CN114181856A

CN114181856A - Microbial composition for efficiently inhibiting odor production by biomass fermentation from source and application thereof

Info

Publication number: CN114181856A
Application number: CN202111496500.5A
Authority: CN
Inventors: 刘玉涛; 马海霞; 万和川; 李骅; 赵三琴; 顾家冰
Original assignee: Nanjing Agricultural University
Current assignee: Nanjing Agricultural University
Priority date: 2021-11-30
Filing date: 2021-11-30
Publication date: 2022-03-15
Anticipated expiration: 2041-11-30
Also published as: CN114181856B

Abstract

The invention provides a microbial composition for efficiently inhibiting odor generation by biomass fermentation from a source and application thereof, and relates to the technical field of gas purification. The active ingredients of the odor-inhibiting microbial inoculum comprise yeast derived from kitchen garbage, mature bacteria derived from livestock and poultry manure and nitrobacteria respectively; can ensure the biological safety of the plant, responds to the concept of environmental protection and emission reduction, and has wide application range. The invention provides a use method based on the combined microbial inoculum, which increases the infiltration of the microbial inoculum on biomass and improves the opportunity of microorganisms entering the biomass, thereby achieving the purposes of promoting the propagation of the microorganisms and inhibiting the generation of odor. The functions of all microorganisms in the complex flora have complementarity, thereby ensuring high efficiency and thoroughness.

Description

Microbial composition for efficiently inhibiting odor production by biomass fermentation from source and application thereof

Technical Field

The invention belongs to the technical field of gas purification, and particularly relates to an odor inhibiting strain combination with complementary functions and application thereof.

Background

The mass production of malodorous gases seriously affects the living environment of residents and harms the physical health of human beings. Livestock and poultry manure and kitchen garbage are two types of biomass which are easy to decompose and produce odor, the daily discharge amount of the livestock and poultry manure in China currently reaches 700 ten thousand tons, the annual total production amount is about 24 hundred million tons, and the production amounts of manure and urine are respectively about 16 million tons and 8 million tons; in the urban domestic garbage structure in China, the proportion of kitchen waste accounts for 30% -50%, the generation amount of kitchen waste in cities in China is not less than 6000 million tons every year, and the generation amount shows a continuously increasing trend. The main components of the livestock manure and the kitchen garbage are protein, sugar, fat and other organic substances, wherein the protein, amino acid and other organic substances can be subjected to decarboxylation and deamination due to the activity of microorganisms, so that a large amount of malodorous gas is generated, and the air quality and the body health of surrounding residents are seriously affected; therefore, the disposal of odor generated by decomposition of biomass is not slow.

At present, methods such as chemical absorption, oxidation, physical adsorption and shielding are mainly adopted for treating odor, so that the problems of secondary pollution, temporary solution and non-permanent solution exist, and because components in biomass are converted into odor, resources in the odor are partially consumed; in order to solve the problems, the development of a microbial agent which can effectively inhibit the odor generation of livestock and poultry manure and kitchen garbage from the source is urgently needed to meet the requirement of national green development.

Disclosure of Invention

Aiming at the problem that the decomposition and odor generation of biomass are difficult to treat at present, the invention discloses a microbial composition with a source capable of efficiently inhibiting the fermentation and odor generation of biomass and application thereof.

The technical scheme of the invention is as follows:

the invention provides an odor-inhibiting microorganism composition, which comprises one or more of yeast, mature bacteria or nitrobacteria; when the three are combined, the mass ratio of the saccharomycetes, the rotten bacteria and the nitrifying bacteria is (1-3) to (1-3).

The yeast is Saccharomyces cerevisiae NAU-OR3, is classified and named as Saccharomyces cerevisiae, and is preserved in Guangdong province microorganism culture collection center at 8-18 months in 2021, and has the address: no. 59 building 5 of No. 100 Dazhong Jie of Pieli, Guangzhou city, with the collection number GDMCC No. 61877.

The decomposing bacteria are Bacillus licheniformis NAU-OR5, are classified and named as Bacillus licheniformis, and are preserved in Guangdong province microorganism strain preservation center at 8-18 months in 2021, and the addresses are as follows: no. 59 building 5 of No. 100 Dazhong Jie of Pieli, Guangzhou city, with the collection number GDMCC No. 61879.

The nitrifying bacteria are Paracoccus denitificas NAU-OR1, are classified and named as Paracoccus denitificas, are stored in Guangdong province microorganism culture collection center at 10-28 months in 2021, and have the addresses: no. 59 building 5 of No. 100 college of Pieli Zhonglu, Guangzhou city, with the collection number GDMCC No. 61875).

