CN114797442B - Composite biological deodorant for animal houses and preparation method thereof - Google Patents

Composite biological deodorant for animal houses and preparation method thereof Download PDF

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CN114797442B
CN114797442B CN202210292443.7A CN202210292443A CN114797442B CN 114797442 B CN114797442 B CN 114797442B CN 202210292443 A CN202210292443 A CN 202210292443A CN 114797442 B CN114797442 B CN 114797442B
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pseudomonas
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贾银宝
岳彦
李炜
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Inner Mongolia Silong Biotechnology Co ltd
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Abstract

The invention discloses a compound biological deodorant for animal houses and a preparation method thereof, which relates to the environmental bioengineering technologyThe field of surgery. The invention provides a composite animal shed biological deodorant, which is prepared by respectively carrying out liquid fermentation on bacillus cereus, lactobacillus paracasei, pseudomonas, candida krusei, lysine bacillus and alcaligenes faecalis, then mixing to prepare mixed bacterial liquid, then adding plant essential oil mixed milk, and fully mixing, wherein the finished deodorant has obvious deodorizing effect on NH (NH) 3 The removal rate is more than or equal to 77.59 percent, H 2 The S removing rate is more than or equal to 74.27%, the malodor removing rate is more than or equal to 81.52%, the fly expelling rate is more than or equal to 81.25%, the preparation method of the product is simple, the process flow is short, the cost is low, the use is convenient, and the product is a green, environment-friendly, safe and efficient composite microbial deodorizing product.

Description

Composite biological deodorant for animal houses and preparation method thereof
Technical Field
The invention relates to the technical field of environmental bioengineering, in particular to a composite biological deodorant for animal houses and a preparation method thereof.
Background
Along with the continuous improvement of the large-scale and intensive degree of livestock industry production in China, the pollution of malodorous gas generated in the livestock production and resource utilization process to the environment is continuously increased, and the malodorous gas with complex components mainly comprises ammonia, hydrogen sulfide, indole and derivatives thereof and the like. The malodor pollution not only jeopardizes the health and production performance of livestock and poultry, but also affects the normal production and life of human beings. Therefore, the odor pollution of farms is getting more and more attention. At present, the development and use of deodorants have become a hotspot of research, except through reasonable nutritional formulas, scientific farm planning design and feeding management.
At present, the method for removing the odor mainly comprises the following steps: chemical deodorization methods such as chemical oxidation, washing-adsorption, adsorption-oxidation, etc.; physical deodorization methods such as a mask method, a dilution diffusion method, and the like; biological deodorization is mainly carried out by deodorizing by microorganism and decomposing and converting substances with odor by physiological metabolism of microorganism. Compared with other two deodorization methods, the biological deodorization method has the advantages of high odor removal rate, low equipment operation and maintenance cost, environmental protection, no secondary pollution, low treatment cost, low energy consumption, strong pertinence, wide application range and the like. However, the existing biological deodorization method still has many problems such as incomplete strains, low viable count, slow effect and single treatment function. At present, any single deodorizing method is difficult to achieve the ideal deodorizing effect.
Disclosure of Invention
Aiming at the common defects and shortcomings in the prior art, the invention creatively provides a compound biological deodorant for animal houses and a preparation method thereof.
In a first aspect, the embodiment of the invention provides a composite animal and livestock shed biological deodorant, which comprises mixed bacteria liquid and plant essential oil mixed emulsion containing 1.0-3.0% (V/V).
Wherein, the mixed bacterial liquid comprises the following components in parts by mass: 15-30 parts of bacillus cereus, 15-30 parts of lactobacillus paracasei, 10-20 parts of pseudomonas, 10-20 parts of candida krusei, 10-20 parts of bacillus lysine and 10-20 parts of alcaligenes faecalis; the plant essential oil mixed emulsion contains camphor oil, peppermint oil and citronella oil.
In the embodiment, bacillus cereus, lactobacillus paracasei, pseudomonas, candida krusei, lysine bacillus and alcaligenes faecalis are selected to be respectively subjected to liquid fermentation and then mixed to prepare mixed bacterial liquid, then plant essential oil mixed milk is added and fully mixed to prepare the deodorant, and the deodorant has obvious deodorizing effect on NH 3 The removal rate is more than or equal to 77.59 percent, H 2 The S removing rate is more than or equal to 74.27%, the malodor removing rate is more than or equal to 81.52%, the fly expelling rate is more than or equal to 81.25%, the preparation method of the product is simple, the process flow is short, the cost is low, the use is convenient, and the product is a green, environment-friendly, safe and efficient composite microbial deodorizing product.
Wherein, bacillus cereus and lactobacillus paracasei can secrete extracellular enzyme, decompose ammonia gas and hydrogen sulfide to prevent other bacteria from growing, and degrade odorous objects; the pseudomonas can remove nitrogen in the environment and degrade organic pollutants in the environment through the denitrification of the iso-oxygen and the denitrification of the aerobic bacteria; the candida krusei can ferment to produce ethyl acetate and flat peach flavor; the lysine bacillus and the alcaligenes faecalis can degrade indole and derivatives thereof in faeces, so that malodor concentration in animal houses can be effectively reduced; the mixed emulsion of the camphor oil, the peppermint oil and the citronella oil has the functions of expelling flies, inhibiting bacteria, killing insects and covering bad smell.
The product of the invention consists of a physical deodorant capable of fast acting and a biological deodorant capable of continuously deodorizing, which complement each other, and can effectively degrade ammonia gas and hydrogen sulfide gas in the animal house, effectively reduce the concentration of ammonia gas and hydrogen sulfide gas in the animal house, degrade and remove indole and derivatives thereof in the animal house, and reduce malodor concentration in the animal house; meanwhile, the growth and reproduction of harmful bacteria can be inhibited, nitrogen and organic pollutants in the environment can be effectively removed, and the multifunctional compound feed additive has multiple functions of denitrification, fly and insect expelling, bacteriostasis, insect killing, bad smell masking and the like.
