Bactericidal composition, application and medicine for preventing and treating animal fungal dermatosis

Abstract

The invention provides a bactericidal composition, application and a medicine for preventing and treating animal fungal dermatosis, and relates to the technical field of animal fungal dermatosis. The main scheme of the invention is as follows: the bactericidal composition is prepared from the following components in percentage by mass: 10 to 30 weight percent of semen myristicae, 20 to 30 weight percent of root-bark of lindera angustifolia, 10 to 20 weight percent of herb of Indian adenosma, 5 to 10 weight percent of cortex pseudolaricis, 5 to 10 weight percent of rhizoma coptidis, 5 to 10 weight percent of radix scutellariae, 5 to 10 weight percent of cortex phellodendri, 5 to 10 weight percent of fructus cnidii and 5 to 10 weight percent of camellia bran. The invention also discloses application of the bactericidal composition as or in preparation of drugs for preventing and treating animal fungal dermatosis. The animal fungi comprise any one or more of epidermophyton, trichophyton, microsporum and malassezia. The bactericidal composition for preventing and treating animal fungal dermatosis has the advantages of long-acting effect, good stability and high safety.

Description

Technical Field

[0001] This invention relates to the field of animal fungal skin disease technology, and in particular to a bactericidal composition and its use, and a drug for preventing and treating animal fungal skin diseases. Background Technology

[0002] Fungal dermatitis in animals (such as pet fungal dermatitis) is a common disease in small animal clinics, mainly caused by pathogens such as Trichophyton, Epidermophyton, and Microsporum. It not only seriously affects the health of pets but can also be transmitted to humans through contact, posing a cross-species health threat. With the increasing number of pets, the incidence of these skin diseases continues to rise. Furthermore, the inappropriate use of antibiotics (such as ketoconazole and terbinafine) in clinical treatment has led to increasing drug resistance in pathogens, making prevention and treatment more challenging.

[0003] In existing technologies, products for preventing and treating fungal skin diseases in animals include chemically synthesized drugs, traditional Chinese medicine compositions, and bath and care products. For example, enconazole solution, as the first Class II new antifungal veterinary drug for pets in China, provides a new option for systemic treatment; pet bath products such as Pellafi Magic Wash and Chlorpyrifos have demonstrated their inhibitory effects on pathogenic fungi and bacteria of the skin through in vitro antibacterial tests. Traditional Chinese medicine compositions (such as compound preparations containing extracts of scorpion, turmeric, and kelp) or formulations containing camphor, dried ginger, and patchouli oil have also shown certain antifungal effects.

[0004] However, the existing products for the prevention and treatment of fungal skin diseases in animals still have obvious limitations: (1) Many topical preparations have a short duration of action and are difficult to maintain a long-term antibacterial effect on the skin surface; (2) Some products have poor component stability and are easily deactivated in high temperature environments; (3) Traditional drugs are difficult to balance efficient sterilization and safety, and long-term use may bring toxic side effects; (4) Few products can simultaneously target fungi remaining on the pet's body surface and in the environment for synergistic prevention and control.

[0005] Therefore, there is an urgent need to develop a bactericidal composition that has a long-lasting effect, good stability, high safety, and both therapeutic and preventive functions. This is of great significance for improving the clinical treatment effect of fungal skin diseases in animals, reducing the recurrence rate, and reducing the risk of zoonotic diseases. Summary of the Invention

[0006] In view of this, the present invention provides a bactericidal composition and its use, and a drug for preventing and treating fungal skin diseases in animals. The main objective is to provide a bactericidal composition for preventing and treating fungal skin diseases in animals that has a long-lasting effect, good stability, and high safety.

[0007] To achieve the above objectives, the present invention mainly provides the following technical solutions:

[0008] On one hand, embodiments of the present invention provide a bactericidal composition, wherein, by weight percentage, the bactericidal composition comprises: 10-30 wt% of *Eupatorium fortunei*, 20-30 wt% of *Gynostemma pentaphyllum*, 10-20 wt% of *Citrus aurantium*, 5-10 wt% of *Pseudolarix amabilis* bark, 5-10 wt% of *Coptis chinensis*, 5-10 wt% of *Scutellaria baicalensis*, 5-10 wt% of *Phellodendron chinense*, 5-10 wt% of *Cnidium monnieri*, and 5-10 wt% of *Camellia sinensis* bran.

[0009] This invention provides a bactericidal composition, wherein, by weight percentage, the bactericidal composition comprises: 20-50 wt% of *Eupatorium fortunei*, 10-20 wt% of *Houttuynia cordata*, 10-25 wt% of *Citrus aurantium*, 5-10 wt% of *Coptis chinensis*, 5-10 wt% of *Scutellaria baicalensis*, 5-10 wt% of *Phellodendron chinense*, 5-10 wt% of *Hibiscus mutabilis*, 5-10 wt% of *Valium valerian*, and 5-10 wt% of *Camellia sinensis* bran.

[0010] This invention provides a bactericidal composition, wherein, by weight percentage, the bactericidal composition comprises: 10-25 wt% of *Eupatorium fortunei*, 15-30 wt% of *Gynostemma pentaphyllum*, 10-25 wt% of *Citrus aurantium*, 5-10 wt% of *Coptis chinensis*, 5-10 wt% of *Scutellaria baicalensis*, 5-10 wt% of *Hibiscus mutabilis*, 5-10 wt% of *Valium valerian*, 5-10 wt% of *Cnidium monnieri*, and 10-20 wt% of *Camellia sinensis* bran.

[0011] On the other hand, the bactericidal composition described in any of the above claims is used as or in the preparation of a medicament for the prevention and treatment of fungal skin diseases in animals. Preferably, the animal fungi include any one or more of the genera *Epidermophytes*, *Trichophyton*, *Microsporum*, and *Malassezia*.