In some embodiments, the mass ratio of the yeast, the mature bacteria and the nitrifying bacteria is (1-2): 1-3); in some more specific examples, the mass ratio of the yeast, the mature bacteria and the nitrifying bacteria is 1 (2-3) to (1-3).

The invention also provides a preparation method of the odor-inhibiting microorganism combination, which comprises the steps of mixing the strains in proportion; the preparation of each strain can be performed according to conventional methods in the art, and in one specific example comprises the following steps: respectively activating each strain stored at-80 ℃ on a plate culture medium, selecting a single colony to be cultured in a liquid culture medium to logarithmic phase, then transferring to a seed culture medium, and carrying out step-by-step enlarged culture; and (4) centrifuging and collecting thalli and spores. During the enlarged culture, components beneficial to the spore production of the thalli can be added.

The invention also provides an odor inhibiting microbial inoculum which comprises the odor inhibiting microbial composition and organic silicon, wherein the weight ratio of the odor inhibiting microbial composition to the organic silicon is (10-50): 1, preferably 10: 1.

the invention also provides the odor inhibiting microorganism combination or the application of the odor inhibiting microbial inoculum in treating biomass which generates malodor by fermentation.

The biomass capable of generating the malodor by fermentation can be any substance capable of generating gases such as ammonia gas, hydrogen sulfide, mercaptan, thioether, volatile organic compounds, comprehensive malodor and the like, and in a specific example, the biomass capable of generating the malodor by fermentation is livestock and poultry manure or kitchen garbage.

In one embodiment of the invention, when the kitchen waste is treated and inhibited, the mass ratio of the yeast, the rotten bacteria and the nitrifying bacteria is 1:2: 1.

In one embodiment of the invention, when the livestock manure is inhibited, the mass ratio of the yeast, the rotten bacteria and the nitrifying bacteria is 1:3: 2.

In some embodiments of the invention, when the odor-producing fermented biomass is kitchen waste, the odor-inhibiting microbial composition is present in an amount of greater than 3%, preferably from 3% to 10%, most preferably 3% by weight of the kitchen waste on a dry basis.

In some embodiments of the invention, when the odor-producing fermented biomass is livestock manure, the odor-inhibiting microbial composition is used in an amount of 6% or more, preferably 8 to 10% o, and most preferably 8% o, based on the dry weight of the livestock manure.

The invention also provides a using method of the odor-inhibiting microbial agent, the odor-inhibiting microbial composition and water are prepared into a bacterial liquid according to the ratio of 1:100, and organic silicon with the formula amount is added. The prepared odor-inhibiting microbial inoculum can be directly sprayed to biomass.

The invention provides an odor-inhibiting microbial composition, which is a novel biological odor inhibitor prepared by fermenting and compounding microorganism strains, wherein the effective components of saccharomycetes, decomposed bacteria and nitrobacteria are derived from kitchen garbage or livestock and poultry excrement. Because the screened microorganisms come from the production and living biomass waste, the biological safety of the microorganisms can be guaranteed; the odor inhibiting microbial inoculum can convert nutrient components in biomass, thereby inhibiting the generation of malodorous gases such as ammonia gas, hydrogen sulfide, methyl mercaptan, volatile organic compounds and the like from the source and well responding to the concept of environmental protection and emission reduction. The odor-inhibiting microorganism composition can be widely applied to places with odor generation, such as livestock farms, garbage transfer stations, garbage disposal plants, zoos, sewage treatment plants, sewage ponds (ditches), toilets, restaurants and the like.

In the invention, the odor inhibiting microorganism combination relates to a microorganism 'filtering' process when inhibiting the generation of odor from biomass, and the odor related substances are converted or the generated odor is degraded through the metabolic activity of microorganisms based on the absorption of metabolites and the fixed conversion of the microorganisms, and the specific process is as follows: (1) the decomposing microbial inoculum is used as a dominant strain to degrade macromolecules such as starch, cellulose, protein and the like in biomass into micromolecules; (2) part of the generated odor is dissolved in the odor inhibiting microbial inoculum and is converted into liquid phase from gas phase; (3) the yeast fixes and converts odor entering cells, and nitrobacteria can convert components which are easy to generate odor into nitrate or nitrite, so that the biomass is degraded and converted to inhibit the generation of odor. The conversion of the malodorous substances is the rate-limiting stage of the process, the microorganisms degrade the biomass and then serve as nutrients of other microorganisms to continue to absorb and digest, and the cycle is repeated to gradually convert and degrade the malodorous substances.

The use method of the odor-inhibiting microorganism combination provided by the invention can greatly improve the action effect, reduce the treatment cost to the maximum extent and is beneficial to large-area popularization and application.

