Antibacterial agent containing vitex extract and use thereof

Abstract

The present application relates to the technical field of plant extract application, and particularly relates to a bacteriostatic agent containing vitex extract and application thereof.The main active ingredient of the bacteriostatic agent is vitex extract and zanthoxylum bungeanum extract.It is found for the first time that the composition of vitex extract and zanthoxylum bungeanum extract can effectively inhibit Acidovorax citrulli and Acidovorax avenae subsp.avenae.

Description

Technical Field

[0001] This invention generally relates to the field of plant extract application technology, and more specifically, to an antibacterial agent containing Vitex negundo extract and its application. Background Technology

[0002] Vitex negundo L., also known as thorn bush, is a plant belonging to the genus Vitex in the family Verbenaceae. It is widely distributed in the southern provinces of China. Vitex negundo is a shrub or small tree with quadrangular twigs covered in grayish-white down. It has five palmate compound leaves, rarely three; the leaves are oblong-lanceolate to lanceolate, with an acuminate apex, green on the upper surface and densely covered with grayish-white down on the lower surface. The flowers are terminal, arranged in cymose panicles; the calyx is bell-shaped, and the corolla is pale purple with fine hairs. The drupe is nearly spherical, about 2 mm in diameter. Flowering occurs from April to June, and fruiting from July to October. Vitex negundo fruit, the dried, mature fruit of Vitex negundo, is a widely used traditional Chinese medicine. It has a pungent and bitter taste and was originally used for its antipyretic, expectorant, digestive, and gas-relieving effects.

[0003] Previous studies have shown that extracts of Vitex negundo have inhibitory effects on Gram-positive bacteria, Gram-negative bacteria, yeast, and Penicillium, with significant inhibitory effects on Staphylococcus aureus and Moraxella catarrhalis. The dichloromethane extract of Vitex negundo seeds also exhibits insecticidal activity against various pests. However, existing research on the effective antibacterial chemical components and their activities in Vitex negundo is scarce, leading to a lack of clarity regarding its antibacterial mechanism. Consequently, research on the antibacterial effects of its extracts is incomplete. These factors make it particularly difficult to explore the combined use of Vitex negundo extracts with other antibacterial components, resulting in a lack of relevant research and documentation. Summary of the Invention

[0004] The first aspect of the present invention relates to an antibacterial agent, the main active ingredients of which are Vitex negundo extract and Zanthoxylum bungeanum extract.

[0005] Optionally, the Vitex negundo extract is an aqueous extract of Vitex negundo callus tissue.

[0006] Optionally, the Sichuan pepper extract is an alcoholic extract of Sichuan pepper.

[0007] Optionally, the mass concentration ratio of the Vitex negundo extract to the Zanthoxylum bungeanum extract is 1:(2.5-7.5).

[0008] Optionally, the main active ingredient of the antibacterial agent also includes scutellarin.

[0009] Optionally, based on the purity of the scutellaria baicalensis flavonoids being 80%, the mass concentration ratio of the Vitex negundo extract, Zanthoxylum bungeanum extract, and scutellaria baicalensis flavonoids is Vitex negundo extract: Zanthoxylum bungeanum extract: scutellaria baicalensis flavonoids = 1:(2.5-7.5):(18-22).

[0010] Optionally, the antibacterial agent is formulated as a suspension, emulsion, microemulsion, emulsifiable concentrate, wettable powder, or water-dispersible granule.

[0011] A second aspect of the invention relates to an agricultural composition comprising the antibacterial agent as described above, further comprising additional components in addition to the main active ingredient of the antibacterial agent, said additional components being agriculturally effective in an amount selected from compounds or compositions of: nutrients, fertilizers, acaricides, bactericides, fungicides, insecticides, microbial agents, nematicides, and insecticides.

[0012] A third aspect of the invention relates to the use of the antibacterial agents described above, or the agricultural compositions described above, in the prevention and control of diseases in plants caused by infection with Acidovorax citrulli and / or Acidovorax avenae subsp. avenae.

[0013] Optionally, the method of applying the antibacterial agent or the agricultural composition includes: applying it topically or to the whole plant to be treated, or coating it on plant seeds, or spreading it through fertilizer, or spreading it through irrigation, or a combination of the above methods.

[0014] The fourth aspect of the invention relates to the use of the antibacterial agents described above, or the agricultural compositions described above, in the control of pests caused by plant-derived tea geometrid moths, cabbage caterpillars, or aphids.