[0012] In another aspect, embodiments of the present invention provide a drug for preventing and treating fungal skin diseases in animals, wherein the drug for preventing and treating fungal skin diseases in animals is any of the bactericidal compositions described above; preferably, the animal fungi include any one or more of the genera *Epidermophyte*, *Trichophyton*, *Microsporum*, and *Malassezia*. Preferably, the bactericidal composition described above is applied to animals to prevent and treat fungal skin diseases in animals; preferably, the application method is any one of medicated bath, topical application, or smearing.

[0013] In another aspect, embodiments of the present invention provide a drug for preventing and treating fungal skin diseases in animals, wherein the drug for preventing and treating fungal skin diseases in animals comprises an extract of the bactericidal composition described in any one of the above-mentioned claims; preferably, the animal fungi include any one or more of the genera Epidermophyton, Trichophyton, Microsporum, and Malassezia.

[0014] Preferably, the extract is an extract solution; preferably, the extract solution comprises: 2-aminoeicosaeconane-1,5,7,19-tetraol, isobasanilide 4'-methyl ether-8-glucuronic acid, 6-hydroxykaempferol 7-glucuronide, glucosinolate-8-C-arabinopyranoside, 2,6-dimethoxy-1,4-benzoquinone, 3,4-dimethoxyphenylacetic acid, 3-O-methylquercetin, isobasanilide, and 4-aminosaeconyl benzoate. Rhamnin-7-O-glucoside, linalool, 5-aminolevulinic acid, 4',5,7-trihydroxy-3',3,6-trimethoxyflavone, 2-aminohexadecane-1,15-diol, 4-hydroxyphthalic acid, 2-aminohexadecane-1,4-diol, scutellarin, plantainin, ligustrazine lactone, 2-[3-(dihydroxyamino)-4-hydroxyphenyl]acetic acid, gardenia flavonoids, scutellarin.

[0015] Preferably, the method for preparing the extract includes the following steps: pulverizing the bactericidal composition, mixing it with water according to a set material-to-liquid ratio, soaking it at room temperature for a first set time, then ultrasonically treating it at a set temperature for a second set time, and filtering it to obtain the extract; preferably, the material-to-liquid ratio is 1:10-1:150; preferably, the first set time is 1-3 hours; preferably, the set temperature is 50-85℃; preferably, the second set time is 30-90 minutes.

[0016] Preferably, the extract is used to treat animals by bathing them to prevent or treat fungal skin diseases; or the extract is applied to the skin of animals to prevent or treat fungal skin diseases.

[0017] Compared with the prior art, the bactericidal composition and its use, and the drug for preventing and treating fungal skin diseases in animals of the present invention, have at least the following beneficial effects:

[0018] On one hand, embodiments of the present invention provide a bactericidal composition, which, by weight percentage, comprises: 10-30 wt% *Eupatorium fortunei*, 20-30 wt% *Gynostemma pentaphyllum*, 10-20 wt% *Citrus aurantium*, 5-10 wt% *Pseudolarix amabilis* bark, 5-10 wt% *Coptis chinensis*, 5-10 wt% *Scutellaria baicalensis*, 5-10 wt% *Phellodendron chinense*, 5-10 wt% *Cnidium monnieri*, and 5-10 wt% *Camellia sinensis* bran. On the other hand, embodiments of the present invention provide another bactericidal composition, which, by weight percentage, comprises: 20-50 wt% *Eupatorium fortunei*, 10-20 wt% *Gynostemma pentaphyllum*, 10-25 wt% *Citrus aurantium*, 5-10 wt% *Coptis chinensis*, 5-10 wt% *Scutellaria baicalensis*, 5-10 wt% *Phellodendron chinense*, 5-10 wt% *Hibiscus mutabilis*, 5-10 wt% *Valeriana officinalis*, and 5-10 wt% *Camellia sinensis* bran. This invention provides another bactericidal composition, which, by weight percentage, comprises: 10-25 wt% *Eupatorium fortunei*, 15-30 wt% *Gynostemma pentaphyllum*, 10-25 wt% *Citrus aurantium*, 5-10 wt% *Coptis chinensis*, 5-10 wt% *Scutellaria baicalensis*, 5-10 wt% *Hibiscus mutabilis*, 5-10 wt% *Valeriana officinalis*, 5-10 wt% *Cnidium monnieri*, and 10-20 wt% *Camellia sinensis* bran. Regarding the above three formulations of bactericidal compositions, the following explanation is provided: All three bactericidal compositions are derived from traditional Chinese herbal medicines and can significantly improve common skin symptoms in pets caused by fungi, such as tinea, eczema, dandruff, dermatitis, and mites. They are easy to use, have a pleasant aroma, and exhibit strong inhibitory effects on pathogenic fungi on the pet's body surface and in the surrounding environment. Cytotoxicity evaluation experiments have shown that the bactericidal compositions in this invention are relatively safe with few toxic side effects. Therefore, this invention provides a bactericidal composition with long-lasting effect, good stability, and high safety for the prevention and treatment of fungal skin diseases in animals.

[0019] On the other hand, embodiments of the present invention provide a drug for preventing and treating fungal skin diseases in animals, wherein the drug is a bactericidal composition as described in any of the above-mentioned embodiments; wherein the animal fungi include any one or more of the genera *Epidermophyte*, *Trichophyton*, *Microsporum*, and *Malassezia*. The bactericidal composition is used to treat animals by medicated bathing, thereby preventing and treating fungal skin diseases in animals. It should be noted that because this drug for preventing and treating fungal skin diseases in animals uses the aforementioned bactericidal composition, it has the advantages of long-lasting effect, good stability, high safety, and both therapeutic and preventative functions.

