Composite plant essential oil bacteriostatic composition and bacteriostatic agent thereof

Abstract

The invention relates to the technical field of natural bacteriostatic compositions, in particular to a composite plant essential oil bacteriostatic composition and a bacteriostatic agent thereof, the composite plant essential oil bacteriostatic composition comprises active ingredients and a pharmaceutically acceptable carrier, and the active ingredients comprise cinnamon essential oil, peppermint essential oil and dandelion essential oil in a mass ratio of 1: 1-2: 2-4. Essential oil extracted from natural plants is utilized, so that the growth of the mould is effectively inhibited, and meanwhile, the content of toxins generated by the mould is reduced. According to the invention, not only are environmental pollution and health risk possibly brought by a chemical antibacterial agent avoided, but also a safer and more environment-friendly alternative scheme is provided. In the fields of livestock breeding, food, medical treatment and the like, product quality can be improved, health of consumers is guaranteed, and green sustainable development is promoted.

Description

Technical Field

[0001] The present invention relates to the technical field of natural antibacterial compositions, in particular to a composite plant essential oil antibacterial composition and an antibacterial agent thereof. Background Art

[0002] Mold and mycotoxins are widely present in agricultural products, feed ingredients, and animal diets, seriously impacting animal performance and human health, and have become a problem that cannot be ignored. Currently, there are over 400 known mycotoxins, among which aflatoxin B1 (AFB1), zearalenone (ZEN), and vomitoxin (DON) are the most common mycotoxins found in feed and raw materials.

[0003] Currently, common methods for removing mycotoxins include physical detoxification, chemical detoxification, and biological detoxification. Common physical detoxification methods include adding adsorbents, removing mold particles, heat detoxification, and radiation. Adding adsorbents to feed to absorb aflatoxins is a common detoxification method in the livestock and poultry industry. Removing mold particles takes advantage of the low specific gravity of moldy particles, allowing them to be separated by air. This method is time-consuming and labor-intensive, and is suitable for treating small quantities of moldy feed. Heat detoxification involves baking contaminated corn, sorghum, and other materials at high temperatures. This method can remove approximately 60% of mycotoxins, but high-temperature treatment also reduces the nutritional value of the feed. Radiation involves spreading aflatoxin-containing feed into a thin layer and then irradiating it with ultraviolet light. This method can remove approximately 97% of the toxins, but is only suitable for laboratory processing.

[0004] Chemical detoxification involves treating moldy feed with alkali or oxidizing agents. Mycotoxins decompose and become inactivated when exposed to alkali or oxidizing agents, achieving the desired detoxification effect. Commonly used chemicals include ammonia, limewater, caustic soda, and sodium hypochlorite. Chemical detoxification can affect the nutritional value and palatability of feed. Certain chemicals are only effective against certain mycotoxins. For example, ammonia is effective against aflatoxins but less effective against other toxins. Chemical treatment also poses potential risks, including residual residues.

[0005] Biodetoxification utilizes specialized microorganisms or enzymes produced by their metabolism to degrade mycotoxins in feed. This gentle microbial degradation reduces the use of strong acids and alkalis, which can damage feed nutrients. The microbial degradation process does not introduce toxic or harmful chemicals, minimizing nutrient loss. While biodetoxification can specifically degrade some mycotoxins, enzyme activity is unstable, resulting in high costs and a high price tag. Furthermore, whether the secondary products produced after enzymatic hydrolysis are guaranteed to be non-toxic remains to be determined. Summary of the Invention

[0006] The present invention aims to overcome the shortcomings of the existing technology by providing a composite plant essential oil antibacterial composition and its antibacterial agent. This invention utilizes essential oils extracted from natural plants to effectively inhibit mold growth while reducing the toxin content produced by mold. This invention not only avoids the environmental pollution and health risks associated with chemical antibacterial agents but also provides a safer and more environmentally friendly alternative. In the fields of animal husbandry, food, and healthcare, this invention can help improve product quality, protect consumer health, and promote green and sustainable development.