The microbial combination of the invention has the capability of inhibiting the generation of comprehensive malodor of more than 99 percent; the contact angle of the bacterial liquid on the biomass can be obviously reduced by adding the organic silicon, the contact angle is reduced to 23.73 degrees from 53.75 degrees, the contact area of the microbial inoculum and the biomass is greatly increased, and the effect of inhibiting odor is further improved.

Drawings

FIG. 1 shows the odor components removed by 3 strains of microorganisms;

FIG. 2 shows the effect of different ratios on the odor of kitchen garbage;

FIG. 3 shows the effect of different ratios on the odor of the feces of livestock and poultry;

FIG. 4 shows the effect of different access amounts on the odor of kitchen waste;

FIG. 5 shows the effect of different dosages on the odor of the feces of livestock and poultry;

FIG. 6 shows the effect of different additive amounts of the adjuvant on the contact angle of the microbial inoculum.

Detailed Description

The combination of microorganisms and the method of using the same for efficiently inhibiting the generation of odor by fermentation of biomass according to the present invention will be described in detail with reference to the following examples, but they should not be construed as limiting the scope of the present invention. The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified.

The research of the invention finds that the concentration of ammonia gas after the kitchen garbage fermentation can be rapidly increased in a short period and reaches a peak value at 24 h; the comprehensive odor and the methyl mercaptan reach peak values in 72 hours, and both have a descending trend after reaching the peak values; comprehensive odor, hydrogen sulfide, methyl mercaptan and volatile organic compounds after the fermentation of the livestock and poultry manure reach peak values within 48 hours, and ammonia reaches peak values within 96 hours; the odor yield of the livestock and poultry manure is about 4 times of that of kitchen garbage. In the following examples, the two biomasses were selected for experiments, and it should be particularly noted that the application of the present invention is not limited to the two malodorous biomasses, and similar substances capable of generating gases such as ammonia, hydrogen sulfide, mercaptans, sulfides, volatile organic compounds, and synthetic malodors can achieve similar effects.

Unless otherwise stated, the pig manure used in the following examples was taken from a Satsuwa-day modern farm with initial yields of ammonia, hydrogen sulphide, methyl mercaptan, volatile organics and overall malodour of 18.08ppm/g, 2.19ppm/g, 0.29ppm/g, 1.26ppm/g dry manure and 690.15/g dry manure (dimensionless, in terms of per gram of dry manure, the same applies hereinafter). The initial yields of ammonia gas, hydrogen sulfide, methyl mercaptan, volatile organic compounds and comprehensive malodors in kitchen waste from a Pukou school canteen of Nanjing agriculture university are 3.66ppm/g, 0.03ppm/g, 1.98ppm/g, 1.72ppm/g dry waste and 97.10/g dry waste respectively.

The media used in the following examples are, unless otherwise specified, as follows:

the liquid culture medium and the seed culture medium of the yeast comprise the following components in percentage by weight: 10g/L of yeast extract, 20g/L of bactopeptone, 20g/L of agar, 200g/L of potato and 20g/L of glucose;

the liquid culture medium and the seed culture medium of the thoroughly decomposed bacteria are proportioned as follows: 10g/L of tryptone, 5g/L of yeast extract, 5g/L of sodium chloride and 1mL/L of 1N sodium hydroxide;

the liquid culture medium and the seed culture medium of the nitrifying bacteria are proportioned as follows: 5g/L of ammonium sulfate, 0.5g/L of magnesium sulfate, 0.7g/L of monopotassium phosphate and 0.5g/L of calcium chloride.

EXAMPLE 1 isolation, identification and preservation of three strains

The yeast is separated from kitchen garbage, 10.0g (wet weight) of the kitchen garbage is added into 20mL of water and stirred uniformly; slightly precipitating, taking 100 mu L of supernatant, spreading the supernatant on YPD medium added with 100mg/mL penicillin and levofloxacin, after growing out, selecting oval-shaped colony for purification culture, and sending a functional strain to sample and sequencing through functional verification, wherein the ITS sequence is shown as SEQ ID NO. 1. After comparison, the strain is determined to be Saccharomyces cerevisiae (Saccharomyces cerevisiae) NAU-OR3, is classified and named as Saccharomyces cerevisiae, is preserved in Guangdong province microorganism strain preservation center at 18 months 8 in 2021, and has the preservation number as follows: GDMCC 61877.

The decomposed bacteria are separated from the primarily fermented livestock and poultry manure, 10.0g (wet weight) of the primarily fermented livestock and poultry manure is added into 20mL of water and stirred uniformly; slightly precipitating, taking 100 mu L of supernatant, coating the supernatant on an LB culture medium plate, after the supernatant grows out, selecting a single colony for purification culture, and sequencing a strain with functions after function verification, wherein the 16sRNA sequence is shown as SEQ ID NO. 2. The Bacillus licheniformis (Bacillus licheniformis) NAU-OR5 is determined by comparison, is classified and named as Bacillus licheniformis, and is preserved in Guangdong province microorganism strain preservation center at 8-18 months in 2021 with the preservation number as follows: GDMCC 61879.