With reference to the first aspect, in a first possible embodiment of the first aspect, bacillus cereus is deposited with the China industry microbiological culture collection center (CICC, address: jiujingxian Jiuqiao Jiuzhonglu 24, mitsui, beijing, chapter-a Jiujiujiuqiao, code: 100015) on 9/8/2020, and the deposit number is 24945; lactobacillus paracasei is preserved in China center for type culture collection (CICC) for 9 months and 8 days in 2020, and the preservation number is 24946, wherein address is 24 th yard of Jiuxianqiao Jiuqian Jiuzhou Jiujiu, beijing City; pseudomonas is preserved in China center for type culture collection (CICC) for industrial microorganisms, address: narcissus Jiuxian Jiuqiong Jiuzhonglu No. 24, post code: 100015, of Beijing, and accession number 25068; the candida krusei is preserved in China center for type culture collection (CICC) for industrial microorganisms in 1 month and 1 day of 1953, address: 24 th national institute of wine, the Narcissus district, beijing city, and mail code: 100015), and the preservation number is 1273; the lysine bacillus is preserved in China center for type culture collection (CICC) for industrial microorganism (address: beijing, chaoyang Jiuxian Jiuqiao Zhonglu No. 24, post code: 100015) at 12/23 of 2006, and the preservation number is 20750; alcaligenes faecalis is preserved in China industry microbiological culture Collection center (CICC for short, address: narcissus Jiuxianqiao Zhonglu 24 of Beijing City, post code: 100015) at the month of 2008, and the preservation number is 23439.
In the embodiment, the selected strains can better ensure the deodorizing performance of the product.
With reference to the first aspect, in a second possible embodiment of the first aspect, the mixed bacterial liquid contains 3.0% (V/V) of plant essential oil mixed milk, wherein the mass parts of each bacterial liquid in the mixed bacterial liquid are as follows: 25 parts of bacillus cereus, 25 parts of lactobacillus paracasei, 15 parts of pseudomonas, 15 parts of candida krusei, 10 parts of bacillus lysine and 10 parts of alcaligenes faecalis; the plant essential oil mixed emulsion comprises the following components in parts by mass: 10 parts of camphor oil, 10 parts of peppermint oil, 10 parts of citronella oil, 10 parts of sucrose fatty acid ester, 35 parts of benzyl gloss and 50 parts of water.
In the embodiment, the mixture ratio of the components of the mixed bacterial liquid and the plant essential oil mixed milk can achieve better deodorizing effect.
In a second aspect, embodiments of the present invention also provide a method for preparing a composite animal shed bio-deodorant comprising the first aspect and various possible embodiments of the first aspect.
The method comprises the following steps:
s1, respectively carrying out liquid fermentation on bacterial liquids of bacillus cereus, lactobacillus paracasei, pseudomonas, candida krusei, lysine bacillus and alcaligenes faecalis to obtain fermentation liquids;
s2, mixing the fermentation liquor obtained in the step S1 according to the following parts by weight: 15-30 parts of bacillus cereus, 15-30 parts of lactobacillus paracasei, 10-20 parts of pseudomonas, 10-20 parts of candida krusei, 10-20 parts of bacillus lysine and 10-20 parts of alcaligenes faecalis to prepare mixed bacterial liquid;
s3, mixing 5-10 parts of camphor oil, 5-10 parts of peppermint oil, 5-10 parts of citronella oil, 5-10 parts of sucrose fatty acid ester, 35-10 parts of benzyl gloss and 50-80 parts of water, and homogenizing and emulsifying for 3-10 min at the temperature of 40-70 ℃ and the rotating speed of 3000-5000 r/min by adopting a high-shear emulsifying machine to obtain plant essential oil mixed emulsion;
s4, adding 1.0-3.0% (V/V) of plant essential oil mixed emulsion into the mixed bacterial liquid, and fully mixing for 1-2 h to obtain a finished product, wherein the speed is 100-150 r/min.
In the embodiment, the product obtained by the method has more comprehensive strain functions, wherein the product is aliveThe bacterial number can reach: bacillus cereus (CFU/g) is not less than 1.43X10 8 Lactobacillus paracasei (CFU/g) is not less than 1.35X10 8 Pseudomonas (CFU/g) is more than or equal to 1.14X10 8 Candida krusei (CFU/g) is not less than 1.55X10 8 The lysine bacillus (CFU/g) is more than or equal to 1.07 multiplied by 10 8 Alcaligenes faecalis (CFU/g) is more than or equal to 1.13 multiplied by 10 8 The bacteria and a large amount of beneficial bacteria can effectively degrade ammonia gas and hydrogen sulfide gas in the livestock house in the deodorizing process, reduce the malodor concentration in the livestock house, inhibit the growth and propagation of harmful bacteria, and play a role in continuous deodorizing.
With reference to the second aspect, in a first possible embodiment of the second aspect, in S3, the homogenizing emulsifying is performed for 5min at a temperature of 50 ℃ and a rotation speed of 4000 r/min.
In the embodiment, the plant essential oil mixed emulsion prepared under the emulsification condition has lasting fragrance, is nontoxic and harmless, and does not affect livestock and environment.
With reference to the second aspect, in a second possible embodiment of the second aspect, the mixing condition in S4 is set to 120r/min, and the mixture is fully mixed for 1.5h.
In the embodiment, the mixing condition can enable the mixed bacterial liquid and the plant essential oil mixed milk to be mixed more uniformly and fully, so that the performance of the finished product is better.
Detailed Description
The following description of the technical solutions in the embodiments of the present invention will be clear and complete, and it is obvious that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The species used in the examples: bacillus cereus (Bacillus cereus), lactobacillus paracasei (Lactobacillus paracasei), pseudomonas sp, candida krusei (Candida krusei), bacillus lysiani sp. And Alcaligenes faecalis Alcaligenes faecalis used in the following examples of the present invention were all from the China center for type culture Collection of microorganisms (CICC), and were each numbered 24945, 24946, 25068, 1273 and 23439.