[0007] The present invention is achieved through the following technical solutions: On the one hand, a composite plant essential oil antibacterial composition is provided, comprising active ingredients and a pharmaceutically acceptable carrier, wherein the active ingredients include cinnamon essential oil, peppermint essential oil, and dandelion essential oil in a mass ratio of 1:1 to 2:2 to 4.

[0008] Through the above technical solution, cinnamon essential oil has the effects of promoting blood circulation, improving poor circulation, relieving muscle spasms and rheumatism, and having antibacterial and psychological benefits. Furthermore, cinnamon essential oil's active ingredient, cinnamaldehyde, has a significant inhibitory effect on molds and toxins such as Aspergillus flavus, destroying mold hyphae, inhibiting spore formation and growth, and reducing the pathogenic effects of mycotoxins.

[0009] Peppermint essential oil, primarily composed of volatile metabolites such as menthol and menthone, possesses anti-inflammatory, antibacterial, antiviral, immunomodulatory, anti-tumor, neuroprotective, anti-fatigue, and antioxidant properties. Peppermint essential oil has significant inhibitory effects against molds such as Aspergillus flavus. Studies have shown that peppermint essential oil can disrupt the cell structure of molds, inhibiting their growth and reproduction. Peppermint essential oil can also inhibit the production of toxins, such as aflatoxin B1, produced by Aspergillus flavus and other molds. By inhibiting the synthesis of toxins, peppermint essential oil can reduce the harmful effects of mycotoxins on the body.

[0010] Dandelion essential oil has significant anti-inflammatory effects, can reduce inflammatory responses, relieve symptoms such as pain and swelling; it is rich in antioxidants, can scavenge free radicals, protect cells from oxidative damage, and delay the aging process; it has inhibitory effects on a variety of bacteria and fungi, can destroy the cell structure of pathogens, and inhibit their growth and reproduction; it stimulates the secretion of bile, promotes the metabolism of fat and toxins, and helps improve digestive function.

[0011] Furthermore, cinnamon essential oil (rich in cinnamaldehyde) and peppermint essential oil (dominated by menthol) physically disrupt the fungal cell membrane by destroying its lipid bilayer. The triterpenoids in dandelion essential oil further interfere with the activity of enzymes that synthesize the fungal cell wall. These three ingredients combine to create a dual effect of "membrane disruption and enzyme inhibition," significantly enhancing the effectiveness of killing stubborn molds like Aspergillus flavus. This combination is the first to establish a "three-in-one" antibacterial, antitoxin, and host protection system, effectively killing molds and removing toxins. Based on a mathematical model developed using an orthogonal experimental design, the optimal ratio of the three essential oils was determined, minimizing the dosage of the active ingredients.

[0012] Furthermore, the active ingredient also includes garlic essential oil and / or citronella essential oil; the mass ratio of the cinnamon essential oil, peppermint essential oil, dandelion essential oil, garlic essential oil, and citronella essential oil is 1:1-2:2-4:1-4:2-4.

[0013] Through the above technical solution, garlic essential oil is rich in multiple active ingredients. These ingredients exert powerful antibacterial effects in the body, effectively inhibiting or killing a variety of common pathogens, such as Escherichia coli and Staphylococcus aureus. Garlic oil also modulates inflammatory responses, alleviates inflammatory symptoms, and promotes the body's self-repair capabilities. Its antioxidant properties can scavenge free radicals in the body, thereby slowing cell aging and maintaining a healthy state of the body. It also activates the gastrointestinal mucosa, stimulates the secretion of digestive juices, and has a stomach-tonifying and intestinal-regulating effect, improving appetite, promoting digestion, and reducing gastrointestinal burden.

[0014] Citronella essential oil, whose main components include citronellol, citral, eugenol, and zingiberene, has inhibitory effects on a variety of fungi, including Aspergillus and Penicillium. Studies have shown that citronella essential oil can significantly reduce the growth and reproduction of fungi, effectively reducing the risk of fungal infections. Furthermore, citronella essential oil has been shown to enhance immune function, promote wound healing, and have antibacterial, anti-inflammatory, antiemetic, and antipruritic properties.