[0020] In another aspect, embodiments of the present invention provide a drug for preventing and treating fungal skin diseases in animals, wherein the drug comprises an extract of the bactericidal composition described in any of the above-mentioned embodiments; wherein the animal fungi include any one or more of the genera *Epidermophyte*, *Trichophyton*, *Microsporum*, and *Malassezia*. Preferably, the extract is an extract solution. The extract is used to treat animals by bathing them in a medicated bath; or the extract is applied to the skin of animals to prevent and treat fungal skin diseases. It should be noted that because this drug for preventing and treating fungal skin diseases in animals uses an extract of the above-mentioned bactericidal composition, it has the advantages of long-lasting effect, good stability, high safety, and both therapeutic and preventative functions.

[0021] The above description is merely an overview of the technical solution of the present invention. In order to better understand the technical means of the present invention and to implement it in accordance with the contents of the specification, the preferred embodiments of the present invention are described in detail below with reference to the accompanying drawings. Attached Figure Description

[0022] Figure 1 These are before-and-after comparison images of pets before and after medicated bathing with the bactericidal composition described in Example 1; wherein, Figure 1 The image numbered 1 is a photograph of the forelimb before the medicated bath; Figure 1 Photo number 2 in the image is a photograph of the forelimb 24 hours after the medicated bath. Figure 1 Photo number 3 is a photograph of the forelimb 72 hours after the medicated bath.

[0023] Figure 2 These are before-and-after comparison images of pets before and after medicated bathing with the bactericidal composition described in Example 2; wherein, Figure 2 The image numbered 1 is a photo of the upper part of the mouth before the medicated bath; Figure 2 Photo number 2 is a photo of the upper part of the mouth 24 hours after the medicated bath; Figure 2 Photo number 3 is a photo of the upper part of the mouth 72 hours after the medicated bath.

[0024] Figure 3 These are before-and-after comparison images of pets before and after medicated bathing with the bactericidal composition described in Example 3; wherein, Figure 3 The image numbered 1 is a photo of the back before the medicinal bath; Figure 3 Photo number 3 is a photo of the back taken 24 hours after the medicated bath; Figure 3 Photo number 3 is a photo of the back taken 72 hours after the medicated bath.

[0025] Figure 4 These are comparative images showing the pet before and after medicated bathing with the bactericidal composition described in Comparative Example 1; wherein, Figure 4The image numbered 1 is a photo of the back of the left foreleg before the medicinal bath; Figure 4 The image numbered 2 is a photo of the back of the left foreleg 24 hours after the medicated bath; Figure 4 The image numbered 3 is a photo of the back of the left foreleg 72 hours after the medicinal bath. Detailed Implementation

[0026] To further illustrate the technical means and effects adopted by the present invention to achieve the intended purpose, the specific embodiments, structures, features, and effects according to the present invention will be described in detail below with reference to the accompanying drawings and preferred embodiments. In the following description, different "embodiments" or "embodiments" do not necessarily refer to the same embodiment. Furthermore, specific features, structures, or characteristics in one or more embodiments can be combined in any suitable form.

[0027] During the research process, this invention proposed several bactericidal compositions, mainly as follows:

[0028] Formula 1: The bactericidal composition comprises, by weight percentage: 10-30 wt% of *Eupatorium fortunei*, 20-30 wt% of *Gynostemma pentaphyllum*, 10-20 wt% of *Citrus aurantium*, 5-10 wt% of *Pseudolarix amabilis* bark, 5-10 wt% of *Coptis chinensis*, 5-10 wt% of *Scutellaria baicalensis*, 5-10 wt% of *Phellodendron chinense*, 5-10 wt% of *Cnidium monnieri*, and 5-10 wt% of *Camellia sinensis* bran.

[0029] Formula 2: The bactericidal composition comprises, by weight percentage: 20-50 wt% of *Euphorbia hirta*, 10-20 wt% of *Houttuynia cordata*, 10-25 wt% of *Citrus aurantium*, 5-10 wt% of *Coptis chinensis*, 5-10 wt% of *Scutellaria baicalensis*, 5-10 wt% of *Phellodendron chinense*, 5-10 wt% of *Hibiscus mutabilis*, 5-10 wt% of *Vallisneria natans*, and 5-10 wt% of *Camellia sinensis* bran.

[0030] Formula 3: The bactericidal composition comprises, by weight percentage: 5-10 wt% of *Euphorbia hirta*, 5-15% of *Gynostemma pentaphyllum*, 5-15% of *Pseudolarix amabilis* bark, 5-15% of *Coptis chinensis*, 5-15% of *Hibiscus mutabilis* flower, 5-15% of *Vallisneria natans*, 5-15% of *Phellodendron chinense*, 5-15% of *Cnidium monnieri* fruit, and 5-15% of *Camellia sinensis* bran.

[0031] Formula 4: The bactericidal composition comprises, by weight percentage: 5-10% of *Euonymus fortunei*, 5-15% of *Citrus aurantium*, 5-12% of *Pseudolarix amabilis* bark, 5-12% of *Scutellaria baicalensis*, 5-12% of *Phellodendron chinense*, 5-12% of *Hibiscus mutabilis*, 5-12% of *Valium valerian*, 5-15% of *Cnidium monnieri*, and 5-20% of *Camellia sinensis* bran.

[0032] Formula 5: The bactericidal composition comprises, by weight percentage: 5-10% of *Eupatorium fortunei*, 5-15% of *Gynostemma pentaphyllum*, 5-12% of *Coptis chinensis*, 5-12% of *Scutellaria baicalensis*, 5-12% of *Phellodendron chinense*, 5-12% of *Hibiscus mutabilis*, 5-15% of *Valium valerian*, 5-12% of *Cnidium monnieri*, and 10-20% of *Camellia sinensis* bran.