[0015] A fifth aspect of the present invention relates to an animal insecticide comprising the aforementioned antibacterial agent, further comprising additional components other than the main active ingredient of the antibacterial agent, said additional components being an insecticidal or acaricidal effective amount selected from the group consisting of: fungicides, acaricides, tapeworms, and nematicides.

[0016] The sixth aspect of the invention relates to the use of the above-described antibacterial agent for animal use in the treatment of fungal diseases, mite diseases, scabies, nematode diseases and / or tapeworm diseases in animals.

[0017] Optionally, the animal being treated may be a cow, goat, sheep, cat, dog, or rabbit.

[0018] This invention is the first to discover that the combination of Vitex negundo extract and Zanthoxylum bungeanum extract can effectively inhibit Acidovorax citrulli and Acidovorax avenae subsp. avenae; and in a more preferred embodiment, the combination of the two with Scutellaria baicalensis flavonoids will have a significant synergistic effect on the prevention and control of Acidovorax avenae subsp. avenae. Detailed Implementation

[0019] Reference will now be made to detailed embodiments of the present invention, one or more of which are described below. Each example is provided for explanation and not for limitation of the invention. In fact, it will be apparent to those skilled in the art that various modifications and variations can be made to the invention without departing from its scope or spirit. For example, features described or illustrated as part of one embodiment may be used in another embodiment to produce further embodiments.

[0020] Unless otherwise stated, all terms used to disclose this invention (including technical and scientific terms) have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Further guidance is provided below for a better understanding of the teachings of this invention. The terminology used herein in the specification of this invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

[0021] The terms "and / or," "or / and," and "and / or" as used herein include any one of two or more of the related listed items, as well as any and all combinations of the related listed items. These arbitrary and all combinations include any two related listed items, any more related listed items, or a combination of all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or," "or / and," and "and / or," it should be understood that in this application, the technical solution undoubtedly includes technical solutions connected by "logical AND," and also undoubtedly includes technical solutions connected by "logical OR." For example, "A and / or B" includes three parallel solutions: A, B, and A+B. For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, a technical solution that is connected by "logical OR"), as well as any and all combinations of A, B, C, and D, that is, combinations of any two or three of A, B, C, and D, and also combinations of all four of A, B, C, and D (that is, a technical solution that is connected by "logical AND").

[0022] The terms “containing,” “comprising,” and “including” as used in this invention are synonyms and are inclusive or open-ended, not excluding additional, uncited members, elements, or method steps.

[0023] In this invention, the numerical range represented by endpoints includes all numerical values ​​and fractions contained within that range, as well as the endpoints mentioned.

[0024] This invention relates to concentration values, which include fluctuations within a certain range. For example, fluctuations are allowed within a corresponding precision range. For instance, 2% can fluctuate within ±0.1%. For larger values ​​or values ​​that do not require overly precise control, even greater fluctuations are permitted.

[0025] In this invention, "(preparation)" and "composition" are equivalent and refer to a composition of substances suitable for agricultural or horticultural use.

[0026] In this invention, the terms "multiple" or "various" are used unless otherwise specified, referring to a quantity of 2 or more.

[0027] In this invention, the technical features described in an open-ended manner include both closed-ended technical solutions composed of the listed features and open-ended technical solutions that include the listed features.

[0028] In this invention, terms such as "preferred," "better," "more suitable," and "ideal" merely describe implementation methods or embodiments with better effects and should be understood not to limit the scope of protection of this invention. In this invention, terms such as "optionally," "optionally," and "optional" mean that something is optional, that is, selected from either "with" or "without" a parallel solution. If multiple "optional" statements appear in a technical solution, unless otherwise specified and without contradiction or mutual constraint, each "optional" statement is independent.

[0029] The first aspect of the present invention relates to an antibacterial agent, the main active ingredients of which are Vitex negundo extract and Zanthoxylum bungeanum extract.

[0030] In some embodiments, the Vitex negundo extract is an aqueous extract of Vitex negundo callus tissue.

[0031] In some embodiments, the Sichuan pepper extract is an alcoholic extract of Sichuan pepper. For this alcoholic extract, a suitable alcohol concentration can be 70%–95%. A typical extraction method is, for example, to crush the Sichuan pepper, soak it in a mixture of raw material and 70%–95% alcohol at a mass-to-volume ratio of (0.5–1.5) g : (0.5–1.5) mL, for 4–8 hours (preferably 6 hours), and then separate the solid and liquid phases to obtain the supernatant.