[0015] The five components combined above form a three-tiered system: membrane protein, phospholipid bilayer, and transmembrane transport. Cinnamaldehyde (cinnamon essential oil) and allicin (garlic essential oil) synergistically disrupt the fungal cell membrane structure, citronellol (citronella essential oil) anchors membrane proteases through hydrogen bonding, and menthol enhances membrane fluidity, promoting transmembrane penetration of the active ingredient. Compared to existing technologies, this combination transcends the antimicrobial limitations of traditional ternary combinations, constructing a trinity detoxification system: toxin adsorption, metabolic activation, and excretion promotion. This self-assembly structure enhances product stability. Through optimized proportions, the five components create a greater synergistic effect, maintaining system stability, effectively inhibiting fungal growth, and further safeguarding animal health.

[0016] Furthermore, the pharmaceutically acceptable carrier includes an emulsifier, which is selected from at least one of Tween 80, propylene glycol, and tristyrylphenol polyoxyethylene ether, and the added amount of the emulsifier is 2 to 4 times the total weight of the active ingredient.

[0017] The technical advantages of the emulsifier described above are: Tween 80 forms a hydrogen bond network with terpenoids in plant essential oils, improving product stability; enhancing transdermal penetration; and, when used in combination with essential oils, creating a synergistic antimicrobial effect. Malondialdehyde effectively inhibits the oxidative decomposition of essential oil components, forms a microemulsion with triterpenoids, stabilizes the system's viscosity, and maintains the emulsion's low-temperature stability through a dehydrogenation bond network. Tristyrylphenol polyoxyethylene ether exhibits high drug loading, shear stability, and long-lasting sustained-release properties. For practical production applications, tristyrylphenol polyoxyethylene ether is a preferred carrier for this product.

[0018] Furthermore, the content standards of active ingredients in the composition are: cinnamaldehyde content ≥0.02%, menthol content ≥0.02%, taraxasterol content ≥0.02%, allicin content ≥0.06%, and geraniol content ≥0.03%.

[0019] Through the above technical solution, the combination of cinnamaldehyde and allicin disrupts microbial cell membranes, while citronellal and menthol jointly inhibit fungal biofilm formation, improving mold biofilm clearance. Taraxasterol and allicin, as well as cinnamaldehyde and allicin, synergistically enhance the body's anti-inflammatory and antioxidant capabilities. The synergistic effects of these components expand the multi-target antimicrobial spectrum, optimize product stability, and enhance product safety.

[0020] A composite plant essential oil antibacterial agent is also provided, which contains the composite plant essential oil antibacterial composition.

[0021] Furthermore, the antibacterial agent has an inhibitory effect on Aspergillus flavus and Fusarium graminearum, especially on Aspergillus flavus and aflatoxins B1, B2, G1, and G2.

[0022] Through the above technical scheme, the present invention not only has a strong inhibitory effect on the growth of Aspergillus flavus through the synergistic effect of composite plant essential oils, but also can broadly degrade aflatoxins B1, B2, G1, and G2, with a degradation rate of >60%; its natural ingredients have no toxic residues, and are suitable for multiple scenarios such as grain storage, processing and pre-packaging, with both high efficiency and safety.

[0023] Beneficial effects

[0024] The present invention utilizes multiple plant essential oils, breaking through the traditional single-ingredient or simple compounding model. Through targeted complementary effects between the ingredients, it effectively inhibits mold growth while reducing the toxin content produced by the mold. At the same time, by improving product safety and stability, it addresses the volatility of traditional plant formulas. This invention not only avoids the environmental pollution and health risks that may be caused by chemical antimicrobial agents, but also provides a safer and more environmentally friendly alternative. In the fields of animal husbandry, food, and medical care, it helps improve product quality, protect consumer health, and promote green and sustainable development. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 Effects of compound plant essential oils on mycotoxin production;

[0026] Figure 2 Effects of compound plant essential oils on mycotoxin production;

[0027] Figure 3 Effects of compound plant essential oils on mycotoxin production;

[0028] Figure 4 The effect of compound plant essential oils on degrading mycotoxins in vivo;

[0029] Figure 5 Effects of different compound plant essential oils on degrading mycotoxins in vivo;

[0030] Figure 6 Determination of the effect of different compound plant essential oils on degrading aflatoxins B1, B2, G1 and G2 in vivo. DETAILED DESCRIPTION

[0031] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0032] All other embodiments obtained by persons of ordinary skill in the art based on the embodiments herein without inventive effort are intended to fall within the scope of protection of the present invention. Experimental methods in the following examples, where specific conditions are not specified, were generally performed under conventional conditions or as recommended by the manufacturer. Unless otherwise indicated, all percentages, ratios, proportions, and parts are by weight.