Example 1
1. Liquid fermentation of Bacillus cereus, lactobacillus paracasei, pseudomonas, candida krusei, bacillus lysine and Alcaligenes faecalis is carried out by the following method:
(1) Bacillus cereus fermentation broth was prepared as follows:
A. seed culture medium preparation: mixing peptone 8g, yeast extract 3g, glucose 3g, naCl8g and MnSO 4 ·H 2 Adding 0.002g of O into 1000mL of distilled water, stirring until the O is completely dissolved, sterilizing at 121 ℃ for 30min at pH of 6.5;
B. preparing a liquid fermentation medium: 8kg of peptone, 3kg of yeast extract powder, 3kg of glucose, 8kg of NaCl and MnSO 4 ·H 2 Adding 0.002kg of O into 1000L of purified water, stirring to dissolve completely, sterilizing at 121deg.C for 30min at pH 6.5;
C. culture conditions:
seed liquid culture: inoculating the inclined plane strain of bacillus cereus into a liquid seed culture medium according to the proportion of 1 loop/50 ml, and culturing for 20h at 35 ℃ at the rotating speed of 150r/min in an oscillating way;
culturing fermentation liquor: inoculating the seed liquid into a fermentation tank containing a liquid fermentation medium according to a volume ratio of 5%, and culturing for 20h at 35 ℃ at a rotational speed of 150r/min under a tank pressure of 0.02MPa at a ventilation ratio of 0.5 (V/V.min) to obtain bacillus cereus fermentation liquid.
(2) The lactobacillus paracasei fermentation broth is prepared by the following method:
A. seed culture medium preparation: mixing casein peptone 8g, beef extract 8g, yeast extract 3g, glucose 3g, sodium acetate 3g, diammonium citrate 1g, tween 80 1g, K 2 HPO 4 1g、MgSO 4 .7H 2 O 0.1g、MnSO 4 .H 2 O 0.03g、CaCO 3 15g is added into 1000mL distilled water, stirred until the mixture is completely dissolved, the pH value is 6.5, and sterilized for 30min at the temperature of 121 ℃;
B. preparing a liquid fermentation medium: casein peptone 8kg and beef extract8kg, yeast extract 3kg, glucose 3kg, sodium acetate 3kg, diammonium citrate 1kg, tween 80 1kg, K 2 HPO 4 1kg、MgSO 4 .7H 2 O 0.1kg、MnSO 4 .H 2 O 0.03kg、CaCO 3 15kg is added into 1000L purified water, stirred until the purified water is completely dissolved, the pH value is 6.5, and sterilized for 30min at the temperature of 121 ℃;
C. culture conditions:
seed liquid culture: inoculating lactobacillus paracasei inclined plane strain into a liquid seed culture medium according to the proportion of 1 loop/50 ml, and performing shake culture for 20h at 35 ℃ at the rotating speed of 100 r/min;
culturing fermentation liquor: inoculating the seed liquid into a fermentation tank containing a liquid fermentation medium according to a volume ratio of 5%, and culturing for 20h at 35 ℃ at a speed of 100r/min at a tank pressure of 0.02MPa and a ventilation ratio of 0.5 (V/V.min) to obtain lactobacillus paracasei fermentation liquid.
(3) The Pseudomonas fermentation broth is prepared as follows:
A. seed culture medium preparation: adding 8g of peptone, 3g of yeast extract powder, 3g of glucose and 8g of NaCl into 1000mL of distilled water, stirring until the peptone, the yeast extract powder and the NaCl are completely dissolved, sterilizing at 121 ℃ for 30min;
B. preparing a liquid fermentation medium: adding peptone 8kg, yeast extract 3kg, glucose 3kg, and NaCl 8kg into 1000L purified water, stirring to dissolve completely, sterilizing at 121deg.C for 30min at pH 6.5;
C. culture conditions:
seed liquid culture: inoculating Pseudomonas inclined plane strain into liquid seed culture medium at 28deg.C at 150r/min at a ratio of 1 ring/50 ml, and shake culturing for 20 hr;
culturing fermentation liquor: inoculating the seed solution into a fermentation tank containing a liquid fermentation medium according to a volume ratio of 5%, and culturing at 28 ℃ at a ventilation ratio of 0.5 (V/V.min), a tank pressure of 0.02MPa and a rotating speed of 150r/min for 20h to obtain the pseudomonas fermentation broth.
(4) The candida krusei fermentation broth is prepared by the following method:
A. seed culture medium preparation: adding 5g of malt extract powder, 1g of maltose, 1g of yeast extract powder and 5g of glucose into 1000mL of distilled water, stirring until the materials are completely dissolved, sterilizing at 115 ℃ for 30min, wherein the pH is 4.5;
B. preparing a liquid fermentation medium: adding malt extract 5kg, maltose 1kg, yeast extract 1kg, glucose 5kg into 1000L distilled water, stirring to dissolve completely, sterilizing at 115deg.C for 30min;
C. culture conditions:
seed liquid culture: inoculating the candida krusei inclined plane strain into a liquid seed culture medium according to the proportion of 1 ring/50, and carrying out shake culture for 20h at the temperature of 28 ℃ and the rotating speed of 150 r/min;
culturing fermentation liquor: inoculating the seed liquid into a fermentation tank containing a liquid fermentation medium according to a volume ratio of 5%, and culturing for 20h at 28 ℃ at a rotation speed of 150r/min under a tank pressure of 0.02MPa at a ventilation ratio of 0.5 (V/V.min) to obtain the candida krusei fermentation liquid.
(5) The lysine bacillus fermentation broth is prepared as follows:
A. seed culture medium preparation: adding 8g of peptone, 3g of yeast extract powder, 3g of glucose and 8g of NaCl into 1000mL of distilled water, stirring until the peptone, the yeast extract powder and the NaCl are completely dissolved, sterilizing at 121 ℃ for 30min;
B. preparing a liquid fermentation medium: adding peptone 8kg, yeast extract 3kg, glucose 3kg, and NaCl 8kg into 1000L purified water, stirring to dissolve completely, sterilizing at 121deg.C for 30min at pH 6.5;
C. culture conditions:
seed liquid culture: inoculating lysine bacillus slant strain into liquid seed culture medium at 35 deg.C and rotation speed of 150r/min at a ratio of 1 loop/50 ml, and shake culturing for 20 hr;
culturing fermentation liquor: inoculating the seed liquid into a fermentation tank containing a liquid fermentation medium according to a volume ratio of 5%, and performing shake culture for 20h at 35 ℃ at a pressure of 0.02MPa and a rotation speed of 150r/min at a ventilation ratio of 0.5 (V/V.min) to obtain the lysine bacillus fermentation liquid.