The nitrifying bacteria are separated from the primarily fermented livestock and poultry manure, 10.0g (wet weight) of the primarily fermented livestock and poultry manure is added into 20mL of water and stirred uniformly; slightly precipitating, taking 100 mu L of supernatant, coating the supernatant on a nitrobacteria culture medium plate, after the supernatant grows out, selecting a single colony for purification culture, and sequencing a strain with functions through function verification, wherein the 16sRNA sequence is shown as SEQ ID NO. 3. After comparison, the strain is determined to be Paracoccus denitrificans (Paracoccus denitirica) NAU-OR1, which is classified and named as Paracoccus denitirica, and is preserved in Guangdong provincial microorganism strain collection center at 28 months 10 in 2021 with the preservation number as: GDMCC 61875.

EXAMPLE 2 preparation of the combination of odor-inhibiting microorganisms

The preparation method of each strain comprises the following steps: activating the strain preserved at the temperature of minus 80 ℃ on a plate culture medium, picking a single colony to a liquid culture medium for culturing to a logarithmic phase, then transferring the single colony to a seed culture medium, and carrying out step-by-step enlarged culture; centrifugally collecting thalli and spores; and adding components beneficial to spore production of thalli in the amplification culture.

According to the method, after the strains of the yeast, the mature bacteria and the nitrobacteria are respectively activated on a solid culture medium, selecting a single colony to be inoculated into 2mL of corresponding liquid culture medium, culturing to a logarithmic phase, then transferring into a seed culture medium (the ratio is 1:1000), and carrying out step-by-step enlarged culture; and (4) centrifuging and collecting thalli and spores. The activated solid culture medium of the yeast is as follows: 10g/L of yeast extract, 20g/L of bactopeptone, 20g/L of agar, 200g/L of potato, 20g/L of glucose and 18g/L of agar; the activated solid culture medium of the mature bacteria is as follows: 10g/L of tryptone, 5g/L of yeast extract, 5g/L of sodium chloride, 1mL/L of 1N sodium hydroxide and 15g/L of agar; the activated solid culture medium of the nitrifying bacteria is as follows: 5g/L of ammonium sulfate, 0.5g/L of magnesium sulfate, 0.7g/L of monopotassium phosphate, 0.5g/L of calcium chloride and 18g/L of agar; when the microbial inoculum is produced in a large tank, the carbon source and the nitrogen source are required to be adjusted into starch, soybean meal, amylase and the like (the adjustment can be carried out according to a conventional method in the field), and a defoaming agent is added; in order to promote the spore formation of the bacterial cells, a certain amount of molasses is added (the amount may be adjusted according to a conventional method in the art). The culture temperature of the strain is 30-35 ℃. And centrifuging and collecting thalli and spores to obtain the corresponding bacterial strain.

The odor-inhibiting microorganism combination can be directly prepared into bacterial liquid with water according to the ratio of 1:100 and sprayed onto biomass.

The embodiment also provides a preparation and use method of the odor-inhibiting microbial agent, which comprises the steps of preparing a bacterial liquid from the prepared odor-inhibiting microbial composition and water according to a ratio of 1:100, adding organic silicon, and spraying the bacterial liquid on biomass.

In the invention, under the condition of not spraying any microbial inoculum, odor components and yields of different biomass materials are different, wherein the yields of ammonia gas, hydrogen sulfide, methyl mercaptan, volatile organic compounds and comprehensive odor of pig manure are respectively 18.08ppm/g, 2.19ppm/g, 0.29ppm/g, 1.26ppm/g dry manure and 690.15/g dry manure (dimensionless, in terms of per gram of dry manure, the same applies hereinafter); the saccharomyces cerevisiae sprayed independently has strong capability of fixing and converting nitrogen, and the yield of ammonia is only about 5%; the capacities of the bacillus licheniformis for inhibiting methyl mercaptan, hydrogen sulfide, comprehensive malodor and ammonia gas are respectively 100%, 99.29%, 98.93% and 85.81%, but the capacity for inhibiting the generation of volatile organic compounds is weaker; the capability of the nitrobacteria for inhibiting the generation of odor is relatively uniform, and the capabilities of inhibiting ammonia gas, hydrogen sulfide, methyl mercaptan, volatile organic compounds and comprehensive malodor are respectively 80.25%, 82.80%, 79.02%, 30.43% and 82.97%; when 3 strains of bacteria are compounded according to the ratio of 1:2:1 or 1:3:2, the deodorization capability is higher than the simple superposition of the three strains of bacteria, so that the selected combined microorganisms have the functions of complementation and synergism and have extremely strong capability of inhibiting the decomposition and odor generation of biomass.