[0033] Unless otherwise specified, the reagents and raw materials used in the examples and comparative examples of the present invention can be obtained through commercial channels.

[0034] Example 1

[0035] Aspergillus flavus and Fusarium graminearum were inoculated on potato dextrose agar (PDA) and cultured at 28°C for 7 days. The spore suspension was eluted with physiological saline containing 0.1% Tween 80 and the concentration of the spore suspension was adjusted to 10 6 ~10 7 CFU / mL.

[0036] Test Example 1: Determination of the size of the inhibition zone of mold by a single plant essential oil

[0037] Evenly spread 100 μL of the bacterial suspension onto PDA medium. Use a microporator to create a 5 mm diameter circular well in the medium. Pipette 100 μL of 5 g / L plant essential oil into the well. Incubate at 28°C for 3–5 days. Determine the diameter of the inhibition zone using the cross-hatch method. Use distilled water as a negative control. Criteria for high sensitivity include an inhibition zone diameter >20 mm; moderate sensitivity: an inhibition zone diameter between 15–19 mm; and low sensitivity: an inhibition zone diameter between 10–14 mm.

[0038] Table 1 Antibacterial activity of plant essential oils against mold (inhibition zone, diameter mm)

[0039] essential oils Aspergillus flavus Fusarium graminearum Cinnamon essential oil 22.1±0.22 18.3±0.35 Peppermint essential oil 19.1±0.19 20.5±0.17 Dandelion essential oil 21.8±0.30 19.6±0.43

[0040] As shown in Table 1, Aspergillus flavus and Fusarium graminearum were highly sensitive to the three plant essential oils.

[0041] Test Example 2: Determination of the Minimum Inhibitory Concentration (MIC) of a Single Plant Essential Oil Against Mold

[0042] Using the two-fold dilution method, plant essential oils were diluted and quantitatively added to PDA culture medium at 50°C–55°C. After the culture medium cooled and solidified, 100 μL of the bacterial solution was evenly spread on the medium and incubated in a 28°C incubator for 5 days. The MIC of the strain was determined as the concentration of plant extract that resulted in no fungal growth observed by naked eye. Solvent was added as a negative control.

[0043] Table 2 MIC values ​​of plant essential oils against the tested bacteria (mg / mL)

[0044] essential oils Aspergillus flavus Fusarium graminearum Cinnamon essential oil 0.039 0.039 Peppermint essential oil 0.039 0.078 Dandelion essential oil 0.039 0.078

[0045] As shown in Table 2, the three essential oils all had good inhibitory effects on the test strains.

[0046] Test Example 3: Determination of the Minimum Inhibitory Concentration (MIC) of Compound Plant Essential Oils against Mold

[0047] Cinnamon oil, peppermint oil, and dandelion oil were accurately weighed and mixed in a mass ratio of 1:(1-2):(1-4). Tristyrylphenol polyoxyethylene ether, Tween 80, or propylene glycol were added and stirred thoroughly to obtain the compound essential oil. Five dilutions of the essential oils used in the antifungal test were diluted two-fold in an ELISA plate. Using an orthogonal experimental design, each solution was pipetted until each well contained 100 μL of the mixed essential oil and 100 μL of the bacterial suspension. The mixture was then incubated in a 28°C incubator for 5 days.

[0048] Table 3 Inhibitory effect of compound essential oil on mold

[0049]

[0050] Note: “+” indicates that colonies are formed in the culture medium, and “–” indicates that no colonies are formed in the culture medium.

