on mold rates of Hunan cigar tobacco leaves after fumigation with different concentrations of synergistic compositions using a square dish-to-plate method. As shown in Table 2, all tobacco leaves in the CK treatment group were moldy, with a mold rate of 100%. At a concentration of 1 / 5 MIC, the mold rate was 75%, and at a concentration of 1 / 20 MIC, the mold growth was still inhibited in cigar tobacco leaves.

[0062] The statistical results of mold content are as follows: Figure 4 As shown. By Figure 4 It can be seen that the mold content under different concentrations of the composition treatment was significantly different from that of the control.

[0063] 2. Dryer fumigation method (Sichuan Dexue cigar tobacco leaves) For Sichuan Dexue cigar tobacco leaves, the anti-mold effect of the composition during storage was tested using a desiccator fumigation method. The specific experimental steps are as follows: ① Tobacco leaf cutting: Cut the cigar tobacco leaves into 2 cm × 2 cm pieces and rehydrate them until the moisture content is about 30%. Select 100 cigar tobacco leaves and place them in a desiccator.

[0064] ② Add different concentrations of the compound reagent to the bottom of the desiccator. The concentration settings include 1 / 2 MIC and 1 / 4 MIC; 1 / 2 MIC means that the concentrations of phenylethanol and isoamyl alcohol are both half of their respective minimum inhibitory concentrations; the meaning of 1 / 4 MIC is similar. After sealing, incubate at 28℃ for 5 days.

[0065] ③ Mold count: After the culture was completed, the number of leaves with mold in cigar tobacco under different treatments was counted, and the mold content under different treatments was determined by Bengal red medium.

[0066] The results of mold growth rate of cigar tobacco leaves under different composition concentrations are shown in Table 3.

[0067] Table 3. Statistics on mold rate of Sichuan Dexue cigar tobacco leaves after fumigation with different concentrations of synergistic compositions using the dryer method. As shown in Table 3, the mold rate of cigar tobacco leaves was 0% under both 1 / 2 MIC and 1 / 4 MIC treatments, while the mold rate of cigar tobacco leaves was 93% under the CK treatment.

[0068] The statistical results of mold content are as follows: Figure 5 As shown.

[0069] Figure 5The results showed that the mold content at 1 / 2 MIC and 1 / 4 MIC concentrations was significantly different from that in the control group, thus proving that the above composition had a very significant effect on inhibiting mold growth in Sichuan cigar tobacco leaves.

[0070] 3. Fumigation method using small plastic frames (Sichuan Dexue cigar tobacco leaves) For Sichuan Dexue cigar tobacco leaves, the anti-mold effect of the composition during storage was tested using the plastic frame fumigation method. The specific experimental steps are as follows: ① Tobacco leaf cutting: Cut the cigar tobacco leaves into 2 cm × 2 cm pieces and rehydrate them until the moisture content is about 30%. Select 100 cigar tobacco leaves and place them in a small plastic box.

[0071] ② Add different concentrations of the composition dropwise onto filter paper discs, place the filter paper discs under a small plastic frame, and then seal the plastic frame with a resealable bag. The composition concentrations are set as follows: 1 / 5 MIC, 1 / 10 MIC, and 1 / 20 MIC. 1 / 5 MIC means that the concentrations of phenylethanol and isoamyl alcohol are both 1 / 5 of their respective minimum inhibitory concentrations (MICs), and the meanings of 1 / 10 MIC and 1 / 20 MIC are similar. After sealing, incubate at 28°C for 5 days.

[0072] ③ Mold count: After the culture was completed, the number of leaves with mold in cigar tobacco under different treatments was counted, and the mold content under different treatments was determined by Bengal red medium.

[0073] The results of mold growth rate of cigar tobacco leaves under different composition concentrations are shown in Table 4.

[0074] Table 4. Statistical results of mold growth rate of Sichuan Dexue cigar tobacco leaves after fumigation with different concentrations of synergistic compositions in plastic frames. As shown in Table 4, the mold rate of Sichuan Dexue cigar tobacco leaves was 0% at a 1 / 5 MIC concentration, 38% at a 1 / 10 MIC concentration, and 73% at a 1 / 20 MIC concentration.

[0075] The results of mold count under different composition concentrations are as follows: Figure 6 As shown.