[0033] Formula 6: The bactericidal composition comprises, by weight percentage: 1-5% of *Eupatorium fortunei*, 1-5% of *Lysimachia christinae*, 1-5% of *Cinnamomum camphora*, 5-15% of *Pseudolarix amabilis* bark, 5-15% of *Coptis chinensis*, 5-15% of *Scutellaria baicalensis*, 5-15% of *Hibiscus mutabilis*, 10-20% of *Vallisneria natans*, and 10-20% of *Camellia sinensis* bran.

[0034] Formula 7: The bactericidal composition comprises, by weight percentage: 5-10% of *Euphorbia hirta*, 5-15% of *Cinnamomum camphora*, 5-10% of *Pseudolarix amabilis* bark, 5-15% of *Coptis chinensis*, 5-15% of *Scutellaria baicalensis*, 5-15% of *Phellodendron chinense*, 5-15% of *Valeriana officinalis*, 5-15% of *Cnidium monnieri*, and 5-15% of *Camellia sinensis* bran.

[0035] Formula 8: The bactericidal composition comprises, by weight percentage: 10-25 wt% of *Eupatorium fortunei*, 15-30 wt% of *Gynostemma pentaphyllum*, 10-25 wt% of *Citrus aurantium*, 5-10 wt% of *Coptis chinensis*, 5-10 wt% of *Scutellaria baicalensis*, 5-10 wt% of *Hibiscus mutabilis*, 5-10 wt% of *Vallisneria natans*, 5-10 wt% of *Cnidium monnieri*, and 10-20 wt% of *Camellia sinensis* bran.

[0036] Among them, after extensive experimental verification, Formula 1, Formula 2, and Formula 8 showed the best efficacy in the prevention and treatment of fungal skin diseases in animals. All three formulas contain four components: antibacterial and antifungal, heat-clearing and dampness-removing, anti-inflammatory and antipruritic, and skin-moisturizing. Among them, *Eupatorium fortunei*, *Gynostemma pentaphyllum*, and *Citrus aurantium* are the main active ingredients. These three are rich in aromatic oils and flavonoids, possessing strong antibacterial activity and providing antibacterial, antipruritic, anti-inflammatory, and soothing effects when applied externally. *Pseudolarix amabilis* bark contains pseudolarix amabilis acid, tannins, and phenolic components, and has insecticidal, antifungal, and antipruritic effects. *Coptis chinensis*, *Scutellaria baicalensis*, and *Phellodendron chinense* are rich in alkaloids, and have heat-clearing and dampness-removing effects. *Cnidium monnieri* is rich in coumarins and volatile oils, and has antipruritic effects. *Hibiscus mutabilis* flower contains flavonoids, anthocyanins, and other components, and has anti-inflammatory, analgesic, pus-draining, and blood-cooling effects. *Valeriana officinalis* contains valeric acid, valerine, volatile oils, and other components, and has soothing, antibacterial, and blood-cooling effects. *Camellia sinensis* bran contains saponins, vitamin E, and minerals, and has the effects of improving dermatitis, decomposing dirt, and nourishing the skin and hair. The organic combination of nine kinds of Chinese herbal medicines achieves synergistic antibacterial, anti-inflammatory and antipruritic therapeutic effects.

[0037] The bactericidal compositions of Formulas 1, 2, and 8 can be used for direct medicated baths on animals to prevent and treat fungal skin diseases. Extracts (e.g., extracts) can also be extracted from the bactericidal compositions of Formulas 1, 2, and 8 for medicated baths on animals to prevent and treat fungal skin diseases; or the extracts can be applied to the skin of animals to prevent and treat fungal skin diseases.

[0038] Example 1

[0039] This embodiment provides a bactericidal composition, wherein, by mass percentage, the bactericidal composition comprises: 20% *Eupatorium fortunei*, 30% *Lysimachia christinae*, 15% *Citrus aurantium*, 5% *Pseudolarix amabilis* bark, 5% *Coptis chinensis*, 5% *Scutellaria baicalensis*, 5% *Phellodendron chinense*, 5% *Cnidium monnieri*, and 10% *Camellia sinensis* bran.

[0040] Example 2

[0041] This embodiment provides a bactericidal composition, wherein, by mass percentage, the bactericidal composition comprises: 25% *Eupatorium fortunei*, 25% *Gynostemma pentaphyllum*, 15% *Citrus aurantium*, 7% *Pseudolarix amabilis* bark, 6% *Coptis chinensis*, 5% *Scutellaria baicalensis*, 5% *Phellodendron chinense*, 5% *Cnidium monnieri*, and 7% *Camellia sinensis* bran.

[0042] Example 3

[0043] This embodiment provides a bactericidal composition, wherein, by mass percentage, the bactericidal composition comprises: 30% *Eupatorium fortunei*, 20% *Lysimachia christinae*, 15% *Citrus aurantium*, 9% *Pseudolarix amabilis* bark, 5% *Coptis chinensis*, 5% *Scutellaria baicalensis*, 5% *Phellodendron chinense*, 5% *Cnidium monnieri*, and 6% *Camellia sinensis* bran.

[0044] The formulations of Examples 1-3 satisfy Formulation 1.

[0045] Example 4

[0046] This embodiment provides a bactericidal composition, wherein, by mass percentage, the bactericidal composition comprises: 40% *Eupatorium fortunei*, 10% *Gynostemma pentaphyllum*, 20% *Citrus aurantium*, 5% *Coptis chinensis*, 5% *Scutellaria baicalensis*, 5% *Phellodendron chinense*, 5% *Hibiscus mutabilis*, 5% *Valium affine*, and 5% *Camellia sinensis* bran.