[0051] As shown in Table 3, the mixture of cinnamon essential oil, peppermint essential oil and dandelion essential oil in a mass ratio of 1:1:2, 1:2:2 and 1:2:4 can effectively inhibit the growth of fungi.

[0052] Test Example 4: Effect of plant essential oils on mycotoxin production

[0053] Take 100 mL of potato dextrose water (PDB) medium, divide it into Erlenmeyer flasks, and sterilize it by autoclaving until ready for use. Pipette the plant essential oil dilution into PDB, add 100 μL of bacterial suspension to each Erlenmeyer flask, and incubate at 28°C for 5 days. Filter the mycelium, and determine the toxin content using immunoaffinity column-high-performance liquid chromatography.

[0054] Depend on Figure 1 It can be seen that different concentrations of composite plant essential oils can effectively inhibit the production of mycotoxins. Among them, the composite essential oil composed of cinnamon essential oil, peppermint essential oil, and dandelion essential oil in a mass ratio of 1:2:2 has the best effect.

[0055] Example 2

[0056] Test Example 1: Determination of the Minimum Inhibitory Concentration (MIC) of a Single Plant Essential Oil Against Mold

[0057] Aspergillus flavus and Fusarium graminearum were inoculated on PDA culture medium and cultured at 28°C for 7 days. Spore suspension was eluted with physiological saline containing 0.1% Tween 80, and the concentration of the spore suspension was adjusted to 106-107 CFU / mL.

[0058] Plant essential oils were diluted using the two-fold dilution method and quantitatively added to PDA culture medium at approximately 50°C. After the culture medium cooled and solidified, 100 μL of the bacterial solution was evenly spread on the medium and incubated in a 28°C incubator for 5 days. The concentration of plant extract that resulted in no fungal growth was defined as the MIC for the strain. Solvent was added as a negative control.

[0059] Table 4 MIC values ​​of plant essential oils against the tested strains (mg / mL)

[0060]

[0061]

[0062] As shown in Table 4, the four plant essential oils have a good inhibitory effect on Aspergillus flavus and Fusarium graminearum.

[0063] Test Example 2: Determination of the Minimum Inhibitory Concentration (MIC) of Compound Plant Essential Oils against Mold

[0064] Accurately weigh cinnamon essential oil, peppermint essential oil, dandelion essential oil, and garlic essential oil in a mass ratio of 1:(1-2):(2-4):(1-4), slowly stir and mix thoroughly, then add any one of tristyrylphenol polyoxyethylene ether, Tween 80, or propylene glycol and mix thoroughly to obtain a compound plant essential oil. Pipette each solution until each well contains 100 μL of the mixed essential oil and 100 μL of the bacterial suspension. Place in a 28°C incubator and incubate for 5 days.

[0065] Table 5 Inhibitory effect of compound essential oil on mold

[0066]

[0067] Note: “+” indicates that colonies are formed in the culture medium, and “–” indicates that no colonies are formed in the culture medium.

[0068] It can be seen from Table 5 that cinnamon essential oil, peppermint essential oil, dandelion essential oil, and garlic essential oil mixed in mass ratios of 1:1:2:4, 1:2:2:2, and 1:2:4:2 can effectively inhibit the growth of fungi.

[0069] Test Example 3: Effect of plant essential oils on mycotoxin production

[0070] Prepare 100 mL of PDB culture medium and dispense it into Erlenmeyer flasks. Autoclave and sterilize until ready for use. Pipette the plant essential oil solution from Experiment 2 into PDB. Add 100 μL of the bacterial suspension to each Erlenmeyer flask. Incubate at 28°C for 5 days. Filter the mycelium and determine the toxin content using an immunoaffinity column-HPLC method.

[0071] Depend on Figure 2 It can be seen that different concentrations of composite plant essential oils can effectively inhibit the production of mycotoxins.

[0072] In summary, a mixture of cinnamon, peppermint, dandelion, and garlic essential oils in a mass ratio of 1:(1-2):(2-4):(2-4) can effectively inhibit fungal growth. Among them, a mass ratio of 1:1:2:4 among the four plant essential oils showed a better effect in inhibiting mold growth and mycotoxin production.

