A bacterial agent with deodorizing effect, and a preparation method and application thereof

Abstract

The present application relates to a kind of bacterial agent with deodorization effect and its preparation method and application, the strain in the bacterial agent includes the Bacillus megaterium YBG02 strain with the preservation number CCTCC NO:M2021052 and the Bacillus subtilis YBS01 strain with the preservation number CCTCC NO:M2021053.The Bacillus megaterium and Bacillus subtilis used in the present application are all safe strains to human, animal, environment, have obvious removal effect on odor, and the bacterial agent prepared can be widely applied in cat litter production, with good temperature resistance and low loss rate.

Description

Technical Field

[0001] This invention belongs to the field of microbial technology and relates to a deodorizing bacterial agent, its preparation method, and its application. Background Technology

[0002] While dogs and cats relieve stress and meet the emotional needs of their owners, they also create some cleaning challenges for the home environment. According to the 2022 China Pet Industry White Paper, odor is the biggest core pain point for pet-owning families, accounting for 59.5%. Typical odors include ammonia (NH3), hydrogen sulfide (H2S), and indole skatole, which smell like feces. These odorous substances are volatile and have a low odor threshold. Prolonged exposure to odorous environments can easily harm the human eyelids, nasal and oral cavities, respiratory mucosa, digestive system, and cardiovascular system.

[0003] Currently, cat litter deodorizers, both domestically and internationally, are mainly classified into four categories: physical, chemical, plant-based, and biological. Physical deodorizers primarily use activated carbon and mineral salts, adsorbing odors through a porous structure. They require regular replacement, are less efficient, but offer higher safety. Chemical deodorizers are the most widely used on the market, boasting advantages such as fast deodorization, strong adsorption capacity, and good results, but they can be irritating and have lower safety. Plant-based deodorizers use plant-derived extracts, such as essential oils and wood vinegar, primarily achieving deodorization through absorption and masking. They offer good safety but can be somewhat irritating, and their long-lasting effect is less than ideal. Biological deodorizers utilize the addition of functional microorganisms, whose metabolism effectively degrades common odor factors, addressing the root cause of odors and reducing their production in odor-generating sources. They are safe, environmentally friendly, non-toxic, long-lasting, and non-irritating, becoming active upon contact with the odor source and remaining dormant after the source is removed. In recent years, with the advancement of biotechnology and the continuous improvement of the fermentation industry chain, biological deodorizers have emerged as a rising star, experiencing explosive growth. Their green, safe, and pollution-free characteristics have made them a new choice in the deodorization field. Therefore, providing a biological agent with deodorizing effects has become an urgent problem to be solved in this field. Summary of the Invention

[0004] In view of the shortcomings of the prior art, the purpose of this invention is to provide a microbial agent with deodorizing effect, its preparation method and application.

[0005] To achieve this objective, the present invention employs the following technical solution:

[0006] In a first aspect, the present invention provides a microbial agent with deodorizing effect, wherein the strains in the microbial agent include Bacillus megaterium YBG02 strain with accession number CCTCC NO: M2021052 and Bacillus subtilis YBS01 strain with accession number CCTCC NO: M2021053.

[0007] The deodorizing agent prepared by this invention has high stability, low loss rate, and good deodorizing effect. The strains used are all Bacillus species that are safe for humans, animals, and the environment and have wide applications. Bacillus megaterium YBG02 and Bacillus subtilis YBS01 have a certain synergistic effect in improving the deodorizing effect. The odors that can be removed include ammonia, hydrogen sulfide, or indole.

[0008] Preferably, the ratio of viable Bacillus megaterium YBG02 strain to Bacillus subtilis YBS01 strain is (1-2):(1-2).

[0009] The specific point values ​​in (1-2) can all be selected from 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, etc. Other specific point values ​​within the above range can also be selected, which will not be elaborated here.