EXAMPLE 3 odor inhibiting Effect of the combination of microorganisms

1. Materials and methods

The equipment used is shown in Table 1.

TABLE 1 instruments and apparatus

Chemical reagents: potassium sulfate and glucose are purchased from chemical reagents of national medicine group, ltd, and are analytically pure; copper sulfate was analytically pure and purchased from Aladdin reagents, Inc. Pig manure is taken from the Rugaohua-Ri modern farm and kitchen garbage is taken from a Pukou school canteen of Nanjing agriculture university.

2. Test method

2.1 Collection of somatic cells

Activating according to the method of the embodiment 2, picking single colony of the activated yeast, the activated mature bacteria and the activated nitrobacteria, inoculating the single colony into 2mL of corresponding liquid culture medium, placing the liquid culture medium in an incubator at 30 ℃, and shaking and drum the liquid culture medium for 120r/min overnight culture; and (3) inoculating 300 mu L of culture solution into 300mL of liquid culture medium, culturing at 160r/min until logarithmic phase, wherein 6 hours are required for yeast and mature bacteria, and 24 hours are required for nitrobacteria. Centrifuging at 10000r/min for 10min to collect thallus.

2.2 measurement of odor-suppressing Effect

The bacteria collected in 2.1 and water are prepared into bacteria liquid according to the ratio of 1:100, and the capabilities of 3 bacteria agents in inhibiting ammonia gas, hydrogen sulfide, mercaptan, thioether, volatile organic compounds and comprehensive odor are tested by taking water as a reference. Formula for calculating inhibition ratio (Y): y ═ Y water-Y microbial inoculum)/Y water x 100%

2.3 combined effect experiment of microorganisms

Yeast, decomposed bacteria and nitrobacteria collected at 2.1 are prepared into bacterial liquid according to the mass ratio of 3:1:1, 2:2:1, 1:1:1, 1:2:1, 1:3:2 and 1:3:3 of the bacteria, the bacterial liquid is respectively sprayed onto 5kg of kitchen garbage or livestock and poultry manure in the amount of 3 per thousand and 8 per thousand, and the reduction condition of various odor indexes is measured after 5 hours.

2.4 Effect experiment of inoculum size of Oesophagostimulant

Weighing odor-inhibiting microbial inoculum by 1 per thousand, 2 per thousand, 3 per thousand, 6 per thousand, 8 per thousand and 10 per thousand of dry biomass, preparing bacterial liquid according to the ratio of 1:100, spraying the bacterial liquid on 5kg of kitchen garbage or livestock and poultry manure, and measuring the reduction condition of various odor indexes after 5 hours.

2.5 Effect test of auxiliary

Selecting triton, pure ester and organic silicon as auxiliary agents, and testing survival and contact angle change conditions of yeast, decomposed bacteria and nitrobacteria after 1 ‰ of addition of the auxiliary agents; and adding auxiliary agents by 0.5 per mill, 1 per mill, 3 per mill and 5 per mill of the bacterial liquid, and measuring the change condition of the contact angle.

2.6 data processing and analysis

The obtained data were statistically analyzed using IBM SPSS Statistics 25, Excel 2016, Origin 2018 software; the significance difference between the different treatments was examined using Duncan, α ═ 0.05.

3 results of the experiment

3.1 odor inhibiting Effect of the three microorganisms of the present invention

4 microbial strains are selected in research, and a chemical deodorization reagent (provided by university of Hospital) and deionized water are used as a control to compare the odor inhibiting effect, as shown in figure 1, the results show that the efficiency of the bacillus licheniformis for inhibiting ammonia gas, hydrogen sulfide, methyl mercaptan and comprehensive malodor reaches 85.81%, 99.29%, 100.00% and 98.93% respectively, wherein the inhibition capacity for hydrogen sulfide and comprehensive malodor is obviously higher than that of other treatments (P < 0.05); the generation of volatile organic compounds (eVOC) cannot be greatly inhibited in each treatment, but the nitrobacteria and the yeast respectively reach 30.43% and 27.70%, which are obviously higher than those in other treatments (P <0.05), and the nitrobacteria, the yeast and the rotten bacteria have good odor removing effect.