[0073] Example 3

[0074] Test Example 1: Determination of the Minimum Inhibitory Concentration (MIC) of a Single Plant Essential Oil Against Mold

[0075] Aspergillus flavus and Fusarium graminearum were inoculated on PDA culture medium and cultured at 28°C for 7 days. Spore suspension was eluted with physiological saline containing 0.1% Tween 80, and the concentration of the spore suspension was adjusted to 106-107 CFU / mL.

[0076] Plant essential oils were diluted two-fold and quantitatively added to PDA culture medium at approximately 50°C. After the culture medium cooled and solidified, 100 μL of the bacterial solution was evenly spread over the medium and incubated in a 28°C incubator for 5 days. The concentration of plant extract that resulted in no fungal growth was defined as the MIC for the strain. Solvent was added as a negative control.

[0077] Table 6 MIC values ​​of plant essential oils against the tested strains (mg / mL)

[0078] essential oils Aspergillus flavus Fusarium graminearum Cinnamon essential oil 0.039 0.039 Peppermint essential oil 0.039 0.078 Dandelion essential oil 0.078 0.156 Garlic essential oil 0.156 0.039 Citronella essential oil 0.078 0.156

[0079] As shown in Table 6, the five plant essential oils have strong antibacterial activity against Aspergillus flavus and Fusarium graminearum.

[0080] Test Example 2: Determination of the Minimum Inhibitory Concentration (MIC) of Compound Plant Essential Oils against Mold

[0081] Accurately weigh cinnamon essential oil, peppermint essential oil, dandelion essential oil, garlic essential oil, and citronella essential oil in a mass ratio of 1:(1-2):(2-4):(1-4):(2-4), slowly stir and mix thoroughly, then add any one of tristyrylphenol polyoxyethylene ether, Tween 80, or propylene glycol and mix thoroughly to obtain a compound plant essential oil. Pipette each solution until each well contains 100 μL of the mixed essential oil and 100 μL of the bacterial suspension. Place the well in a 28°C incubator and incubate for 5 days.

[0082] Table 7 Inhibitory effect of compound plant essential oil on mold

[0083]

[0084]

[0085] Note: “+” indicates that colonies are formed in the culture medium, and “–” indicates that no colonies are formed in the culture medium.

[0086] As shown in Table 7, cinnamon essential oil, peppermint essential oil, dandelion essential oil, garlic essential oil, and citronella essential oil mixed in mass ratios of 1:2:4:4:2, 1:1:2:2:2, and 1:1:2:1:4 can effectively inhibit the growth of fungi.

[0087] Test Example 3: Determination of the active ingredients of the fungus-inhibiting composite plant essential oil

[0088] In Experimental Example 2, the active ingredients of treatment group 1, which had the best effect on mold, were determined according to the 2020 edition of the Chinese Pharmacopoeia, Part I of cinnamon essential oil, peppermint essential oil, and dandelion essential oil, and according to GB1886.272-2016 and GB1886.271-2016, respectively, for garlic essential oil and citronella essential oil, and the area normalization method was used to calculate the content of each active ingredient.

[0089] Testing revealed that the crude essential oil contained cinnamaldehyde ≥ 75%, menthol ≥ 50%, taraxasterol ≥ 20%, allicin ≥ 63%, and citronellal ≥ 60%. In treatment group 1, which demonstrated the best antibacterial effect, cinnamaldehyde ≥ 0.02%, menthol ≥ 0.02%, taraxasterol ≥ 0.02%, allicin ≥ 0.06%, and geraniol ≥ 0.03%.

[0090] Test Example 4: Effect of plant essential oils on mycotoxin production

[0091] Prepare PDB culture medium and dispense it into Erlenmeyer flasks. Sterilize with high-pressure steam before use. Pipette the plant essential oil solution from Experiment 2 into PDB, add 100 μL of the bacterial suspension, and culture at 28°C for 5 days. Filter the mycelia and determine the toxin content using immunoaffinity column-HPLC.