(6) The alcaligenes faecalis fermentation broth is prepared according to the following method:
A. seed culture medium preparation: adding 8g of peptone, 2g of beef extract powder and 3g of NaCl into 1000mL of distilled water, stirring until the peptone, the beef extract powder and the NaCl are completely dissolved, sterilizing at 121 ℃ for 30min;
B. preparing a liquid fermentation medium: adding peptone 8kg, beef extract powder 2kg and NaCl3kg into 1000L purified water, stirring to dissolve completely, sterilizing at 121deg.C for 30min;
C. culture conditions:
seed liquid culture: inoculating the alcaligenes faecalis slant strain into a liquid seed culture medium according to the ratio of 1 loop/50, and performing shake culture for 20h at 28 ℃ at the rotating speed of 150 r/min;
culturing fermentation liquor: inoculating the seed liquid into a fermentation tank containing a liquid fermentation medium according to a volume ratio of 5%, and performing shake culture for 20h at 28 ℃ at a pressure of 0.02MPa and a rotation speed of 150r/min at a ventilation ratio of 0.5 (V/V.min) to obtain the alcaligenes faecalis fermentation liquid.
2. The prepared bacillus cereus, lactobacillus paracasei, pseudomonas, candida krusei, lysine bacillus and alcaligenes faecalis bacterial solutions are respectively compounded according to the following proportion (mass portion):
15 parts of bacillus cereus, 15 parts of lactobacillus paracasei, 20 parts of pseudomonas, 20 parts of candida krusei, 15 parts of lysinibacillus and 15 parts of alcaligenes faecalis to obtain a first mixed bacterial liquid.
3. The camphor oil, the peppermint oil, the citronella oil, the emulsifier and the water are compounded according to the following proportion (mass portion) and are prepared according to the following method:
5 parts of camphor oil, 5 parts of peppermint oil, 5 parts of citronella oil, 5 parts of sucrose fatty acid ester, 35 parts of benzyl gloss and 75 parts of water, and homogenizing and emulsifying for 5min at 50 ℃ at the rotating speed of 4000r/min by a high-shear emulsifying machine to obtain the first plant essential oil mixed emulsion.
4. And finally, adding the first plant essential oil mixed emulsion into the first mixed bacterial liquid according to the proportion of 1.0% (V/V), and fully mixing for 1.5h at 120r/min to obtain a finished product, wherein the finished product is marked as a test group I.
Example two
1. Liquid fermentation of Bacillus cereus, lactobacillus paracasei, pseudomonas, candida krusei, bacillus lysine and Alcaligenes faecalis is carried out by the following method:
(1) Bacillus cereus fermentation broth was prepared as follows:
A. seed culture medium preparation: 10g of peptone, 5g of yeast extract powder, 5g of glucose, 10g of NaCl and MnSO 4 ·H 2 Adding 0.004g of O into 1000mL of distilled water, stirring until the O is completely dissolved, and sterilizing at 121 ℃ for 30min;
B. preparing a liquid fermentation medium: 10kg of peptone, 5kg of yeast extract powder, 5kg of glucose, 10kg of NaCl and MnSO 4 ·H 2 Adding 0.004kg of O into 1000L of purified water, stirring until the O is completely dissolved, and sterilizing at 121 ℃ for 30min;
C. culture conditions:
seed liquid culture: inoculating the inclined plane strain of bacillus cereus into a liquid seed culture medium according to the proportion of 1 loop/50 ml, and culturing for 24 hours at 37 ℃ at the rotating speed of 180r/min in an oscillating way;
culturing fermentation liquor: inoculating the seed liquid into a fermentation tank containing a liquid fermentation medium according to the volume ratio of 10%, and culturing for 24 hours at 37 ℃ at the rotational speed of 180r/min under the pressure of 0.03MPa at the aeration ratio of 1.0 (V/V.min) to obtain bacillus cereus fermentation liquid.
(2) The lactobacillus paracasei fermentation broth is prepared by the following method:
A. seed culture medium preparation: 10g of casein peptone, 10g of beef extract, 5g of yeast extract powder, 5g of glucose, 5g of sodium acetate, 2g of diammonium citrate, 1g of Tween 80 and K 2 HPO 4 2g、MgSO 4 .7H 2 O 0.2、MnSO 4 .H 2 O 0.05g、CaCO 3 Adding 20g into 1000mL distilled water, stirring to dissolve completely, pH7.0, and sterilizing at 121deg.C for 30min;
B. preparing a liquid fermentation medium: 10kg of casein peptone, 10kg of beef extract, 5kg of yeast extract powder, 5kg of glucose, 5kg of sodium acetate, 2kg of diammonium citrate, 1kg of Tween 80 and K 2 HPO 4 2kg、MgSO 4 .7H 2 0.2kg、MnSO 4 .H 2 O 0.05kg、CaCO 3 20kg is added into 1000L purified water, stirred until the purified water is completely dissolved, and sterilized for 30min at the temperature of 121 ℃;
C. culture conditions:
seed liquid culture: inoculating lactobacillus paracasei inclined plane strain into a liquid seed culture medium according to the proportion of 1 loop/50, and carrying out shake culture for 240h at 37 ℃ at the rotating speed of 120 r/min;
culturing fermentation liquor: inoculating the seed liquid into a fermentation tank containing a liquid fermentation medium according to the volume ratio of 10%, and culturing for 24 hours at 37 ℃ at the rotational speed of 120r/min under the pressure of 0.03MPa at the aeration ratio of 1.0 (V/V.min) to obtain lactobacillus paracasei fermentation liquor.