3.2 odor inhibiting effect of different proportions of microorganisms

Yeast, mature bacteria and nitrobacteria are prepared into bacteria liquid according to the mass ratio of bacteria of 3:1:1, 2:2:1, 1:1:1, 1:2:1, 1:3:2 and 1:3:3, and the bacteria liquid is respectively sprayed on kitchen garbage and livestock and poultry excrement in the amount of 3 per thousand and 8 per thousand, and as a result, the result shows that the prepared odor-inhibiting microbial combination of the mature bacteria, the nitrobacteria and the yeast can make up the defects of each strain in the aspect of inhibiting the generation of volatile organic matters, can exert the advantages of bacillus licheniformis in inhibiting ammonia gas, hydrogen sulfide, mercaptan, thioether and comprehensive odor, has a remarkable deodorization effect, has a remarkable odor-inhibiting effect in the range of the mixture ratio, has a better odor-inhibiting effect in the preferable range of the invention, and particularly when the ratio is 1:2:1, the ammonia gas produced by fermentation of the kitchen garbage, the hydrogen sulfide and the comprehensive odor are inhibited, The hydrogen sulfide and the comprehensive odor are obviously inhibited (P is less than 0.05), and the odor concentration is reduced by more than 90 percent compared with that of the odor without adding a microbial inoculum; the ratio of the components is 1:3:2, so that the generation of livestock and poultry excrement stink can be obviously inhibited (figure 2 and figure 3).

3.3 inhibition of odor production by fermentation of Biomass with different microbial inoculum sizes

The odor inhibiting microbial inoculum is weighed by 1 per thousand, 2 per thousand, 3 per thousand, 6 per thousand, 8 per thousand and 10 per thousand of the dry basis of the kitchen waste and the livestock and poultry manure, and is prepared into a bacterial solution according to the ratio of 1:100 and then sprayed on the kitchen waste and the livestock and poultry manure, and tests show that within the range of the inoculation amount of the microbial inoculum provided by the invention, the odor inhibiting effect with an obvious effect can be realized, particularly, the effect which is 3 per thousand and more than the kitchen waste is obviously better than that of 1 per thousand and 2 per thousand treatment (P <0.05) when the inoculation amount of the microbial inoculum reaches 6 per thousand and more than that of the livestock and poultry manure (P >0.05), and from the economical point of view, the 3 per thousand and 8 per thousand are respectively the optimal inoculation amount of the kitchen waste and the livestock and poultry manure (figures 4 and 5).

3.4 Effect of the auxiliary on the combination of odor-inhibiting microorganisms

After the microbial composition is added with assistants such as triton, pure ester or organic silicon and the like, the number of bacteria is not obviously reduced (P is more than 0.05) compared with that of bacteria added with sterile water through plate counting; further comparing the contact angle changes of different additives and no additive, the contact angle is reduced from 53.75 degrees to 23.73 degrees only after the organic silicon is added, and the infiltration of the microbial inoculum on the surface of the garbage is increased; in addition, the addition amount of the additives such as 0.5%, 1%, 3%, 5% and the like can greatly reduce the contact angle, and the addition amount of the additives of 1% or more is optimal from the economical point of view because the effect is increased and relaxed by 1% or more (fig. 6).

Example 4 inhibition of sheep manure odor

According to the reflection of the Satsugao Huari modern farm, sheep manure generated by the Satsugao Huari modern farm is difficult to treat and has a large odor; through detection, the comprehensive odor reaches 658.78/g dry feces; for the microbial combination prepared under the conditions of the farm manure example 2, namely, after the yeast, the thoroughly decomposed bacteria and the nitrobacteria are compounded according to the ratio of 1:3:2, 1 thousandth of the organic silicon additive is added, and spraying is carried out according to 8 thousandth of the dry weight of the sheep manure, so that after 5 hours, the comprehensive odor index is reduced by 92.6 percent compared with the initial value.

Example 5 suppression of odor of canteen leftovers

In this test, 2kg of the meal left in the canteen was placed in three plastic bottles (numbered #1, #2, and #3) of the same volume, respectively, and the bottle mouth was closed to allow the bottles to ferment at 35 ℃ and accumulate odor.

Opening two bottles #1 and #2 after half a month, wherein both bottles have foul smell; 3 per mill (by initial dry weight) of the microbial composition using the conditions described in example 2 was sprayed into bottle 1, i.e. 1 per mill of the silicone adjuvant was added after yeast, decomposing bacteria and nitrifying bacteria were compounded at 1:2: 1; at the subsequent 24 hours, the odor of both bottles changed: the odor in the bottle No.1 sprayed with the odor inhibitor is obviously lightened, and the odor in the bottle No.2 is not obviously changed; the bottles #1 and #2 were sealed and left to stand.