[0047] Example 5

[0048] This embodiment provides a bactericidal composition, wherein, by mass percentage, the bactericidal composition comprises: 35% *Eupatorium fortunei*, 15% *Gynostemma pentaphyllum*, 20% *Citrus aurantium*, 5% *Coptis chinensis*, 5% *Scutellaria baicalensis*, 5% *Phellodendron chinense*, 5% *Hibiscus mutabilis*, 5% *Valium affine*, and 5% *Camellia sinensis* bran.

[0049] Example 6

[0050] This embodiment provides a bactericidal composition, wherein, by mass percentage, the bactericidal composition comprises: 30% *Eupatorium fortunei*, 20% *Gynostemma pentaphyllum*, 20% *Citrus aurantium*, 5% *Coptis chinensis*, 5% *Scutellaria baicalensis*, 5% *Phellodendron chinense*, 5% *Hibiscus mutabilis*, 5% *Valium affine*, and 5% *Camellia sinensis* bran.

[0051] Among them, the formulations of Examples 4-6 satisfy Formulation 2.

[0052] Example 7

[0053] This embodiment provides a bactericidal composition, wherein, by mass percentage, the bactericidal composition comprises: 20% *Eupatorium fortunei*, 20% *Gynostemma pentaphyllum*, 15% *Citrus aurantium*, 6% *Coptis chinensis*, 6% *Scutellaria baicalensis*, 6% *Hibiscus mutabilis*, 6% *Valeriana officinalis*, 6% *Cnidium monnieri*, and 15% *Camellia sinensis* bran.

[0054] Example 8

[0055] This embodiment provides a bactericidal composition, wherein, by mass percentage, the bactericidal composition comprises: 15% *Eupatorium fortunei*, 25% *Gynostemma pentaphyllum*, 15% *Citrus aurantium*, 6% *Coptis chinensis*, 6% *Scutellaria baicalensis*, 6% *Hibiscus mutabilis*, 6% *Valeriana officinalis*, 6% *Cnidium monnieri*, and 15% *Camellia sinensis* bran.

[0056] Example 9

[0057] This embodiment provides a bactericidal composition, wherein, by mass percentage, the bactericidal composition comprises: 10% *Eupatorium fortunei*, 30% *Gynostemma pentaphyllum*, 15% *Citrus aurantium*, 6% *Coptis chinensis*, 6% *Scutellaria baicalensis*, 6% *Hibiscus mutabilis*, 6% *Valeriana officinalis*, 6% *Cnidium monnieri*, and 15% *Camellia sinensis* bran.

[0058] Among them, the formulations of Examples 7-9 satisfy Formulation 8.

[0059] Comparative Example 1

[0060] Comparative Example 1 provides a bactericidal composition, wherein, by mass percentage, the bactericidal composition comprises: 8% *Euphorbia hirta*, 12% *Gynostemma pentaphyllum*, 10% *Pseudolarix amabilis* bark, 12% *Coptis chinensis*, 12% *Hibiscus mutabilis* flower, 12% *Vallisneria natans*, 12% *Phellodendron chinense*, 7% *Cnidium monnieri*, and 15% *Camellia sinensis* bran.

[0061] Among them, the formula of Comparative Example 1 satisfies Formula 3.

[0062] Comparative Example 2

[0063] Comparative Example 2 provides a bactericidal composition, wherein, by mass percentage, the bactericidal composition comprises: 9% *Euonymus fortunei*, 12% *Citrus aurantium*, 10% *Pseudolarix amabilis* bark, 10% *Scutellaria baicalensis*, 10% *Phellodendron chinense*, 10% *Hibiscus mutabilis* flower, 10% *Valium affine*, 9% *Cnidium monnieri*, and 20% *Camellia sinensis* bran.

[0064] Among them, the formula of Comparative Example 2 satisfies Formula 4.

[0065] Comparative Example 3

[0066] Comparative Example 3 provides a bactericidal composition, wherein, by mass percentage, the bactericidal composition comprises: 10% of *Euphorbia hirta*, 10% of *Coptis chinensis*, 10% of *Scutellaria baicalensis*, 10% of *Phellodendron chinense*, 10% of *Hibiscus mutabilis*, 10% of *Valium affine*, 10% of *Cnidium monnieri*, and 20% of *Camellia sinensis* bran.

[0067] Among them, the formula of Comparative Example 3 satisfies Formula 5.

[0068] Comparative Example 4

[0069] Comparative Example 4 provides a bactericidal composition, wherein, by mass percentage, the bactericidal composition comprises: 5% *Eupatorium fortunei*, 5% *Lysimachia christinae*, 5% *Citrus aurantium*, 12% *Pseudolarix amabilis* bark, 10% *Coptis chinensis*, 10% *Scutellaria baicalensis*, 13% *Hibiscus mutabilis*, 20% *Vallisneria natans*, and 20% *Camellia sinensis* bran.

[0070] Among them, the formula of Comparative Example 4 satisfies Formula 6.

[0071] Comparative Example 5

[0072] Comparative Example 5 provides a bactericidal composition, wherein, by mass percentage, the bactericidal composition comprises: 9% *Eucommia ulmoides*, 11% *Citrus aurantium*, 8% *Pseudolarix amabilis* bark, 15% *Coptis chinensis*, 10% *Scutellaria baicalensis*, 10% *Phellodendron chinense*, 15% *Valeriana officinalis*, 10% *Cnidium monnieri*, and 12% *Camellia sinensis* bran.

[0073] Among them, the formula of Comparative Example 5 satisfies Formula 7.

[0074] The following experiments were conducted on the bactericidal compositions described in the above embodiments and comparative examples:

[0075] Experiment 1: Extraction of the bactericidal composition to obtain the extract.