[0032] In some embodiments, the mass concentration ratio of the Vitex negundo extract to the Zanthoxylum bungeanum extract is 1:(2.5-7.5). Preferred mass concentration ratios may also be 1:3, 1:3.5, 1:4, 1:4.5, 1:5, 1:5.5, 1:6, or 1:6.5.

[0033] In some embodiments, the main active ingredient of the antibacterial agent also includes scutellarin.

[0034] In some embodiments, based on the purity of the scutellaria baicalensis flavonoids being 80%, the mass concentration ratio of the Vitex negundo extract, Zanthoxylum bungeanum extract, and scutellaria baicalensis flavonoids is Vitex negundo extract: Zanthoxylum bungeanum extract: scutellaria baicalensis flavonoids = 1:(2.5-7.5):(18-22). Preferred mass concentration ratios may also be 1:(2.5-5):(18-22), 1:(2.5-7.5):(19-21), 1:(2.5-5):(19-21), 1:(2.5-7.5):20, or 1:(2.5-5):20.

[0035] The above antibacterial agents can be formulated into suspensions, emulsions, microemulsions, emulsifiable concentrates, wettable powders, or water-dispersible granules using conventional methods in this field, depending on actual needs.

[0036] Furthermore, according to the general selection of those skilled in the art, the antimicrobial agents of the present invention may optionally contain one or more agents, such as, but not limited to, emulsifiers, wetting agents, tension modifiers, preservatives, buffers, and antioxidants. Tension modifiers used in the antimicrobial agents of the present invention include, but are not limited to, salts such as sodium acetate, sodium chloride, potassium chloride, mannitol, or glycerol, and other pharmaceutically acceptable tension modifiers. Preservatives used in the antimicrobial agents described herein include, but are not limited to, benzalkonium chloride, chlorobutanol, thimerosal, phenylmercuric acetate, and phenylmercuric nitrate. Various buffers and methods for adjusting the pH that can be used to prepare the antimicrobial agents include, but are not limited to, acetate buffers, citrate buffers, phosphate buffers, and borate buffers. Similarly, antioxidants used in the antimicrobial agents are well known in the art and include, for example, sodium metabisulfite, sodium thiosulfate, acetylcysteine, butylated hydroxyanisole, and butylated hydroxytoluene. Flavonoids are compounds found in plants and are known to have a variety of beneficial biochemical and antioxidant activities.

[0037] Another aspect of the present invention relates to an agricultural composition comprising the antibacterial agent as described above, further comprising additional components in addition to the main active ingredient of the antibacterial agent, said additional components being agriculturally effective in an amount selected from compounds or compositions of: nutrients, fertilizers, acaricides, bactericides, fungicides, insecticides, microbial agents, nematicides, and insecticides.

[0038] The agricultural compositions referred to in this invention also include those for horticultural use.

[0039] Surfactants, binders, stabilizers, etc., commonly used as adjuvants in agricultural compositions, can be used alone or in combination as needed. Stabilizers such as antioxidants and / or pH adjusters can be used. Light stabilizers may also be used in certain situations.

[0040] The total content of these additives can be from 0 wt% to 80 wt%, and the content of the carrier is the value obtained by subtracting the content of the active ingredient and additives from 100 wt%.

[0041] In some specific embodiments, the binder is selected from one or more of the following substances: hydroxypropyl methylcellulose, ethylcellulose, methylcellulose, carboxymethylcellulose, polyvinyl alcohol, polyvinylpyrrolidone, polyoxyethylene and its copolymers, latex, polyamide, sugar, glucose, maltose, starch, lignin sulfonate, guar gum, urea, alginate, polysaccharide, aqueous polyester, polyether, epoxy resin, isocyanate, ethylene-vinyl acetate copolymer, polyacrylate latex and its copolymer latex. The amount of binder added is industrially acceptable, preferably 1% to 20%, 1.5% to 15%, 2% to 7%, or 5% of the composition by mass.

[0042] In some specific embodiments, the additives include one or more of the following: sodium dodecylbenzenesulfonate, sodium butylnaphthalenesulfonate, trehalose, glycerin, sodium ligninsulfonate, sodium alkylnaphthalenesulfonate condensate, nicotinic acid, alcohol, buffer salt, sodium chloride, amino acids, vitamins, proteins, polypeptides, polysaccharides or monosaccharides, yeast extract, precipitated silica, tea saponin, and skim milk.

[0043] The method for preparing the agricultural composition as described above involves mixing the components together, and optionally includes compaction, drying, and granulation.

[0044] In some embodiments, the particle size of the agricultural composition may be similar to that of commercially available dry fertilizer products, for example, 0.1 cm to 3 cm, or 0.3 cm to 2 cm, or 0.5 cm to 1 cm.