[0092] Depend on Figure 3 It can be seen that different concentrations of composite plant essential oils can effectively inhibit the production of mycotoxins.

[0093] Test Example 5: Stability test of compound plant essential oil

[0094] Accurately weigh cinnamon essential oil, peppermint essential oil, dandelion essential oil, garlic essential oil, and citronella essential oil in a mass ratio of 1:(1-2):(2-4):(1-4):(2-4). Slowly stir and mix thoroughly. Add 2-4 times the mass of any one of tristyrylphenol polyoxyethylene ether, Tween 80, or propylene glycol. Incubate at 25°C, 40°C, or –20°C for 30 days and observe whether the essential oil solution separates. Then, add 1-10 times the mass of distilled water or tap water and observe whether oil droplets appear.

[0095] The results showed that when 2 to 4 times the mass of the composite essential oil was added to tristyrylphenol polyoxyethylene ether, Tween 80 and propylene glycol, no stratification occurred at different temperatures. After adding water, the essential oil solution was fully dissolved and no oil droplets floated on the liquid surface.

[0096] Test Example 6: Determination of the effect of compound plant essential oil on degrading mycotoxins in vivo

[0097] 120 broiler chickens at 15 days old were fed the following diets and divided into 3 groups for 7 consecutive days.

[0098] Group 1: fed a basal diet (AFB1 ≤ 50 μg / kg, ZEA ≤ 500 μg / kg, DON ≤ 5000 μg / kg) with normal drinking water; Group 2: fed a basal diet (moldy corn) with normal drinking water; Group 3: fed a basal diet (moldy corn) with drinking water supplemented with a compound plant essential oil. On day 8, five chickens from each group were killed and their intestinal contents removed. A 1g sample was mixed with a methanol-water solution (85:15 volume ratio) to prepare a homogenate. The homogenate was centrifuged, and the upper methanol solution was removed. 10% sodium chloride was added and the solution was extracted twice with chloroform. The chloroform layer was collected, dried under reduced pressure with nitrogen, dissolved in acetonitrile, and toxin content was determined using high-performance liquid chromatography-fluorescence detection (HPLC-FD).

[0099] Broilers in Group 2 were observed to have reduced diet and were depressed, and autopsy revealed enlarged livers. Broilers in Groups 1 and 3 showed no significant pathological features compared to those in Group 2.

[0100] Depend on Figure 4 It can be seen that compound plant essential oils can degrade or remove mycotoxins to a certain extent.

[0101] In summary, the composite plant essential oil prepared with cinnamon essential oil, peppermint essential oil, dandelion essential oil, garlic essential oil and citronella essential oil in a mass ratio of 1:(1-2):(2-4):(1-4):(2-4) can inhibit the growth of mold and alleviate the damage caused by mycotoxins to the body.

[0102] Example 4: Determination of the effect of different compound plant essential oils on the degradation of mycotoxins in vivo

[0103] 100 broiler chickens at 15 days old were fed the following diets and divided into 5 groups for 7 consecutive days.

[0104] Group 1: fed with a basic diet (AFB1≤50μg / kg, ZEA≤500μg / kg, DON≤5000μg / kg) and normal drinking water; Group 2: fed with a basic diet (moldy corn) and normal drinking water; Group 3: fed with a basic diet (moldy corn) and added with a composite plant essential oil (cinnamon essential oil, mint essential oil, dandelion essential oil) in an optimal ratio to drinking water; Group 4: fed with the same diet as Group 2 and added with a composite plant essential oil (cinnamon essential oil, mint essential oil, dandelion essential oil, garlic essential oil) in an optimal ratio to drinking water; Group 5: fed with the same diet as Group 2 and added with a composite plant essential oil (cinnamon essential oil, mint essential oil, dandelion essential oil, garlic essential oil, and citronella essential oil) in an optimal ratio to drinking water. On the eighth day, five chickens were taken from each group, dissected and the intestinal contents were collected. 1 g of sample was mixed with a methanol-water solution in a volume ratio of 85:15 to prepare a homogenate. The mixture was centrifuged and the upper methanol solution was taken. 10% sodium chloride was added and the mixture was extracted twice with chloroform. The chloroform layer was collected, dried under reduced pressure with nitrogen, dissolved in acetonitrile and the toxin content was determined using high performance liquid chromatography-fluorescence detector (HPLC-FD).