[0076] Figure 6 The results showed that the mold content in all three concentration treatments was significantly different from that in the control (CK) treatment. This indicates that the above-mentioned synergistic antifungal composition has a stable antifungal effect.

[0077] Interactions under other combinations of antibacterial substances in Experiment Example 3 The interaction effects of antibacterial agents such as phenylethanol, isoamyl alcohol, phenylacetaldehyde, isoamyl acetate, isobutanol, and acetic acid on Aspergillus montana were investigated using the checkerboard method, exploring the interactions between different substances. Specifically, Aspergillus montana was first used as the research object, and the minimum inhibitory concentration (MIC) of Aspergillus montana was studied with different reagents. Based on this, the interactions between pairs of substances were studied using the checkerboard method. The specific determination procedure was the same as that for determining the synergistic inhibitory effect of phenylethanol and isoamyl alcohol on Aspergillus montana. The FIC determination results are shown in Table 5.

[0078] Table 5. Results of FIC assay for different combinations of antibacterial substances against Aspergillus montana. As shown in Table 5, among the six reagents tested, only phenylethyl alcohol and isoamyl alcohol showed a significant synergistic effect; the other reagents did not exhibit synergistic effects. Specifically, the reactions between isoamyl acetate and isoamyl alcohol, isoamyl acetate and isobutanol, isoamyl acetate and phenylacetaldehyde, phenylacetaldehyde and acetic acid, phenylacetaldehyde and isoamyl alcohol, acetic acid and isoamyl alcohol, and isobutanol and phenylethyl alcohol were additive; the reactions between isoamyl acetate and phenylethyl alcohol, isoamyl acetate and acetic acid, phenylacetaldehyde and isobutanol, acetic acid and isobutanol, and acetic acid and phenylethyl alcohol were independent; and the reaction between phenylacetaldehyde and phenylethyl alcohol was antagonistic.

[0079] In conclusion, the above-mentioned experiments strongly demonstrate that the synergistic anti-mold composition of the present invention can have a significant anti-mold effect on cigar tobacco leaves from various sources, even at concentrations as low as 1 / 5 MIC, through fumigation. This large-scale industrial anti-mold treatment of cigar tobacco leaves is extremely advantageous.

[0080] Finally, it should be noted that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A synergistic antifungal composition, characterized in that, It is composed of phenylethanol and isoamyl alcohol, with a volume ratio of phenylethanol to isoamyl alcohol of (10-0.5):

1.

2. The synergistic antifungal composition as described in claim 1, characterized in that, The volume ratio of phenylethanol to isoamyl alcohol is (3.5-1.1):

1.

3. The synergistic antifungal composition as described in claim 2, characterized in that, The volume ratio of phenylethanol to isoamyl alcohol is (2.5-1.1):

1.

4. The use of a synergistic antifungal composition as described in any one of claims 1-3 in inhibiting mold growth in cigar tobacco leaves.

5. A method for inhibiting mold growth in cigar tobacco leaves, characterized in that, The step includes applying an effective amount of the synergistic antifungal composition as described in any one of claims 1-3 to cigar tobacco leaves or their environment.

6. The method for inhibiting mold growth in cigar tobacco leaves as described in claim 5, characterized in that, The application concentration of the synergistic antifungal composition is such that the final concentration of its active ingredients on the surface of tobacco leaves is no higher than 1 / 5 of the minimum inhibitory concentration of phenylethanol and isoamyl alcohol when used alone.

7. The method for inhibiting mold growth in cigar tobacco leaves as described in claim 5, characterized in that, The synergistic antifungal composition is applied by fumigation. When fumigating, the fumigation volume concentration of phenylethanol is 0.033-0.7 μL / mL, and the fumigation volume concentration of isoamyl alcohol is 0.022-0.3 μL / mL.

8. The method for inhibiting mold growth in cigar tobacco leaves as described in claim 5, characterized in that, The synergistic antifungal composition is applied by spraying. During spraying, the volume ratio of phenylethanol to isoamyl alcohol is controlled at (3.5-1.1):

1.

9. The method for inhibiting mold growth in cigar tobacco leaves as described in any one of claims 5-8, characterized in that, The synergistic antifungal composition is applied during the drying, storage, or aging stages of cigar tobacco leaves.

10. The method for inhibiting mold growth in cigar tobacco leaves as described in any one of claims 5-8, characterized in that, The synergistic anti-mildew composition is applied during cigarette maintenance.

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