[0045] Another aspect of the present invention relates to the use of the antibacterial agents described above, or the agricultural compositions described above, in the prevention and control of diseases in plants caused by infection with Acidovorax citrulli and / or Acidovorax avenae subsp. avenae.

[0046] In some embodiments, the method of applying the antibacterial agent or the agricultural composition includes: applying it topically or to the whole plant to be treated, or coating it on plant seeds, or spreading it through fertilizer, or spreading it through irrigation, or a combination of the above methods.

[0047] The plants involved in this invention are typically natural hosts of watermelon acidophilus and / or cucurbit fruit spot pathogens, and may include watermelon (Citrullus lanatus), cantaloupe (Cucumis melo), thick-skinned cantaloupe (Cucuumis melo var. cantalupensis, including Hami melon and Gashi melon), squash (Cucurbita pepo), cucumber (Cucumissativus L.), zucchini (CuCurbita pepo), honeydew melon (Cucuumis melo var. inodorus), and bitter melon (Momordica charantia), etc.

[0048] The embodiments of the present invention will be described in detail below with reference to examples. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. For experimental methods in the following embodiments where specific conditions are not specified, please refer to the guidelines given in this invention, or follow experimental manuals or conventional conditions in the art, or other experimental methods known in the art, or follow the conditions recommended by the manufacturer.

[0049] In the specific embodiments described below, the measurement parameters involving raw material components may have slight deviations within the weighing accuracy range unless otherwise specified. Temperature and time parameters are subject to acceptable deviations due to instrument testing accuracy or operational precision.

[0050] Example 1: Obtaining Vitex negundo callus and preparing its extract

[0051] 1. Culture of Vitex negundo callus

[0052] The Vitex negundo callus tissue used in this invention can be obtained by known methods or any existing technology capable of obtaining Vitex negundo callus tissue, such as the methods described in Chinese patent documents with application numbers 202011604196.7 or 202011604180.6.

[0053] In this embodiment, the callus tissue was prepared by the following method.

[0054] 1) Materials

[0055] On a clear summer morning, collect semi-lignified paper strips with 4-6 axillary buds from healthy, mature Vitex negundo plants as explants.

[0056] After rinsing the explants, disinfect their surface. Soak in 70% alcohol solution for 30 seconds, 0.1% HgCl for 25 minutes, rinse 5 times with sterile water, and then cut into stem segments of 2-5 cm in length with 2-3 axillary buds.

[0057] 2) Culture medium and culture conditions

[0058] The experiment used 1 / 2 MS as the basal medium, with a sucrose concentration of 30 g / L, an agar concentration of 10 g / L, and a pH of 6.1. 2,4-D 1.2 μM, 6-BA 0.20 μM, TDZ 0.20 μM, vitamin C 25 mg / L, and inositol 30 mg / L were added. The culture temperature was (25±2)℃, the light intensity was 2100 lx, the photoperiod was 12 h, and the relative humidity was 55%. Callus tissue was harvested on day 10 of culture.

[0059] 2. Preparation of Vitex negundo extract

[0060] Take 2.0-2.5 g of fresh Vitex negundo callus tissue obtained after the above culture, weigh accurately, grind and crush it using a grinder (60 Hz, 60 s × 2 times), then add 10 times the volume of deionized water, extract by ultrasonication at 700 W for 20 min, centrifuge at 5000 rpm for 30 min, collect the supernatant, filter at 0.45 μM to obtain Vitex negundo extract. Based on 20 extractions, the average solid content of the extract is 4.36 ± 0.78 mg / mL.

[0061] Example 2: Inhibitory effects of different plant extracts on cucurbit pathogens

[0062] 1. Experimental Materials

[0063] The cucurbit pathogens used in this embodiment were cultured using the methods shown in the table below.

[0064]

[0065] The plant extracts used in this embodiment were obtained from the following sources:

[0066] Vitex negundo extract (prepared in Example 1, diluted to a concentration of 1 g / mL);

[0067] Ginger extract (weigh 5.00g of ginger powder, use 100mL of 80% ethanol as the extraction solution, reflux in a Soxhlet extractor until the liquid in the extractor is colorless, filter, and obtain a yellow extract. Perform parallel extractions, combine the extracts, evaporate to dryness using a rotary evaporator, weigh, and then use 50% ethanol as the solvent to prepare a ginger extract solution with a mass concentration of 1g / mL; the ginger powder was purchased from a supermarket).