[0105] The broilers in Group 2 showed reduced diet and depression, and autopsy revealed enlarged livers. The broilers in the other groups showed no significant pathological changes compared to those in Group 2.

[0106] Depend on Figure 5 It can be seen that different composite plant essential oils can degrade or remove mycotoxins to a certain extent. With the increase in the amount of plant essential oil, the removal effect of mycotoxins improves, but there is no significant difference. This may be related to the synergistic effect of the plant essential oils. Therefore, in practical applications, different additions or ratios of composite plant essential oils can be selected based on the health status of livestock and poultry to reduce the damage caused by mold and its toxins to livestock and poultry.

[0107] Example 5: Determination of the effect of different compound plant essential oils on the degradation of aflatoxins B1, B2, G1, and G2 in vivo

[0108] 100 broiler chickens at 15 days old were randomly divided into 5 groups and fed for 7 consecutive days according to the following schedule:

[0109] Group 1: fed a basal diet with normal drinking water; Group 2: fed a basal diet (moldy corn) with normal drinking water; Group 3: fed a basal diet (moldy corn) with drinking water supplemented with an optimally formulated compound plant essential oil (cinnamon, peppermint, and dandelion); Group 4: fed the same diet as Group 2 with drinking water supplemented with an optimally formulated compound plant essential oil (cinnamon, peppermint, dandelion, and garlic); Group 5: fed the same diet as Group 2 with drinking water supplemented with an optimally formulated compound plant essential oil (cinnamon, peppermint, dandelion, garlic, and citronella). On day 8, five chickens from each group were killed and their intestinal contents collected for toxin content determination according to GB / T30955-2014.

[0110] Depend on Figure 6 It can be seen that different composite plant essential oils can degrade or remove aflatoxins to a certain extent. As the types of plant essential oils increase, the effect of removing mycotoxins gradually becomes stronger, and the inhibitory effect on AFB1 is the best. AFB1 is most widely and seriously contaminated in food crops (such as corn and peanuts) and processed products. Therefore, the present invention, through the compounding of multiple plant essential oils, targets food crops with the most serious AFB1 contamination, synergistically destroys the cell membrane structure of Aspergillus flavus, inhibits toxin synthesis (inhibition rate > 60%), and has no risk of chemical preservative residues.

[0111] Finally, it should be noted that the above 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 aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or make equivalent substitutions for some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A composite plant essential oil antibacterial composition, comprising an active ingredient and a pharmaceutically acceptable carrier, wherein the active ingredient comprises cinnamon essential oil, peppermint essential oil, and dandelion essential oil in a mass ratio of 1:1 to 2:2 to 4.

2. The composite plant essential oil antibacterial composition according to claim 1, wherein The active ingredients further include garlic essential oil and / or citronella essential oil; the mass ratio of the cinnamon essential oil, peppermint essential oil, dandelion essential oil, garlic essential oil and citronella essential oil is 1:1-2:2-4:1-4:2-4.

3. The composite plant essential oil antibacterial composition according to claim 1, wherein The pharmaceutically acceptable carrier includes an emulsifier, which is selected from at least one of Tween 80, propylene glycol, and tristyrylphenol polyoxyethylene ether, and the added amount of the emulsifier is 2 to 4 times the total weight of the active ingredient.

4. The composite plant essential oil antibacterial composition according to claim 2, wherein The content standards of the active ingredients in the composition are: cinnamaldehyde content ≥0.02%, menthol content ≥0.02%, taraxasterol content ≥0.02%, allicin content ≥0.06%, and geraniol content ≥0.03%.

5. A composite plant essential oil antibacterial agent, characterized in that: The invention contains the composite plant essential oil antibacterial composition according to any one of claims 1 to 4.

6. The composite plant essential oil antibacterial agent according to claim 5, characterized in that The antibacterial agent has an inhibitory effect on Aspergillus flavus and Fusarium graminearum.

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