[0010] Preferably, the viable count of the bacterial agent is not less than 2 × 10⁻⁶. 10 CFU / mL or 2×10 10 CFU / g, for example 2×10 10 CFU / mL, 3×10 10 CFU / mL, 4×10 10 CFU / mL, 5×10 10 CFU / mL, 6×10 10 CFU / mL, 7×10 10 CFU / mL, 8×10 10 CFU / mL, 9×10 10 CFU / mL, 1×10 11 CFU / mL, 1×10 12 CFU / mL or 2×10 10 CFU / g, 3×10 10 CFU / g, 4×10 10 CFU / g, 5×10 10 CFU / g, 6×10 10 CFU / g, 7×10 10CFU / g, 8×10 10 CFU / g, 9×10 10 CFU / g, 1×10 11 CFU / g, 1×10 12 CFU / g, etc., other specific point values ​​within the above range can be selected, and will not be elaborated on here.

[0011] Preferably, the bacterial strains in the bacterial agent also include Lactiplantibacillus Plantarum ATCC8014 and Saccharomyces cerevisiae ATCC9763.

[0012] The addition of Lactobacillus plantarum and Saccharomyces cerevisiae can further enhance the deodorizing effect of the microbial agent, and Saccharomyces cerevisiae and Lactobacillus plantarum have potential synergistic effects in the above-mentioned aspects.

[0013] Preferably, the ratio of viable counts of the Bacillus megaterium YBG02 strain, Bacillus subtilis YBS01 strain, Lactiplantibacillus Plantarum ATCC8014 strain, and Saccharomyces cerevisiae ATCC9763 strain is (10-20):(10-20):1:1.

[0014] The specific point values ​​in (10-20) can be selected from 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, etc. Other specific point values ​​within the above range can also be selected, which will not be elaborated here.

[0015] In a second aspect, the present invention provides a method for preparing a microbial agent with deodorizing effect according to the first aspect, the preparation method comprising: activating the bacterial strain, fermenting it and mixing it with a protective agent, freeze-drying it, and mixing the resulting microbial powder with a carrier to obtain the product.

[0016] Preferably, the carrier comprises talc powder.

[0017] Thirdly, the present invention provides the application of the deodorizing microbial agent according to the first aspect in the preparation of deodorizing cat litter.

[0018] Preferably, the cat litter includes tofu cat litter, cassava cat litter, wood chip cat litter, tea leaf cat litter, or coconut coir cat litter.

[0019] Fourthly, the present invention provides a cat litter with deodorizing effect, wherein the raw materials for preparing the cat litter include pea residue, corn starch, guar gum and the deodorizing microbial agent described in the first aspect.

[0020] Preferably, the cat litter comprises, by weight, 30-50 parts pea residue, 20-50 parts corn starch, and 1-5 parts guar gum, and by live bacteria, 1×10⁻⁶ of the deodorizing microbial agent described in the first aspect. 7 CFU / g-2×10 7 CFU / g.

[0021] The mass fractions of pea pulp can be selected from 30, 32, 35, 38, 40, 42, 45, 48, and 50 parts, etc. The mass fractions of corn starch can be selected from 20, 22, 25, 28, 30, 32, 35, 38, 40, 42, 45, 48, and 50 parts, etc. The mass fractions of guar gum can be selected from 1, 2, 3, 4, and 5 parts, etc. The viable count of the microbial agent can be selected from 1×10⁻⁶. 7 CFU / g, 1.1×10 7 CFU / g, 1.2×10 7 CFU / g, 1.3×10 7 CFU / g, 1.4×10 7 CFU / g, 1.5×10 7 CFU / g, 1.6×10 7 CFU / g, 1.7×10 7 CFU / g, 1.8×10 7 CFU / g, 1.9×10 7 CFU / g, 2×10 7 CFU / g, etc., other specific point values ​​within the above range can be selected, and will not be elaborated on here.

[0022] Fifthly, the present invention provides a method for preparing cat litter with deodorizing effect according to the fourth aspect, the method comprising: physically mixing the raw materials, granulating, and drying to obtain the litter.