After one month, three bottles were opened and the #1, #2, #3 odors were compared. Odor indexes such as ammonia gas, hydrogen sulfide, mercaptan, thioether, volatile organic compounds, comprehensive odor and the like in the bottle #1 are obviously lower than those of the bottles #2 and #3, and no obvious difference exists between the odor indexes of the bottles #2 and # 3. Through the experiment, the microbial composition is verified to have an obvious inhibiting effect on the generation of odor, and the effect is obvious.

The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.

Sequence listing

<110> Nanjing university of agriculture

<120> microbial composition for efficiently inhibiting odor production by biomass fermentation at source and application thereof

<160> 3

<170> SIPOSequenceListing 1.0

<210> 1

<211> 727

<212> DNA

<213> Saccharomyces cerevisiae

<400> 1

tttttttgtt ttggcaagag catgagagct tttactgggc aagaagacaa gagatggaga 60

gtccagccgg gcctgcgctt aagtgcgcgg tcttgctagg cttgtaagtt tctttcttgc 120

tattccaaac ggtgagagat ttctgtgctt ttgttatagg acaattaaaa ccgtttcaat 180

acaacacact gtggagtttt catatctttg caactttttc tttgggcatt cgagcaatcg 240

gggcccagag gtaacaaaca caaacaattt tatctattca ttaaattttt gtcaaaaaca 300

agaattttcg taactggaaa ttttaaaata ttaaaaactt tcaacaacgg atctcttggt 360

tctcgcatcg atgaagaacg cagcgaaatg cgatacgtaa tgtgaattgc agaattccgt 420

gaatcatcga atctttgaac gcacattgcg ccccttggta ttccaggggg catgcctgtt 480

tgagcgtcat ttccttctca aacattctgt ttggtagtga gtgatactct ttggagttaa 540

cttgaaattg ctggcctttt cattggatgt tttttttcca aagagaggtt tctctgcgtg 600

cttgaggtat aatgcaagta cggtcgtttt aggttttacc aactgcggct aatctttttt 660

tatactgagc gtattggaac gttatcgata agaagagagc gtctaggcga acaatgttct 720

taaagtt 727

<210> 2

<211> 1374

<212> DNA

<213> Bacillus licheniformis (Bacillus licheniformis)

<400> 2

tcgagcggac cgacgggagc ttgctccctt aggtcagcgg cggacgggtg agtaacacgt 60

gggtaacctg cctgtaagac tgggataact ccgggaaacc ggggctaata ccggatgctt 120

gattgaaccg catggttcaa ttataaaagg tggcttttag ctaccactta cagatggacc 180

cgcggcgcat tagctagttg gtgaggtaac ggctcaccaa ggcgacgatg cgtagccgac 240

ctgagagggt gatcggccac actgggactg agacacggcc cagactccta cgggaggcag 300

cagtagggaa tcttccgcaa tggacgaaag tctgacggag caacgccgcg tgagtgatga 360

aggttttcgg atcgtaaaac tctgttgtta gggaagaaca agtaccgttc gaatagggcg 420

gtaccttgac ggtacctaac cagaaagcca cggctaacta cgtgccagca gccgcggtaa 480

tacgtaggtg gcaagcgttg tccggaatta ttgggcgtaa agcgcgcgca ggcggtttct 540

taagtctgat gtgaaagccc ccggctcacc cggggagggt cattggaaac tggggaactt 600

gagtgcagaa gaggagagtg gaattccacg tgtagcggtg aaatgcgtag agatgtggag 660

gaacaccagt ggcgaaggcg actctctggt ctgtaactga cgctgaggcg cgaaagcgtg 720

gggagcgaac aggattagat accctggtag tccacgccgt aaacgatgag tgctaagtgt 780

tagagggttt ccgcccttta gtgctgcagc aaacgcatta agcactccgc ctggggagta 840

cggtcgcaag actgaaactc aaaggaattg acgggggccc gcacaagcgg tggagcatgt 900

ggtttaattc gaagcaacgc gaagaacctt accaggtctt gacatcctct gacaacccta 960

gagatagggc ttccccttcg ggggcagagt gacaggtggt gcatggttgt cgtcagctcg 1020

tgtcgtgaga tgttgggtta agtcccgcaa cgagcgcaac ccttgatctt agttgccagc 1080

attcagttgg gcactctaag gtgactgccg gtgacaaacc ggaggaaggt ggggatgacg 1140

tcaaatcatc atgcccctta tgacctgggc tacacacgtg ctacaatggg cagaacaaag 1200

ggcagcgaag ccgcgaggct aagccaatcc cacaaatctg ttctcagttc ggatcgcagt 1260

ctgcaactcg actgcgtgaa gctggaatcg ctagtaatcg cggatcagca tgccgcggtg 1320

aatacgttcc cgggccttgt acacaccgcc cgtcacacca cgagagtttg taac 1374

<210> 3

<211> 1285

<212> DNA

<213> Paracoccus denitrificans (Paracoccus denitiricans)