[0068] Garlic oil (100% purity, purchased from Shanghai Shuangyuan Garlic Oil Co., Ltd.; prepared as a solution with a mass concentration of 1 g / mL);

[0069] Scutellaria baicalensis flavonoids (80% purity, provided by Shaanxi Fusen Biotechnology Co., Ltd., prepared as a solution with a mass concentration of 1g / mL);

[0070] Thyme essential oil (100% purity, purchased from Shanghai Huien International Trade Co., Ltd., prepared as a solution with a mass concentration of 1g / mL);

[0071] Soapberry extract (wash soapberries with water, crush them into a paste, soak them for 6 hours at a ratio of 100g raw material: 100mL 95% alcohol, centrifuge, and concentrate the supernatant to a mass concentration of 1g / mL);

[0072] Sichuan pepper extract (wash Sichuan pepper clean with water, crush it into a paste, soak it for 6 hours at a ratio of 100g raw material: 100mL 95% alcohol, centrifuge, and concentrate the supernatant to a mass concentration of 1g / mL).

[0073] Berberine (purity 97%, purchased from Shaanxi Senfu Biotechnology Co., Ltd., prepared as a solution with a mass concentration of 1 g / mL);

[0074] Paeoniflorin extract (red peony root is crushed into coarse powder, and 4-20 times the weight of red peony root is added as the extraction solution for each extraction. It is extracted by hot reflux 6 times, soaking for 1 hour each time. The extract is centrifuged and purified by passing it through a macroporous adsorption resin column and a polyamide column in sequence. Impurities are first washed off with water, and then eluted with 10%-90% ethanol. The ethanol eluent is collected, concentrated, and then loaded onto a polyamide column for elution with water. The eluent is concentrated to a mass concentration of 1 g / mL).

[0075] Each of the above extracts was transferred into a 10mL stoppered graduated test tube, sealed, and stored in a refrigerator (0-4℃) for later use.

[0076] 2. Experimental Methods

[0077] 1) Determination of the inhibitory effects of single-component plant extracts on watermelon acidophilus and cucurbit fruit spot pathogens.

[0078] To measure the inhibitory effects of various plant extracts on watermelon acidophilus and cucurbit fruit spot pathogens, each plant extract was set up as a group with 8 replicates per group. The concentration of the plant extracts was determined based on their solubility and the antibacterial ability at different concentrations, with tetracycline as a positive control.

[0079] The antibacterial activity was assessed using the Oxford cup method. The prepared bacterial suspension was diluted to 10⁻⁶ with sterile physiological saline. 6CFU / mL, take 200mL and spread it evenly on the surface of MH agar medium. Then, place Oxford cups vertically on the medium surface (4 Oxford cups on each 9cm diameter petri dish), and gently press them down to ensure they are in contact with the medium without gaps. Add 100mL of plant extract solution to 3 of the cups to form 3 parallels, and add physiological saline to the other cup as a blank control. Then, incubate at 37℃ for 24 hours, and measure the diameter of the inhibition zone with calipers.

[0080] First, a single-factor, completely randomized experimental design was employed. Based on the antibacterial effects of the single plant extracts mentioned above, plant extracts with good inhibitory effects against both *Acidic bacteria* of watermelon and *Fungiella fuciformis*, a pathogen causing fruit spot disease in cucurbits, were selected.

[0081] The test results for the two pathogens are shown in Tables 1 and 2.

[0082] Table 1. Diameter of the inhibition zone (mm) of plant extracts against *Acidobacterium moniliforme*.

[0083]

[0084] Note: No inhibition zones were observed in garlic oil, thyme essential oil, and paeoniflorin extract.

[0085] Table 2. Diameter of the inhibition zone (mm) of plant extracts against cucurbit fruit spot pathogens.

[0086]

[0087] Note: No inhibition zones were observed in ginger extract, garlic oil, thyme essential oil, saponin extract, berberine, and paeoniflorin extract.

[0088] Combining the results in Tables 1 and 2, only Vitex negundo extract, Zanthoxylum bungeanum extract, and Scutellaria baicalensis flavonoids showed significant antibacterial effects against both pathogens, with Vitex negundo extract and Zanthoxylum bungeanum extract showing significantly better effects. Both bacteria tested are common pathogens of cucurbits and often cause co-infections. Therefore, considering ease of application, Vitex negundo extract, Zanthoxylum bungeanum extract, and Scutellaria baicalensis flavonoids were selected for a compound experiment. Based on the experimental results and adhering to the principle of minimizing concentration while maximizing antibacterial effect, the initial concentration of Scutellaria baicalensis flavonoids was set at 100 mg / mL, the initial concentration of Zanthoxylum bungeanum extract was set at 25 mg / mL, and the initial concentration of Vitex negundo extract was set at 10 mg / mL.