[0076] There are four extraction methods:

[0077] Extraction Method 1: Immerse the sterilization composition in water (where the material-to-liquid ratio is 1:100), extract at room temperature for 3-48 hours, and filter to obtain the extract.

[0078] Extraction Method 2: Immerse the pulverized bactericidal composition in water (where the material-to-liquid ratio is 1:50), soak for 3-5 hours at room temperature, and then filter to obtain the extract.

[0079] Extraction method 3: Immerse the pulverized sterilization composition in water (wherein the material-to-liquid ratio is 1:25), soak at room temperature for 1-3 hours, then bathe in a water bath at 70-90℃ for 1-2 hours, and filter to obtain the extract.

[0080] Extraction method 4: Immerse the pulverized bactericidal composition in water (where the material-to-liquid ratio is 1:25), soak for 1-3 hours at room temperature, then sonicate at 70°C for 40 minutes, and filter to obtain the extract.

[0081] Thin-layer chromatography verified that extraction method 4 yielded substances with high abundance, making it the optimal extraction method.

[0082] Experiment 2: Metabolomics analysis of the extract

[0083] The bactericidal compositions of Examples 1-9 were extracted using extraction method 4 to obtain extracts; the metabolome of the extracts corresponding to the bactericidal compositions of Examples 1-9 was analyzed.

[0084] 1. The reagents and instruments used are shown in Tables 1 and 2.

[0085] Table 1: Information on Standards and Reagents

[0086]

[0087] Table 2: Instrument Information

[0088]

[0089] 2. Sample extraction process

[0090] (1) The biological sample (i.e., the extract sample) was placed in a freeze dryer and freeze-dried under vacuum for 63 h;

[0091] (2) Grind the powder using a grinder (30 Hz, 1.5 minutes);

[0092] (3) Weigh 30 mg of sample powder using an electronic balance, add 1500 μL of 70% methanol-water internal standard extraction solution pre-cooled at -20℃. The internal standard extraction solution is prepared by dissolving 1 mg of standard in 1 mL of 70% methanol-water to prepare a 1000 μg / mL standard stock solution, and further diluting the 1000 μg / mL stock solution with 70% methanol to prepare a 250 μg / mL internal standard solution.

[0093] (4) Vortex once every 30 minutes, each time lasting 30 seconds, for a total of 6 vortices;

[0094] (5) After centrifugation (12000 rpm, 3 minutes), aspirate the supernatant, filter the sample with a microporous membrane, and store it in a sample vial for UPLC-MS / MS analysis.

[0095] 3. Chromatography-mass spectrometry acquisition conditions

[0096] The data acquisition instrument system mainly includes ultra-high performance liquid chromatography and tandem mass spectrometry.

[0097] Liquid phase conditions mainly include:

[0098] (1) Column: Agilent SB-C18 1.8µm, 2.1 mm * 100mm;

[0099] (2) Mobile phase: Phase A is ultrapure water (with 0.1% formic acid added), and Phase B is acetonitrile (with 0.1% formic acid added).

[0100] (3) Elution gradient: The proportion of phase B is 5% at 0.00 min, and increases linearly to 95% within 9.00 min and is maintained at 95% for 1 min. From 10.00 to 11.10 min, the proportion of phase B decreases to 5% and is balanced at 5% until 14 min.

[0101] (4) Flow rate 0.35 mL / min; column temperature 40℃; injection volume 2 μL.

[0102] Mass spectrometry conditions mainly include:

[0103] The electrospray ionization source temperature was 500°C; the ion spray voltage (IS) was 5500 V (positive ion mode) / -4500 V (negative ion mode); ion source gas I, gas II, and curtain gas were set to 50, 60, and 25 psi, respectively, and the collision-induced ionization parameter was set to high. QQQ scans were performed using MRM mode with the collision gas (nitrogen) set to medium. DP and CE for each MRM ion pair were completed through further optimization of declustering voltage and collision energy. A specific set of MRM ion pairs was monitored at each epoch based on the metabolites eluted within each epoch.

[0104] 4. Test Results

[0105] A total of 5213 metabolites were detected in the extract of the composition, belonging to 13 classes of compounds including flavonoids, phenolic acids, lipids, alkaloids, terpenes, amino acids, quinones, lignans, coumarins, nucleotides, organic acids, tannins, and steroids. Broad-target metabolomics analysis of the extract revealed the top 20 compounds with the highest relative abundance, as shown in Table 3.

[0106] Table 3

[0107]

[0108] Experiment 3: Antifungal effect of bactericidal composition extract on skin fungi

[0109] Extraction methods 4 were used to extract the bactericidal compositions of Examples 1-9 and Comparative Examples 1-5 to obtain extracts. Antifungal tests were then conducted on the extracts corresponding to Examples 1-9 and Comparative Examples 1-5.

[0110] The experimental steps are as follows: Take a 96-well culture plate, dilute the sample to be tested (extract sample), add fungal culture to each well, and the final concentration is 5 × 10⁻⁶. 5 CFU / mL, cultured at 25 °C for 5 days, and absorbance was measured at 625 nm using a microplate reader. A blank control of the culture medium, a fungal control, and a positive drug control were also included in the experiment.

[0111] The results of the antifungal test of the extract are shown in Table 4:

[0112] Table 4

[0113]

[0114] Table 4 shows that Examples 1-9 showed strong inhibitory effects (inhibition rate greater than 80%), while the comparative examples showed relatively strong inhibitory effects (inhibition rate greater than 50%).

[0115] Experiment 4: Cytotoxicity Evaluation Experiment of the Composition

[0116] Extraction method 4 was used to extract the bactericidal compositions of Examples 1, 4, and 7 to obtain extracts. Cytotoxicity evaluation experiments were then performed on the extracts corresponding to those of Examples 1, 4, and 7.