(3) The Pseudomonas fermentation broth is prepared as follows:
A. seed culture medium preparation: adding 10g of peptone, 5g of yeast extract powder, 5g of glucose and 10g of NaCl into 1000mL of distilled water, stirring until the peptone, the yeast extract powder and the NaCl are completely dissolved, and sterilizing at 121 ℃ for 30min;
B. preparing a liquid fermentation medium: adding peptone 10kg, yeast extract 5kg, glucose 5kg, and NaCl 10kg into 1000L purified water, stirring to dissolve completely, pH7.0, and sterilizing at 121deg.C for 30min;
C. culture conditions:
seed liquid culture: inoculating Pseudomonas inclined plane strain into liquid seed culture medium at 30deg.C at 180r/min at a ratio of 1 ring/50 ml, and shake culturing for 24 hr;
culturing fermentation liquor: inoculating the seed solution into a fermentation tank containing a liquid fermentation medium according to the volume ratio of 10%, and culturing for 24 hours at the temperature of 30 ℃ and the aeration ratio of 1.0 (V/V.min), the tank pressure of 0.03MPa and the rotating speed of 180r/min to obtain the pseudomonas fermentation liquid.
(4) The candida krusei fermentation broth is prepared by the following method:
A. seed culture medium preparation: adding 8g of malt extract powder, 1.8g of maltose, 1.6g of yeast extract powder and 6g of glucose into 1000mL of distilled water, stirring until the materials are completely dissolved, sterilizing at 115 ℃ for 30min, wherein the pH is 5.0;
B. preparing a liquid fermentation medium: adding malt extract 8kg, maltose 1.8kg, yeast extract 1.6kg, and glucose 6kg into 1000L distilled water, stirring to dissolve completely, sterilizing at 115 deg.C for 30min;
C. culture conditions:
seed liquid culture: inoculating the candida krusei inclined plane strain into a liquid seed culture medium according to the proportion of 1 loop/50 ml, and carrying out shake culture for 24 hours at the temperature of 30 ℃ and the rotating speed of 180 r/min;
culturing fermentation liquor: inoculating the seed liquid into a fermentation tank containing a liquid fermentation medium according to the volume ratio of 10%, and culturing for 24 hours at the temperature of 30 ℃ and the aeration ratio of 1.0 (V/V.min), the tank pressure of 0.03MPa and the rotating speed of 180r/min to obtain the candida krusei fermentation liquid.
(5) The lysine bacillus fermentation broth is prepared as follows:
A. seed culture medium preparation: adding 10g of peptone, 5g of yeast extract powder, 5g of glucose and 10g of NaCl into 1000mL of distilled water, stirring until the peptone, the yeast extract powder and the NaCl are completely dissolved, and sterilizing at 121 ℃ for 30min;
B. preparing a liquid fermentation medium: adding peptone 10kg, yeast extract 5kg, glucose 5kg, and NaCl 10kg into 1000L purified water, stirring to dissolve completely, pH7.0, and sterilizing at 121deg.C for 30min;
C. culture conditions:
seed liquid culture: inoculating lysine bacillus slant strain into liquid seed culture medium at 37 deg.c and rotation speed of 180r/min in the ratio of 1 loop/50 ml, and oscillating culture for 24 hr;
culturing fermentation liquor: inoculating the seed solution into a fermentation tank containing a liquid fermentation medium according to the volume ratio of 10%, and performing shaking culture for 24 hours at 37 ℃ at the rotational speed of 180r/min under the pressure of 0.03 and the aeration ratio of 1.0 (V/V.min) to obtain the lysine bacillus fermentation liquid.
(6) The alcaligenes faecalis fermentation broth is prepared according to the following method:
A. seed culture medium preparation: adding 10g of peptone, 3g of beef extract powder and 5g of NaCl into 1000mL of distilled water, stirring until the peptone, the beef extract powder and the NaCl are completely dissolved, and sterilizing at 121 ℃ for 30min;
B. preparing a liquid fermentation medium: adding peptone 10kg, beef extract 3kg and NaCl5kg into 1000L purified water, stirring to dissolve completely, sterilizing at 121deg.C for 30min;
C. culture conditions:
seed liquid culture: inoculating the alcaligenes faecalis slant strain into a liquid seed culture medium according to the proportion of 1 loop/50 ml, and culturing at 30 ℃ at the rotation speed of 180r/min for 24 hours in an oscillating way;
culturing fermentation liquor: inoculating the seed liquid into a fermentation tank containing a liquid fermentation medium according to the volume ratio of 10%, and performing shake culture for 24 hours at the temperature of 30 ℃ and the aeration ratio of 1.0 (V/V.min), the tank pressure of 0.03MPa and the rotating speed of 180r/min to obtain the alcaligenes faecalis fermentation liquid.
2. The prepared bacillus cereus, lactobacillus paracasei, pseudomonas, candida krusei, lysine bacillus and alcaligenes faecalis bacterial solutions are respectively compounded according to the following proportion (mass portion):
20 parts of bacillus cereus, 20 parts of lactobacillus paracasei, 10 parts of pseudomonas, 10 parts of candida krusei, 20 parts of lysinibacillus and 20 parts of alcaligenes faecalis to obtain a second mixed bacterial liquid.
3. The camphor oil, the peppermint oil, the citronella oil, the emulsifier and the water are compounded according to the following proportion (mass portion) and are prepared according to the following method:
8 parts of camphor oil, 8 parts of peppermint oil, 8 parts of citronella oil, 8 parts of sucrose fatty acid ester, 35 parts of benzyl gloss and 60 parts of water, and homogenizing and emulsifying for 5min at the temperature of 50 ℃ and the rotating speed of 4000r/min by adopting a high-shear emulsifying machine to obtain the second plant essential oil mixed emulsion.
5. And finally, adding the second plant essential oil mixed emulsion into the second mixed bacterial liquid according to the proportion of 2.0% (V/V), and fully mixing for 1.5h at 120r/min to obtain a finished product, wherein the finished product is marked as a test group II.