[0089] The specific compounding experiments were arranged as follows: 100 mg / mL and 200 mg / mL of Scutellaria baicalensis flavonoids; 25 mg / mL, 50 mg / mL, and 75 mg / mL of Zanthoxylum bungeanum extract; and 10 mg / mL of Vitex negundo extract served as the control group. The compounding ratios of Vitex negundo extract and Scutellaria baicalensis flavonoids were 1:10 (concentration ratio 10:100) and 1:20 (concentration ratio 10:200); the compounding ratios of Vitex negundo extract and Zanthoxylum bungeanum extract were 1:2.5 (concentration ratio 10:25), 1:5 (concentration ratio 10:50), and 1:7.5 (concentration ratio 10:75); and the compounding ratio of Vitex negundo extract, Scutellaria baicalensis flavonoids, and Zanthoxylum bungeanum extract was 1:2.5. Seventeen groups were used, with six replicates per group. The experimental results are shown in Table 3. The ratios were: 1:10 (concentration ratio 10:25:100), 1:2.5:20 (concentration ratio 10:25:200), 1:5:10 (concentration ratio 10:50:100), 1:5:20 (concentration ratio 10:50:200), 1:7.5:10 (concentration ratio 10:75:100), and 1:7.5:20 (concentration ratio 10:75:200).

[0090] Table 3. Inhibitory effects of plant extract combination on *Acidic bacteria of watermelon* and *Fungiella fuciformis*, the pathogen causing fruit spot disease in cucurbits.

[0091]

[0092]

[0093] Note: Different lowercase letters in the superscript of data in the same column indicate significant differences (P<0.05).

[0094] For watermelon acid bacteria, a compound ratio of 1:2.5 of Vitex negundo extract and Zanthoxylum bungeanum extract can achieve the ideal antibacterial effect, and the two show a relatively obvious synergistic effect. However, further increasing the amount of Scutellaria baicalensis flavonoids does not effectively increase its antibacterial effect.

[0095] Surprisingly, a highly synergistic effect was achieved when combining extracts of Vitex negundo, Zanthoxylum bungeanum, and Scutellaria baicalensis flavonoids to treat fruit spot disease in cucurbits. The optimal ratio of 1:2.5:20 yielded the best cost-effective inhibitory effect. This combination effectively inhibited the proliferation of two common pathogenic microorganisms affecting cucurbits and fruits. Furthermore, as a natural extract, it is environmentally friendly and shows great promise for application.

[0096] Example 3: Agricultural Composition

[0097] Raw materials: Vitex negundo extract (same as in Example 2), Zanthoxylum bungeanum extract (same as in Example 2), Scutellaria baicalensis flavonoids (same as in Example 2);

[0098] Bacillus thuringiensis pesticide (active ingredient concentration 2000 IU / ul, purchased from Shandong Tainuo Pharmaceutical Co., Ltd.);

[0099] Preparation method: Take 100ml of Bacillus thuringiensis pesticide, and fully dissolve Vitex negundo extract, Zanthoxylum bungeanum extract, Scutellaria baicalensis flavonoids and Bacillus thuringiensis pesticide in solvent at a mass concentration ratio of 1:2.5:18:50. Add emulsifier, dispersant and suspending agent and stir thoroughly. Then slowly add antifreeze dissolved in water and stir at high speed to obtain water emulsion, which is the agricultural composition of this experimental group.

[0100] Example 4: Agricultural Composition

[0101] Raw materials: Vitex negundo extract (same as in Example 2), Zanthoxylum bungeanum extract (same as in Example 2), Scutellaria baicalensis flavonoids (same as in Example 2);

[0102] Indoxacarb pesticide (purchased from DuPont, USA)

[0103] Preparation method: Take 100ml of indoxacarb, and fully dissolve Vitex negundo extract, Zanthoxylum bungeanum extract, Scutellaria baicalensis flavonoids and Bacillus thuringiensis pesticide in solvent at a mass concentration of 1:7.5:22:80. Add emulsifier, dispersant and suspending agent and stir thoroughly. Then slowly add antifreeze dissolved in water and stir at high speed to obtain water emulsion, which is the agricultural composition of this embodiment.