[0117] 1. Cells

[0118] Adult dermal fibroblasts (HDFa) were purchased from Beina Biotechnology; mouse melanoma cells (B16) were purchased from the Cell Resource Center of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences.

[0119] 2. Reagents

[0120] DMEM (high glucose) medium, PBS, Hank's balanced salt solution (HBSS), penicillin, streptomycin, and fetal bovine serum (FBS) were purchased from Hyclone; 0.25% trypsin (containing EDTA) was purchased from Gibico; and MTS reagent was purchased from Promega.

[0121] 3. Experimental Methods

[0122] HDFa cells were mixed with the sample solution (extraction solution) in a 96-well cell culture plate and serially diluted. Three replicate wells were set up, and a blank control without the drug was also set up. The cells were cultured at 37°C and 5% CO2 for 24 h. Cytotoxicity was detected by MTS colorimetric assay, and the OD value was measured by a microplate reader at a wavelength of 490 nm. Cell viability was calculated.

[0123] Cell viability (%) = OD490nm of experimental wells / OD490nm of blank wells × 100%

[0124] CC50 (50% cytotoxic concentration) is calculated using the Reed & Muench method.

[0125] 4. The experimental results are as follows:

[0126] The toxic effects of the bactericidal composition on adult dermal fibroblasts (HDFa) are shown in Table 5.

[0127] Table 5

[0128]

[0129] The toxic effects of the bactericidal composition on mouse melanoma cells (B16) are shown in Table 6.

[0130] Table 6

[0131]

[0132] Cytotoxicity evaluation experiments revealed that the bactericidal composition of this invention is relatively safe and has few toxic side effects.

[0133] Experiment 5: Results of the application of bactericidal composition extract in the treatment of pet skin diseases

[0134] Extraction methods 4 were used to extract the bactericidal compositions of Example 1, Example 4, Example 7, and Comparative Example 1 to obtain extracts.

[0135] The pets were given a medicated bath using extracts from the bactericidal compositions in Examples 1, 4, 7, and Comparative Example 1. The specific operation consisted of the following four steps:

[0136] The first step, soothing and grooming: Before bathing, gently comb your pet's fur, untangle any knots, and gently soothe it.

[0137] The second step is a local test: Apply a small amount of the diluted medication to a small area of ​​thin skin, such as behind the pet's ear or the inner thigh, using a cotton swab. Wait 15-20 minutes and observe for any redness, swelling, or allergic reactions. If no abnormalities are observed, a full-body medicated bath can then be administered.

[0138] Step 3, Bathing: 1. Medicated Bath: Gently place your pet in the diluted medicated solution. Starting from the back, use a spoon or cup to scoop up the solution and slowly pour it over your pet, ensuring that all fur and skin are thoroughly soaked. Gently massage the skin, especially the affected areas, to promote absorption. Absolutely avoid getting the solution in the eyes, ears, mouth, and nose. You can wipe the head with a damp towel when washing. 2. Soaking: Let your pet soak in the medicated solution for 5-10 minutes. During this time, continue to soothe your pet; you can give it treats to distract it. 3. Drying / Rinsing: You can skip rinsing with water and simply dry your pet with a towel to retain the medicated solution's effectiveness. If you are concerned about your pet licking the solution, rinse it thoroughly with warm water. 4. Thoroughly Drying and Blow-drying: Use a large towel to absorb as much moisture as possible. Then, use a hairdryer on a low heat or cool setting to thoroughly dry the fur until the skin is completely dry as well.

[0139] Step 4, Post-bath observation: Closely observe your pet's skin condition and mental state for the next 24-72 hours to see if the symptoms improve or if any adverse reactions occur.

[0140] The experimental results are as follows:

[0141] Figure 1 These are before-and-after comparison images of pets before and after medicated bathing with the bactericidal composition described in Example 1; wherein, Figure 1 The image numbered 1 is a photo of the forelimbs of a Corgi before a medicated bath. The affected dog has hair loss and bald patches on its paws and frequently licks and scratches the affected area. It is caused by Microsporum. Figure 1 Photo number 2 is a photo of the forelimbs 24 hours after the medicated bath; the dog's licking and scratching of the affected area decreased. Figure 1 Photo number 3 is a photo of the forelimb 72 hours after the medicated bath. The area of ​​bald patches on the affected skin has shrunk, and the dog's licking and scratching of the affected area has significantly decreased, indicating a marked improvement in symptoms.

[0142] Figure 2 These are before-and-after comparison images of pets before and after medicated bathing with the bactericidal composition described in Example 4; wherein, Figure 2 The picture number 1 is a photo of the upper part of the mouth of a Bichon Frise before the medicated bath. The affected dog has erythematous hair loss patches on its mouth and frequently scratches the affected area, which is caused by Trichophyton mentagrophytes. Figure 2 Photo number 2 is a photo of the upper part of the mouth of a Bichon Frise 24 hours after the medicated bath. The dog's scratching behavior on the affected area has decreased. Figure 2 Photo number 3 shows the upper part of the Bichon Frise's mouth 72 hours after the medicated bath. The area of ​​hair loss on the affected skin has shrunk, the dog's scratching behavior has significantly decreased, and the symptoms have clearly improved.

[0143] Figure 3These are before-and-after comparison images of pets before and after medicated bathing with the bactericidal composition described in Example 7; wherein, Figure 3 The image numbered 1 is a photograph of the body of a French Bulldog before a medicated bath. The dog's skin is covered with dandruff and it is scratching itself, which is caused by the overgrowth of Malassezia. Figure 3 Photo number 2 is a photo of the back of a French Bulldog 24 hours after a medicated bath. The dandruff has been significantly reduced and the coat is shiny. Figure 3 Photo number 3 in the image is a photograph of the body of a French Bulldog 72 hours after the medicated bath, showing significant improvement in symptoms.