Example III
1. Liquid fermentation of Bacillus cereus, lactobacillus paracasei, pseudomonas, candida krusei, bacillus lysine and Alcaligenes faecalis is carried out by the following method:
(1) Bacillus cereus fermentation broth was prepared as follows:
A. seed culture medium preparation: 12g of peptone, 7g of yeast extract powder, 7g of glucose, 12g of NaCl and MnSO 4 ·H 2 Adding 0.006g of O into 1000mL of distilled water, stirring to dissolve completely, and sterilizing at 121deg.C for 30min;
B. preparing a liquid fermentation medium: 12kg of protein, 7kg of yeast extract powder, 7kg of glucose, 12kg of NaCl and MnSO 4 ·H 2 Adding 0.006kg of O into 1000L of purified water, stirring to dissolve completely, pH7.5, and sterilizing at 121deg.C for 30min;
C. culture conditions:
seed liquid culture: inoculating the bacillus cereus inclined plane strain into a liquid seed culture medium according to the proportion of 1 loop/50 ml, and carrying out shake culture for 30h at the temperature of 40 ℃ at the rotating speed of 200 r/min;
culturing fermentation liquor: inoculating the seed liquid into a fermentation tank containing a liquid fermentation medium according to the volume ratio of 15%, and culturing for 30 hours at 40 ℃ at the rotational speed of 200r/min under the pressure of 0.05MPa at the aeration ratio of 1.5 (V/V.min) to obtain bacillus cereus fermentation liquid.
(2) The lactobacillus paracasei fermentation broth is prepared by the following method:
A. seed culture medium preparation: 12g of casein peptone, 12g of beef extract, 7g of yeast extract powder, 7g of glucose, 7g of sodium acetate, 2g of diammonium citrate, 2g of Tween 80 and K 2 HPO 4 3g、MgSO 4 .7H 2 O 0.3g、MnSO 4 .H 2 O 0.07g、CaCO 3 25g of the mixture is added into 1000mL of distilled water, stirred until the mixture is completely dissolved, pH7.5 and sterilized for 30min at the temperature of 121 ℃;
B. preparing a liquid fermentation medium: 12kg of casein peptone, 12kg of beef extract, 7kg of yeast extract powder, 7kg of glucose, 7kg of sodium acetate, 2kg of diammonium citrate, 2kg of Tween 80 and K 2 HPO 4 3kg、MgSO 4 .7H 2 O 0.3kg、MnSO 4 .H 2 O 0.07kg、CaCO 3 25kg is added into 1000L purified water, stirred until completely dissolved, pH7.5, sterilized at 121 ℃ for 30min;
C. culture conditions:
seed liquid culture: inoculating lactobacillus paracasei inclined plane strain into a liquid seed culture medium according to the proportion of 1 loop/50 ml, and performing shake culture for 30 hours at 40 ℃ at the rotating speed of 150 r/min;
culturing fermentation liquor: inoculating the seed liquid into a fermentation tank containing a liquid fermentation medium according to the volume ratio of 15%, and culturing for 30 hours at the temperature of 40 ℃ and the aeration ratio of 1.5 (V/V.min), the tank pressure of 0.05MPa and the rotating speed of 100-150 r/min to obtain the lactobacillus paracasei fermentation liquid.
(3) The Pseudomonas fermentation broth is prepared as follows:
A. seed culture medium preparation: adding 12g of peptone, 7g of yeast extract powder, 7g of glucose and 12g of NaCl into 1000mL of distilled water, stirring until the peptone, the 7.5 g of yeast extract powder and the NaCl are completely dissolved, and sterilizing for 30min at the temperature of 121 ℃;
B. preparing a liquid fermentation medium: adding peptone 12kg, yeast extract 7kg, glucose 7kg, and NaCl 12kg into 1000L purified water, stirring to dissolve completely, pH7.5, and sterilizing at 121deg.C for 30min;
C. culture conditions:
seed liquid culture: inoculating Pseudomonas inclined plane strain into liquid seed culture medium at 35 deg.C at 200r/min at a ratio of 1 ring/50 ml, and shake culturing for 30 hr;
culturing fermentation liquor: inoculating the seed solution into a fermentation tank containing a liquid fermentation medium according to a volume ratio of 15%, and culturing for 30h at 35 ℃ at a rotational speed of 200r/min under a pressure of 0.05MPa at a ventilation ratio of 1.5 (V/V.min) to obtain the Pseudomonas fermentation broth.
(4) The candida krusei fermentation broth is prepared by the following method:
A. seed culture medium preparation: adding 10g of malt extract powder, 2g of maltose, 2g of yeast extract powder and 10g of glucose into 1000mL of distilled water, stirring until the materials are completely dissolved, sterilizing at 115 ℃ for 30min, wherein the pH is 5.5;
B. preparing a liquid fermentation medium: adding 10kg of malt extract powder, 2kg of maltose, 2kg of yeast extract powder and 10kg of glucose into 1000L of distilled water, stirring until the materials are completely dissolved, sterilizing at 115 ℃ for 30min, wherein the pH is 5.5;
C. culture conditions:
seed liquid culture: inoculating the candida krusei inclined plane strain into a liquid seed culture medium according to the proportion of 1 ring per 50ml, and carrying out shake culture for 30 hours at the temperature of 35 ℃ and the rotating speed of 200 r/min;
culturing fermentation liquor: inoculating the seed liquid into a fermentation tank containing a liquid fermentation medium according to the volume ratio of 15%, and culturing for 30 hours at 35 ℃ at the rotational speed of 200r/min under the pressure of 0.05MPa at the aeration ratio of 1.5 (V/V.min) to obtain the candida krusei fermentation liquid.
(5) The lysine bacillus fermentation broth is prepared as follows:
A. seed culture medium preparation: adding 12g of peptone, 7g of yeast extract powder, 7g of glucose and 12g of NaCl into 1000mL of distilled water, stirring until the peptone, the 7.5 g of yeast extract powder and the NaCl are completely dissolved, and sterilizing for 30min at the temperature of 121 ℃;
B. preparing a liquid fermentation medium: adding peptone 12kg, yeast extract 7kg, glucose 7kg, and NaCl 12kg into 1000L purified water, stirring to dissolve completely, pH7.5, and sterilizing at 121deg.C for 30min;
C. culture conditions:
seed liquid culture: inoculating lysine bacillus slant strain into liquid seed culture medium at 40 deg.C and rotation speed of 200r/min at a ratio of 1 loop/50 ml, and shake culturing for 30 hr;
culturing fermentation liquor: inoculating the seed solution into a fermentation tank containing a liquid fermentation medium according to a volume ratio of 15%, and performing shaking culture for 30 hours at 40 ℃ at a rotational speed of 200r/min under a tank pressure of 0.05MPa at a ventilation ratio of 1.5 (V/V.min) to obtain a lysine bacillus fermentation broth.