[0104] Example 5: Application of agricultural composition in the control of plant-derived pests

[0105] 1. Test insects: Larvae of tea geometrid moth, cabbage caterpillar, and aphid were collected from the field and reared indoors using fresh tea leaves that had not been exposed to pesticides. They were fed fresh fruits and vegetables, and the water source was tap water. The temperature was 25℃±1℃. The relative humidity was 75%±5%, and the photoperiod L:D = 10:12h. The fourth instar larvae were used for the experiment.

[0106] 2. Experimental group:

[0107] Experimental Group 1: Agricultural Composition of Example 3

[0108] Experimental Group 2: Agricultural Composition of Example 4

[0109] Blank control group 1: 100 ml of Bacillus thuringiensis pesticide was taken;

[0110] Blank control group 2: Take 100ml of indoxacarb.

[0111] 3. Test methods:

[0112] Place 2-3 tea leaves in culture bottles, and inoculate each bottle with 15 fourth-instar larvae of tea geometrid moth, cabbage caterpillar, and aphid. Incubate under constant temperature and light (25℃±1℃, photoperiod L:D=10:12h). Dilute experimental groups 1 and 2 with water to 100mg / L, and apply 100ul of each larva to the head and neck of the larvae in the culture bottle using a drip method. Observe and record the mortality rate of the test insects every 2 days. The mortality rate is calculated as follows: Mortality rate (%) = (Number of live insects before treatment - Number of live insects after treatment) / Number of live insects before treatment × 100. Blank control group 1 and blank control group 2 are respectively dripped with the same dose of Bacillus thuringiensis pesticide and indoxacarb.

[0113] 4. Test Results and Analysis:

[0114] The toxicity and lethality of the agricultural composition of this embodiment to larvae at different concentrations are shown in Table 4.

[0115] Table 4

[0116]

[0117] As shown in Table 4, the agricultural composition of the present invention has a high mortality rate against tea geometrid moth, cabbage caterpillar, and aphid. The mortality rate can reach more than 95% after 5 days of use. Compared with traditional Bacillus thuringiensis pesticides and indoxacarb, the mortality rate is significantly improved. It can be seen that Vitex negundo extract, Zanthoxylum bungeanum extract, and Scutellaria baicalensis flavonoids have an insecticidal synergistic effect on plant-derived pests, especially showing significant control effects against tea geometrid moth, cabbage caterpillar, and aphid.

[0118] Example 6: Animal Insecticide

[0119] Raw materials: Vitex negundo extract (same as in Example 2), Zanthoxylum bungeanum extract (same as in Example 2), Scutellaria baicalensis flavonoids (same as in Example 2);

[0120] Praziquantel (purchased from Suzhou Medison Pharmaceutical Co., Ltd.);

[0121] Penetration enhancers: Lauric acid, dimethyl sulfoxide

[0122] Solvents: Glycerin, Polyethylene Glycol

[0123] Antioxidants: Ascorbate palmitate, butylated hydroxytoluene

[0124] Preparation method: Take 10g of praziquantel, dissolve Vitex negundo extract, Zanthoxylum bungeanum extract, Scutellaria baicalensis flavonoids and praziquantel in glycerol and polyethylene glycol in a mass ratio of 1:4:18:10, add lauric acid, dimethyl sulfoxide, ascorbate palmitate and dibutylhydroxytoluene and stir at high speed until uniform, thus obtaining the animal insecticide of this embodiment.

[0125] Example 7: Animal Insecticide

[0126] Raw materials: Vitex negundo extract (same as in Example 2), Zanthoxylum bungeanum extract (same as in Example 2), Scutellaria baicalensis flavonoids (same as in Example 2);

[0127] Avermectin (purchased from Zhejiang Jiuzhou Pharmaceutical Co., Ltd.);

[0128] Penetration enhancers: linoleic acid, lauric acid

[0129] Solvents: Glycerin, Propylene Glycol

[0130] Antioxidant: Ascorbate palmitate

[0131] Preparation method: Take 10g of abamectin, dissolve Vitex negundo extract, Zanthoxylum bungeanum extract, Scutellaria baicalensis flavonoids and Bacillus thuringiensis pesticide in glycerol and propylene glycol in a mass ratio of 1:5:22:6, then add linoleic acid, lauric acid and ascorbyl palmitate and stir at high speed to obtain the animal insecticide of this embodiment.

[0132] Example 8: Application of animal insecticides in the treatment of fungal diseases, mite infestations, scabies, nematode infestations, or tapeworm infestations in animals.