[0144] Figure 4 These are comparative images showing the pet before and after medicated bathing with the bactericidal composition described in Comparative Example 1; wherein, Figure 4 The image numbered 1 is a photo of the back of the forelimb of a Corgi before a medicated bath. The affected dog has hair loss, papules and scabs, indicating a cross-infection of Microsporum canis, Microsporum gypseum, and bacteria. Figure 4 The picture number 2 in the picture is a photo of the back of the forelimb of the Corgi 24 hours after the medicated bath. The papules have improved and the skin surface has become smooth. Figure 4 The picture number 3 is a photo of the back of the forelimb of a Corgi 72 hours after the medicated bath. The skin on the affected area has scabbed over, but the symptoms persist and the effect is not obvious.

[0145] In summary, the bactericidal compositions provided in the embodiments of the present invention are all derived from commonly used folk herbs, which can significantly improve common pet skin symptoms such as tinea, eczema, dandruff, dermatitis and mites caused by fungi. They are easy to use, have a pleasant fragrance, and have a strong inhibitory effect on pathogenic fungi on the pet's body surface and in the surrounding environment. Cytotoxicity evaluation experiments have shown that the bactericidal compositions of the embodiments of the present invention are relatively safe and have few toxic side effects.

[0146] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Any simple modifications, equivalent changes, and alterations made to the above embodiments based on the technical essence of the present invention shall still fall within the scope of the technical solution of the present invention.

Claims

1. A fungicidal composition, characterized by, The fungicidal composition comprises, in percentage by mass: Yuzixiang 10-30 wt%, Wanglangcao 20-30 wt%, Dadaochen 10-20 wt%, Tujiangpi 5-10 wt%, Huanglian 5-10 wt%, Huangqin 5-10 wt%, Huangbai 5-10 wt%, Shechuangzi 5-10 wt%, Shanchaigu 5-10 wt%.

2. A fungicidal composition, characterized by, The fungicidal composition comprises, in percentage by mass: Yuzixiang 20-50 wt%, Wanglangcao 10-20 wt%, Dadaochen 10-25 wt%, Huanglian 5-10 wt%, Huangqin 5-10 wt%, Huangbai 5-10 wt%, Furonghua 5-10 wt%, Weixiao 5-10 wt%, Shanchaigu 5-10 wt%.

3. A fungicidal composition, characterized by, The fungicidal composition comprises, in percentage by mass: Yuzixiang 10-25 wt%, Wanglangcao 15-30 wt%, Dadaochen 10-25 wt%, Huanglian 5-10 wt%, Huangqin 5-10 wt%, Furonghua 5-10 wt%, Weixiao 5-10 wt%, Shechuangzi 5-10 wt%, Shanchaigu 10-20 wt%.

4. Use of the fungicidal composition of any one of claims 1-3 as or in the preparation of a medicament for the prevention and treatment of fungal skin diseases in animals; preferably, the fungal skin diseases in animals include any one or several of Epidermophyton, Trichophyton, Microsporum, Malassezia.

5. A medicine for preventing and treating fungal dermatosis of an animal, characterized by, The medicament for the prevention and treatment of fungal skin diseases in animals is the fungicidal composition of any one of claims 1-3; preferably, the fungal skin diseases in animals include any one or several of Epidermophyton, Trichophyton, Microsporum, Malassezia.

6. The medicine for preventing and treating fungal dermatosis of animals according to claim 5, wherein The fungicidal composition of any one of claims 1-3 is applied to animals for the prevention and treatment of fungal skin diseases in animals; preferably, the application is any one of bath, external application, and smearing.

7. A medicament for preventing and treating fungal dermatosis of an animal, characterized by, The medicament for the prevention and treatment of fungal skin diseases in animals comprises an extract of the fungicidal composition of any one of claims 1-3; preferably, the fungal skin diseases in animals include any one or several of Epidermophyton, Trichophyton, Microsporum, Malassezia.

8. The medicine for preventing and treating fungal dermatosis of animals according to claim 7, characterized in that, The extract is an extract solution; Preferably, the extract solution comprises 2-aminoeicosan-1,5,7,19-tetrol, isohyperoside 4'-methyl ether-8-glucuronide, 6-hydroxykaempferol 7-glucuronide, glucoside-8-C-arabinopyranoside, 2,6-dimethoxy-1,4-benzoquinone, 3,4-dimethoxyphenylacetic acid, 3-O-methylquercetin, isorhamnetin-7-O-glucoside, casuarinin, 5-amino-ketopentanoic acid, 4',5,7-trihydroxy-3',3,6-trimethoxyflavone, 2-aminohexadecan-1,15-diol, 4-hydroxyphthalic acid, 2-aminohexadecan-1,4-diol, inuloxin, plantaginin, senkyunolide, 2-[3-(dihydroxyamino)-4-hydroxyphenyl]acetic acid, geniposide, and baicalin.

9. The medicine for preventing and treating fungal dermatosis of animals according to claim 8, wherein The preparation method of the extract solution comprises the following steps: The bactericidal composition is crushed, mixed with water according to a set material-to-liquid ratio, soaked at room temperature for a first set time, ultrasonically treated at a set temperature for a second set time, filtered, and an extract is obtained; Preferably, the material-to-liquid ratio is 1:10-1:150; Preferably, the first set time is 1-3 hours; Preferably, the set temperature is 50-85°C; Preferably, the second set time is 30-90 minutes.

10. The medicine for preventing and treating fungal dermatosis of animals according to claims 7-9, characterized in that, The extract is used for medicated bath of animals to prevent and treat fungal skin diseases of animals; or The extract is applied to the skin of animals to prevent and treat fungal skin diseases of animals.

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