[0023] Preferably, the granulation temperature is 75-85℃, such as 75℃, 77℃, 80℃, 82℃, 85℃, etc. Other specific values ​​within the above range can be selected, and will not be elaborated here.

[0024] Preferably, the moisture content of the raw materials is 38-42%, such as 38%, 39%, 40%, 41%, 42%, etc. Other specific values ​​within the above range can be selected, and will not be elaborated here.

[0025] Preferably, the moisture content of the cat litter is below 12%, such as 1%, 2%, 4%, 6%, 8%, 10%, 12%, etc. Other specific values ​​within the above range can be selected, and will not be elaborated here.

[0026] Compared with the prior art, the present invention has the following beneficial effects:

[0027] The deodorizing agent prepared by this invention has high stability, low loss rate, and good deodorizing effect. The strains used are all Bacillus species that are safe for humans, animals, and the environment and have wide applications. Bacillus megaterium YBG02 and Bacillus subtilis YBS01 have a certain synergistic effect in improving the deodorizing effect. The odors that can be removed include ammonia, hydrogen sulfide, or indole. Detailed Implementation

[0028] To further illustrate the technical means and effects of the present invention, the following describes the technical solution of the present invention in conjunction with preferred embodiments of the present invention. However, the present invention is not limited to the scope of the embodiments.

[0029] The sources of the active ingredients in the products involved in the following examples and comparative examples are as follows (only the active ingredients are shown; other necessary excipients contained in commercially available raw materials are not described):

[0030] The Bacillus megaterium strain YBG02 mentioned below is classified as Bacillus megaterium YBG02, with accession number CCTCC NO: M2021052, accession date of January 13, 2021, deposited at the China Center for Type Culture Collection (CCTCC) at Wuhan University, Wuhan, China, and produced by Shandong Kangdian Biotechnology Co., Ltd.

[0031] The following information pertains to the Bacillus subtilis strain YBS01. Classified as Bacillus subtilis YBS01, its accession number is CCTCC NO: M2021053, the accession date is January 13, 2021, the depositary institution is the China Center for Type Culture Collection, the depositary address is Wuhan University, Wuhan, China, and the production company is Shandong Kangdian Biotechnology Co., Ltd.

[0032] Pea pulp, corn starch, and guar gum are produced by Jiangsu Zhenghao Pet Products Co., Ltd.

[0033] Example 1

[0034] This embodiment provides a microbial agent with deodorizing effect, wherein the microbial agent comprises Bacillus megaterium strain YBG02 and Bacillus subtilis strain YBS01 with a live bacteria ratio of 1:1, and the total live bacteria count is 2×10⁻⁶. 10 CFU / g.

[0035] The preparation method is as follows: after activating the bacterial strain, fermenting, centrifuging, and freeze-drying, the effective viable count of the bacterial powder is 3×10⁻⁶. 11 CFU / g, obtained by mixing 1 part bacterial powder with 14 parts talc.

[0036] Example 2

[0037] This embodiment provides a microbial agent with deodorizing effect, comprising Bacillus megaterium strain YBG02 and Bacillus subtilis strain YBS01 with a live bacteria ratio of 2:1, and a total live bacteria count of 2 × 10⁻⁶. 10 CFU / g.

[0038] The preparation method is the same as in Example 1.

[0039] Example 3

[0040] This embodiment provides a microbial agent with deodorizing effect, wherein the microbial agent comprises Bacillus megaterium strain YBG02 and Bacillus subtilis strain YBS01 with a live bacteria ratio of 1:2, and the total live bacteria count is 2×10⁻⁶. 10 CFU / g.

[0041] The preparation method is the same as in Example 1.

[0042] Example 4

[0043] This embodiment provides a microbial agent with deodorizing effect. The microbial agent includes Bacillus megaterium strain YBG02, Bacillus subtilis strain YBS01, Lactobacillus plantarum strain ATCC8014, and Saccharomyces cerevisiae strain ATCC9763 with a live bacteria count ratio of 10:10:1:1, and the total live bacteria count is 2×10⁻⁶. 10 CFU / g.