[0133] 1. Test animals: Golden Retriever (5.5 months old)

[0134] 2. Therapeutic agents: The animal insecticide of Example 6:

[0135] 3. Symptoms before treatment: Itchy skin, frequent scratching of the tail, legs and other parts of the body, eczema, mild redness and ulceration in some areas, and severe hair loss on the soles of the feet, tail and other parts of the body;

[0136] 4. Treatment and Efficacy: The animal insecticide from Example 6 was diluted 50 times and used to thoroughly wash the Golden Retriever, soaking it for 15-20 minutes. After drying, the fur was blow-dried. This was done once every 3 days. After one week of continuous use, the above symptoms completely disappeared. The Golden Retriever's living environment was then thoroughly disinfected by spraying. The symptoms were completely controlled with no recurrence. Using the same dose of praziquantel as a blank control, the dog was washed and soaked for 15-20 minutes, then the fur was blow-dried. This was done once every 3 days. After two weeks of continuous use, symptoms of itching and hair loss persisted. After four weeks of continuous use, the symptoms largely disappeared, but recurrence was observed.

[0137] 5. Conclusion and Analysis: Experiments have shown that the animal insecticide of the present invention has a special therapeutic effect on fungal skin diseases, hair loss, mites, scabies, eczema, nematode disease or tapeworm disease in dogs, and has a rapid onset of action and a cure rate of over 99%.

[0138] Example 9: Application of animal insecticides in the treatment of fungal diseases, mite infestations, scabies, nematode infestations, or tapeworm infestations in animals.

[0139] 1. Test animals: dogs and cats (3.5 months old)

[0140] 2. Therapeutic agents: The animal insecticide of Example 7:

[0141] 3. Symptoms before treatment: scratching, hair loss, oval-shaped insects visible in the hair, localized tinea, and small red pustules visible on the abdomen;

[0142] 4. Treatment and Efficacy: Dilute the animal insecticide in Example 7 30 times, then thoroughly wash and soak the dog or cat for 10-20 minutes. After drying, use a hairdryer to dry the fur. Use once every two days. After six consecutive days of use, the above symptoms completely disappeared. Thoroughly disinfect the dog or cat's living environment by spraying. The symptoms were completely controlled with no recurrence. Using the same dose of avermectin as a blank control, after washing and soaking for 10-20 minutes and drying the fur, use once every two days. After 1-2 weeks of continuous use, symptoms such as hair loss and local red spots and pustules still appeared.

[0143] 5. Conclusion and Analysis: Experiments have shown that the animal insecticide of the present invention has a special therapeutic effect on fungal skin diseases, hair loss, mites, scabies, eczema, nematode disease or tapeworm disease in dogs, and has a rapid onset of action and a cure rate of over 99%.

[0144] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims, and the specification can be used to interpret the content of the claims.

Claims

1. An antibacterial agent, characterized in that, The active ingredients are Vitex negundo extract, Zanthoxylum bungeanum extract, and Scutellaria baicalensis flavonoids; Based on the purity of 80% for the scutellaria baicalensis flavonoids, the mass concentration ratio of the Vitex negundo extract, Zanthoxylum bungeanum extract, and scutellaria baicalensis flavonoids is Vitex negundo extract: Zanthoxylum bungeanum extract: scutellaria baicalensis flavonoids = 1:(2.5~7.5):(18~22). The Vitex negundo extract is an aqueous extract of Vitex negundo callus tissue; The Sichuan pepper extract is an alcoholic extract of Sichuan pepper; the alcohol is 70%~95% alcohol. The scutellaria baicalensis flavonoids were obtained from Shaanxi Fusen Biotechnology Co., Ltd.

2. The antibacterial agent according to claim 1, characterized in that, The dosage forms are suspensions, emulsions, microemulsions, emulsifiable concentrates, wettable powders, or water-dispersible granules.

3. An agricultural composition comprising the antibacterial agent of claim 1 or 2, characterized in that, It also contains additional components other than the antibacterial active ingredient, said additional components being agriculturally effective amounts of compounds or combinations selected from: fertilizers, bactericides, fungicides, and insecticides.

4. The antibacterial agent according to claim 1 or 2, or the agricultural composition according to claim 3, for controlling plant damage caused by *Acidobacterium hygrophilum*. Acidovorax citrulli and / or cucurbit fruit spot pathogens Acidovorax avenae subsp. avenae Applications in diseases caused by infection.

5. The application according to claim 4, characterized in that, The method of applying the antibacterial agent or the agricultural composition includes: applying the agent locally or to the whole plant to be treated, or spreading it through fertilizer, or spreading it through irrigation, or a combination of the above methods.

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