(6) The alcaligenes faecalis fermentation broth is prepared according to the following method:
A. seed culture medium preparation: adding 12g of peptone, 5g of beef extract powder and 7g of NaCl into 1000mL of distilled water, stirring until the peptone and the beef extract powder are completely dissolved, sterilizing at 121 ℃ for 30min;
B. preparing a liquid fermentation medium: adding peptone 12kg, beef extract 5kg and NaCl 7kg into 1000L purified water, stirring to dissolve completely, sterilizing at 121deg.C for 30min;
C. culture conditions:
seed liquid culture: inoculating the alcaligenes faecalis slant strain into a liquid seed culture medium according to the proportion of 1 loop/50 ml, and culturing at 35 ℃ at the rotation speed of 200r/min for 30h in an oscillating way;
culturing fermentation liquor: inoculating the seed liquid into a fermentation tank containing a liquid fermentation medium according to the volume ratio of 15%, and performing shake culture for 30 hours at 35 ℃ at the pressure of 0.05MPa and the rotation speed of 200r/min at the aeration ratio of 1.5 (V/V.min) to obtain the alcaligenes faecalis fermentation liquid.
2. The prepared bacillus cereus, lactobacillus paracasei, pseudomonas, candida krusei, lysine bacillus and alcaligenes faecalis bacterial solutions are respectively compounded according to the following proportion (mass portion):
25 parts of bacillus cereus, 25 parts of lactobacillus paracasei, 15 parts of pseudomonas, 15 parts of candida krusei, 10 parts of lysinibacillus and 10 parts of alcaligenes faecalis to obtain a third mixed bacterial liquid.
3. The camphor oil, the peppermint oil, the citronella oil, the emulsifier and the water are compounded according to the following proportion (mass portion) and are prepared according to the following method:
10 parts of camphor oil, 10 parts of peppermint oil, 10 parts of citronella oil, 10 parts of sucrose fatty acid ester, 35 parts of benzyl gloss and 50 parts of water, and homogenizing and emulsifying for 5min at the temperature of 50 ℃ and the rotating speed of 4000r/min by adopting a high-shear emulsifying machine to obtain the third plant essential oil mixed emulsion.
4. And finally, adding the third plant essential oil mixed emulsion into the third mixed bacterial liquid according to the proportion of 3.0% (V/V), and fully mixing for 1.5h at 120r/min to obtain a finished product, wherein the finished product is marked as a test group III.
Example IV
The viable count of the test group I, the test group II and the test group III is detected in a plate counting mode, and the measurement results are as follows:
test group i: bacillus cereus (CFU/g) is not less than 1.43X10 8 Lactobacillus paracasei (CFU/g) is not less than 1.35X10 8 Pseudomonas (CFU/g) is more than or equal to 1.48X10 8 Candida krusei (CFU/g) is not less than 1.86×10 8 The lysine bacillus (CFU/g) is more than or equal to 1.33X10 8 Alcaligenes faecalis (CFU/g) is more than or equal to 1.36 multiplied by 10 8 ；
Test group ii: bacillus cereus (CFU/g) is not less than 3.13×10 8 Lactobacillus paracasei (CFU/g) is not less than 2.06X10 8 Pseudomonas (CFU/g) is more than or equal to 1.14X10 8 Candida krusei (CFU/g) is not less than 1.55X10 8 The lysine bacillus (CFU/g) is more than or equal to 2.10X10 8 Alcaligenes faecalis (CFU/g) is more than or equal to 2.07 multiplied by 10 8 ；
Test group III: bacillus cereus (CFU/g) is not less than 3.53X10 8 Lactobacillus paracasei (CFU/g) is not less than 2.25X10 8 Pseudomonas (CFU/g) is more than or equal to 1.39X10 8 Candida krusei (CFU/g) is more than or equal to 2.12X10 8 The lysine bacillus (CFU/g) is more than or equal to 1.07 multiplied by 10 8 Alcaligenes faecalis (CFU/g) is more than or equal to 1.13 multiplied by 10 8 。
In summary, the deodorant prepared by the method of the invention has the following viable count: bacillus cereus (CFU/g) is more than or equal to 1.43×108, lactobacillus paracasei (CFU/g) is more than or equal to 1.35×108, pseudomonas (CFU/g) is more than or equal to 1.14×108, candida krusei (CFU/g) is more than or equal to 1.55×108, bacillus lysines (CFU/g) is more than or equal to 1.07×108, alcaligenes faecalis (CFU/g) is more than or equal to 1.13×108, and a large number of living bacteria can make the deodorizing effect of the product more obvious and durable.
Example five
Compound animal house biological deodorant deodorization test
1. Test materials
1.1 test article: test group I, test group II, test group III.
1.2 reagents and equipment: gas chromatograph, CD-1 atmospheric sampler, spectrophotometer, odorless bag, etc.
1.3 test field conditions
Table 1 test field conditions
Test conditions Test field
Total stock (head) 1000
Cultivation mode Cement solid floor and dry manure cleaning mode
Test house arrangement 3 independent test houses
Size of a house (m) 30×10
Body weight range (kg) 80±5
Other conditions Does not punch the ring and naturally ventilates
2. Test method
2.1 design of experiments
The original feeding management conditions are kept unchanged, and the deodorant tests are independently carried out in 3 test piggeries. Tap water for biological deodorant of compound animal houses of each group is prepared according to the following ratio of 1:10, and then uniformly spraying the mixture into a pig house by using a sprayer according to the amount of 100ml per square meter. 8 am every day: 00 and 20 pm: 00 is sprayed once each, and the test period is 10 days.
2.2 test determination
Sampling according to the unstructured exhaust gas HJ/T55-2000 'technical guidelines for the unstructured emission monitoring of atmospheric pollutants'; detecting ammonia gas by using a Nahner reagent spectrophotometry HJ 533-2009; hydrogen sulfide was detected by gas chromatography GB/T14678-1993; malodor concentration was measured using the three-point comparative malodor bag method GB/T14675-1993.