[0044] The preparation method is the same as in Example 1.

[0045] Example 5

[0046] This embodiment provides a microbial agent with deodorizing effect. The microbial agent includes Bacillus megaterium strain YBG02, Bacillus subtilis strain YBS01, Lactobacillus plantarum strain ATCC8014, and Saccharomyces cerevisiae strain ATCC9763 with a live bacteria count ratio of 20:10:1:1, and the total live bacteria count is 2×10⁻⁶. 10 CFU / g.

[0047] The preparation method is the same as in Example 1.

[0048] Example 6

[0049] This embodiment provides a microbial agent with deodorizing effect. The microbial agent includes Bacillus megaterium strain YBG02, Bacillus subtilis strain YBS01, Lactobacillus plantarum strain ATCC8014, and Saccharomyces cerevisiae strain ATCC9763 with a live bacteria count ratio of 10:20:1:1, and the total live bacteria count is 2×10⁻⁶. 10 CFU / g.

[0050] The preparation method is the same as in Example 1.

[0051] Example 7

[0052] This embodiment provides a microbial agent with deodorizing effect, comprising Bacillus megaterium strain YBG02, Bacillus subtilis strain YBS01, and Saccharomyces cerevisiae strain ATCC9763 with a live bacteria ratio of 10:10:1, and a total live bacteria count of 2 × 10⁻⁶. 10 CFU / g.

[0053] The preparation method is the same as in Example 1.

[0054] Example 8

[0055] This embodiment provides a microbial agent with deodorizing effect, comprising Bacillus megaterium strain YBG02, Bacillus subtilis strain YBS01, and Lactobacillus plantarum strain ATCC8014 with a live bacteria count ratio of 10:10:1, and a total live bacteria count of 2 × 10⁻⁶. 10 CFU / g.

[0056] The preparation method is the same as in Example 1.

[0057] Comparative Example 1

[0058] This comparative example provides a microbial agent with deodorizing effect, the microbial agent comprising Bacillus megaterium strain YBG02, with a total viable count of 2 × 10⁻⁶. 10 CFU / g.

[0059] The preparation method is the same as in Example 1.

[0060] Comparative Example 2

[0061] This comparative example provides a microbial agent with deodorizing effect, the microbial agent comprising Bacillus subtilis strain YBS01, with a total viable count of 2 × 10⁻⁶. 10 CFU / g.

[0062] The preparation method is the same as in Example 1.

[0063] Comparative Example 3

[0064] This comparative example provides a microbial agent with deodorizing effect. The only difference between this example and Example 1 is that the number of live bacteria such as "Bacillus subtilis YBS01 strain" is replaced with "Bacillus subtilis ATCC2233 strain", while the rest remains unchanged.

[0065] The preparation method is the same as in Example 1.

[0066] Comparative Example 4

[0067] This comparative example provides a microbial agent with deodorizing effect. The only difference between this example and Example 1 is that the viable bacteria such as "Bacillus megaterium YBG02 strain" are replaced with "Bacillus megaterium ATCC9885 strain", while all other aspects remain unchanged.

[0068] The preparation method is the same as in Example 1.

[0069] Application Example 1

[0070] This application example provides a cat litter with deodorizing effect. The cat litter comprises, by weight, 50 parts pea residue, 50 parts corn starch, and 1 part guar gum, and by live bacteria, 2 × 10⁻⁶ of the deodorizing microbial agent described in Example 1. 7 CFU / g.

[0071] Its preparation method is as follows:

[0072] The raw materials are physically mixed, the moisture content is adjusted to 40%, and the mixture is extruded and granulated at 80°C. The extruded and granulated cat litter is then dried at 120°C until the moisture content is 12%.

[0073] Application Example 2

[0074] This application example provides a cat litter with deodorizing effect. The cat litter comprises, by weight, 30 parts pea residue, 40 parts corn starch, and 5 parts guar gum, and by live bacteria, 2 × 10⁻⁶ of the deodorizing microbial agent described in Example 2. 7 CFU / g.