Each pigsty is provided with 3 test points along the diagonal of the house, 7 in the morning: 00. noon 13: 00. night 17:00, respectively measuring each index, and then taking the average value of the detection before and after the pig house test for data analysis. The mosquito and fly repellent ratio is calculated and analyzed according to the average number of mosquitoes and flies in unit square meter.
3. Analysis of test results
Table 2 analysis of results of pig farm test with composite animal housing biological deodorant
Figure BDA0003560778810000241
Figure BDA0003560778810000251
Note that: the same column of data is marked with a different shoulder letter, indicating a significant difference (P < 0.05), and the same indicates a insignificant difference (P > 0.05).
4. Discussion and conclusion
Tests prove that the deodorizing effects of the three groups of test products are obvious (P is smaller than 0.05), wherein the comprehensive effects of the test group III are optimal. After 10 days of the compound animal house biological deodorant of test group III is sprayed in the pig house, the NH3 removal rate in the pig house reaches 88.69%, the H2S removal rate reaches 84.14%, the malodor removal rate reaches 83.33%, and the fly and insect are reduced by 93.02%. Therefore, the compound biological deodorant for animal houses claimed by the invention has remarkable deodorizing effect and application prospect.
The above is only a preferred embodiment of the present invention, and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (5)

1. A composite biological deodorant for animal houses is characterized by comprising
The mixed bacterial liquid contains 1.0-3.0% of plant essential oil mixed emulsion with the concentration of V/V;
the mixed bacterial liquid comprises the following components in parts by mass: 15-30 parts of bacillus cereus, 15-30 parts of lactobacillus paracasei, 10-20 parts of pseudomonas, 10-20 parts of candida krusei, 10-20 parts of bacillus lysine and 10-20 parts of alcaligenes faecalis, wherein the bacillus cereus is preserved in China industry microbiological culture collection center (CICC) for 9 months and 8 days in 2020, address: narcissus bridge No. 24, post code: 100015, accession number 24945; the lactobacillus paracasei is preserved in China center for type culture collection (CICC) for 9 and 8 days in 2020, address: narcissus bridge No. 24, post code: 100015, accession number 24946; the pseudomonas is preserved in China center for type culture collection (CICC) for type number of industrial microorganisms, address: narcissus bridge No. 24, post code: 100015, accession number 25068; the candida krusei is preserved in the China center for type culture collection of Industrial microorganisms (CICC) for 1 month and 1 day in 1953, and addresses are given: narcissus bridge No. 24, post code: 100015, accession number 1273; the lysine bacillus is preserved in China industry microbiological culture collection center (CICC) for short, address: narcissus bridge No. 24, post code: 100015, accession number 20750; the alcaligenes faecalis is preserved in China industry microbiological culture collection center, called CICC for short, address: narcissus bridge No. 24, post code: 100015, accession number 23439;
the plant essential oil mixed emulsion contains camphor oil, peppermint oil and citronella oil.
2. The compound animal shed biological deodorant according to claim 1, wherein the mixed bacterial liquid contains 3.0% V/V of plant essential oil mixed milk, and the mass parts of each bacterial liquid in the mixed bacterial liquid are as follows: 25 parts of bacillus cereus, 25 parts of lactobacillus paracasei, 15 parts of pseudomonas, 15 parts of candida krusei, 10 parts of bacillus lysine and 10 parts of alcaligenes faecalis; the plant essential oil mixed emulsion comprises the following components in parts by mass: 10 parts of camphor oil, 10 parts of peppermint oil, 10 parts of citronella oil, 10 parts of sucrose fatty acid ester, 35 parts of benzyl gloss and 50 parts of water.
3. The preparation method of the composite animal housing biological deodorant according to any one of claims 1 to 2, which is characterized by comprising the following steps:
s1, respectively carrying out liquid fermentation on bacterial liquids of bacillus cereus, lactobacillus paracasei, pseudomonas, candida krusei, lysine bacillus and alcaligenes faecalis to obtain fermentation liquids;
s2, mixing the fermentation liquor obtained in the step S1 according to the following parts by weight: 15-30 parts of bacillus cereus, 15-30 parts of lactobacillus paracasei, 10-20 parts of pseudomonas, 10-20 parts of candida krusei, 10-20 parts of bacillus lysine and 10-20 parts of alcaligenes faecalis to prepare mixed bacterial liquid;
s3, mixing 5-10 parts of camphor oil, 5-10 parts of peppermint oil, 5-10 parts of citronella oil, 5-10 parts of sucrose fatty acid ester, 35-10 parts of benzyl gloss and 50-80 parts of water, and homogenizing and emulsifying for 3-10 min at the temperature of 40-70 ℃ and the rotating speed of 3000-5000 r/min by adopting a high-shear emulsifying machine to obtain plant essential oil mixed emulsion;
s4, adding 1.0-3.0% of the plant essential oil mixed emulsion with the concentration of V/V into the mixed bacterial liquid, and fully mixing for 1-2 hours at the speed of 100-150 r/min to obtain a finished product.
4. The method for preparing a composite animal housing biological deodorant according to claim 3, wherein the S3 is homogeneously emulsified for 5min at a temperature of 50 ℃ and a rotation speed of 4000 r/min.
5. The method for preparing a composite animal housing biological deodorant according to claim 3, wherein the mixing condition in S4 is 120r/min, and the mixture is thoroughly mixed for 1.5 hours.
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CN109609401A (en) * 2018-12-18 2019-04-12 山东省科学院生态研究所 A kind of complex microorganism deodorizing microorganism and the preparation method and application thereof
CN111500495A (en) * 2020-04-27 2020-08-07 巴彦淖尔市云天下电子商务股份有限公司 Composite microbial deodorant and preparation method thereof

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CN109609401A (en) * 2018-12-18 2019-04-12 山东省科学院生态研究所 A kind of complex microorganism deodorizing microorganism and the preparation method and application thereof
CN111500495A (en) * 2020-04-27 2020-08-07 巴彦淖尔市云天下电子商务股份有限公司 Composite microbial deodorant and preparation method thereof

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