[0075] The preparation method is the same as in Example 1.

[0076] Application Example 3

[0077] This application example provides a cat litter with deodorizing effect. The cat litter comprises, by weight, 40 parts pea residue, 20 parts corn starch, and 3 parts guar gum, and by live bacteria, 2 × 10⁻⁶ of the deodorizing microbial agent described in Example 3. 7 CFU / g.

[0078] The preparation method is the same as in Example 1.

[0079] Application Example 4-8

[0080] This application example provides five types of cat litter with deodorizing effects. The only difference between this application example and application example 1 is that the number of live bacteria, such as the deodorizing agent described in example 1, is replaced with the deodorizing agent described in examples 4-8. All other ingredients and contents remain unchanged.

[0081] The preparation method is described in Application Example 1.

[0082] Compare and contrast examples 1-4

[0083] This comparative application example provides four types of cat litter with deodorizing effects. The only difference between this example and application example 1 is that the number of live bacteria, such as the deodorizing agent described in example 1, is replaced with the deodorizing agent described in comparative examples 1-4. All other components and contents remain unchanged.

[0084] The preparation method is described in Application Example 1.

[0085] Test Example 1

[0086] Stability testing

[0087] Test method:

[0088] Take 10g of each cat litter sample and grind it thoroughly in a mortar. Take 1.00g of each sample and place it in a stoppered sterile test tube. Add 10mL of sterile water and shake for 30min. Dilute the sample with sterile water to seven gradients (each gradient is diluted 10 times). Detect the bacterial count at the 5th, 6th, and 7th gradients (counting the colony count in the plate from 30 to 300). Then, place the packaged bioactive cat litter in a 37℃ oven for accelerated testing. Detect the bacterial count using the same method on days 7, 14, and 28.

[0089] Table 1

[0090]

[0091] As shown in Table 1, the strains in the cat litter samples prepared by this invention have strong stability and can maintain a high number of viable bacteria within 28 days.

[0092] Test Example 2

[0093] Evaluation of odor control ability in actual cat ownership

[0094] Test samples: Application Examples 1-8, Comparative Application Examples 1-4

[0095] 1. Sensory evaluation

[0096] 1.1 Test Method: Litter boxes containing different test samples were placed in the same cage. Each litter box contained 6L of litter. Three female cats (Golden Shaded) were housed for 48 hours. 100g of fresh feces and 400g of clumped litter from the litter boxes were collected in a shallow dish. Pet urine pads were cut into 10cm x 10cm pieces and suspended above the newly clumped litter pads for 30 minutes to fully absorb the odors emitted by the feces and urine. These pieces were collected and stored as odor sources. The litter samples were labeled accordingly. After 10 minutes, six researchers with a keen sense of smell conducted a sensory evaluation of the odor from a distance of 20cm above the urine pads.

[0097] Table 2

[0098]

[0099] Table 3

[0100]

[0101] As shown in Table 3, the cat litter containing the deodorizing bacteria agent described in this invention has a significant deodorizing effect, and Bacillus megaterium and Bacillus subtilis have a certain degree of synergy in the above-mentioned effects.

[0102] 2. Evaluation of ammonia release from collected cat feces and litter clumps containing urine.

[0103] Different application examples of clumped cat litter mixed with cat urine and fresh feces were mixed at a ratio of 6:1. 100 g of each mixture was placed in a 500 mL Erlenmeyer flask, manually shaken to mix thoroughly, and the flask opening was sealed with four layers of gauze to maintain some air permeability. The flask was placed in an artificial climate chamber with a temperature of 25℃, humidity of 30%, and light intensity of 2000 lx for 12 hours daily. Ammonia release (upper limit 500 ppm) was measured simultaneously on days 1-11 using a portable odor detector: INSCO (model VENTIS Pro5). The results are shown in Table 4.

[0104] Table 4

[0105]

[0106] As shown in Table 4, cat litter containing the deodorizing bacteria agent described in this invention can reduce the release of ammonia. Furthermore, strains YBG02 and YBS01 have a certain synergistic effect on the above-mentioned effects. After adding strains ATCC8014 and ATCC9763, the release of ammonia can be further reduced.

[0107] 3. Evaluation of hydrogen sulfide release from collected cat feces and litter clumps containing urine.

[0108] 3.1 Test Method:

[0109] Different application examples of clumped cat litter mixed with pet cat urine and fresh feces were mixed at a ratio of 6:1. 100g of each mixture was placed in a 500mL Erlenmeyer flask, manually shaken to mix thoroughly, and the flask opening was sealed with four layers of gauze to maintain some air permeability. The flask was placed in an artificial climate chamber with a temperature of 25℃, humidity of 30%, and light intensity of 2000lx for 12 hours daily. The hydrogen sulfide release was measured simultaneously on days 1-11 using a portable odor detector (VENTIS Pro5, USA). The results are shown in Table 5.

[0110] Table 5

[0111]

[0112] As shown in Table 5, cat litter containing the deodorizing bacteria agent described in this invention can reduce the release of hydrogen sulfide. Furthermore, strains YBG02 and YBS01 have a certain synergistic effect on the above-mentioned effects. Adding strains ATCC8014 and ATCC9763 can further reduce the release of hydrogen sulfide.

[0113] The applicant declares that this invention illustrates a deodorizing microbial agent, its preparation method, and its application through the above embodiments. However, this invention is not limited to the above embodiments, meaning that this invention does not necessarily rely on the above embodiments for implementation. Those skilled in the art should understand that any improvements to this invention, equivalent substitutions of raw materials, additions of auxiliary components, and selection of specific methods, etc., all fall within the protection and disclosure scope of this invention.

[0114] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0115] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not describe the various possible combinations separately.

Claims

1. A microbial agent with deodorizing effect, characterized in that, The bacterial strain in the inoculum is Bacillus megaterium with accession number CCTCC NO: M2021052. Bacillus megaterium Bacillus subtilis strain YBG02, preservation number CCTCC NO: M2021053 Bacillus subtilis YBS01 strain, Lactobacillus plantarum Lactiplantibacillus Plantarum ATCC8014 strain and Saccharomyces cerevisiae Saccharomyces cerevisiae Composition of ATCC9763 strain; The Bacillus megaterium Bacillus megaterium YBG02 strain, Bacillus subtilis Bacillus subtilis YBS01 strain, Lactobacillus plantarum Lactiplantibacillus Plantarum ATCC8014 strain and Saccharomyces cerevisiae Saccharomyces cerevisiae The ratio of viable cells in strain ATCC9763 was (10-20):(10-20):1:

1.

2. The deodorizing agent according to claim 1, characterized in that, The viable count of the bacterial agent is not less than 2 × 10⁻⁶. 10 CFU / mL or 2×10 10 CFU / g.

3. A method for preparing a microbial agent with deodorizing effect according to claim 1 or 2, characterized in that, The preparation method includes: activating the bacterial strain, fermenting it and mixing it with a protectant, freeze-drying it, and mixing the resulting bacterial powder with a carrier to obtain the final product.

4. The application of a deodorizing microbial agent according to claim 1 or 2 in the preparation of deodorizing cat litter.

5. A cat litter with deodorizing effect, characterized in that, The raw materials for preparing the cat litter include pea residue, corn starch, guar gum, and the deodorizing agent as described in claim 1 or 2.

6. The cat litter with deodorizing effect according to claim 5, characterized in that, The cat litter comprises, by weight, 30-50 parts pea residue, 20-50 parts corn starch, and 1-5 parts guar gum, and by live bacteria, 1×10⁻⁶ of the deodorizing microbial agent as described in claim 1 or 2. 7 CFU / g-2×10 7 CFU / g.

7. A method for preparing cat litter with deodorizing effect according to claim 5 or 6, characterized in that, The preparation method includes: physically mixing the raw materials, granulating, and drying to obtain the final product.

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