<400> 3

aacccttggg ggttagcggc ggacgggtga gtaacgcgtg ggaatatgcc ctttgctacg 60

gaatagcccc gggagactgg gattaatacc gtatacgccc tttgggggag agatttatcg 120

gcaaaggatt agcccgcgtt ggattaggta gttggcgggg taatggccta ccaagccgac 180

gatccatagc tggtttgaga ggatgatcac ccacactggg actgagacac ggcccagact 240

cctacgggag gcagtggggg ggaatcttag acaatggggg caaccctgat ctagccatgc 300

cgcgtgagtg atgaaggccc tagggttgta aagctctttc agctgggaag ataatgacgg 360

taccagcaaa agaagccccg gctaactccg tgccagcagc cgcggtaata cggagggggc 420

tagcgttgtt cggaattact gggcgtaaag cgcacgtagg cggaccggaa agttgggggt 480

gaaatcccgg ggctcaacct cggaactgcc ttcaaaacta tcggtctgga gttcgagaga 540

ggtgagtgga attccgagtg tagaggtgaa attcgtagat attcggagga acaccagtgg 600

cgaaggcggc tcactggctc gatactgacg ctgaggtgcg aaagcgtggg gagcaaacag 660

gattagatac cctggtagtc cacgccgtaa acgatgaatg ccagtcgtcg ggcagcatgc 720

cgttcggtga cacacctaac ggattaagca ttccgcctgg ggagtacggt cgcaagatta 780

aaactcaaag gaattggcgg ggggccgcac aagcggtgga gcatgtggtt taattcgaag 840

ctacgcgcag aaccttacca tcccttgaca tcgcaggccc gctccagaga tggagttttg 900

ttgtaagagg cctgtggtca ggtggtgcat ggctgtcgtc agctcgtgtc gtgagatgtt 960

cggttaagtc ctgcaacgag cgcacccccc actgttagtt gccagcattt ggttgggcac 1020

tataagagaa ctgccgatga taagtcggag gaaggtgtgg atgacgtcaa gtcctcatgg 1080

cccttacggg ttgggctaca cacgtgctac aatggtggtg acagtgggtt aatccccaaa 1140

agccatctca gttcggattg gggtctgcaa ctcgacccca tgaagttgga atcgctagta 1200

atcgcggaac agcatgccgc ggtgaatacg ttcccgggcc ctgtacacac cccccgtcac 1260

accacgggag ttgggtctac cccac 1285

Claims

1. An odor-inhibiting microbial composition comprising one or more of yeast, spoilage bacteria, or nitrifying bacteria; when the three are combined, the mass ratio of the saccharomycetes, the rotten bacteria and the nitrifying bacteria is (1-3) to (1-3) respectively; the yeast is Saccharomyces cerevisiae NAU-OR3 with the preservation number of GDMCC No. 61877, the rotten bacteria is Bacillus licheniformis NAU-OR5 with the preservation number of GDMCC No. 61879), the nitrifying bacteria is Paracoccus densificans NAU-OR1 with the preservation number of GDMCC No. 61875.

2. The odor-inhibiting microbial composition according to claim 1, wherein the mass ratio of the yeast, the decomposing bacteria and the nitrifying bacteria is (1-2) to (1-3); preferably, the mass ratio of the yeast, the rotten bacteria and the nitrifying bacteria is 1 (2-3) to (1-3).

3. The method for producing an odor-inhibiting microbial composition according to claim 1 or 2, wherein a proportional amount of each of the strains is mixed.

4. An odor-inhibiting microbial inoculum, which comprises the odor-inhibiting microbial composition as claimed in claim 1 or 2 and organic silicon, wherein the weight ratio of the odor-inhibiting microbial composition to the organic silicon is (10-50): 1, preferably 10: 1.

5. use of a combination of odour-inhibiting microorganisms according to claim 1 or 2 or of an odour-inhibiting bacterial agent according to claim 3 for the treatment of a biomass which is fermented to produce malodours.

6. The use according to claim 5, wherein the fermented malodorous biomass is any substance capable of producing one or more of ammonia gas, hydrogen sulfide, mercaptans, thioethers, volatile organics, or complex malodorous gases; preferably, the biomass capable of generating the malodor by fermentation is livestock and poultry manure or kitchen waste.

7. The use according to claim 5, wherein the mass ratio of the yeast, the decomposing bacteria and the nitrifying bacteria is 1:2:1 when treating the kitchen waste.

8. The use according to claim 5, wherein the mass ratio of the yeast, the rotting fungi and the nitrifying bacteria is 1:3:2 when the livestock manure is treated.

9. Use according to claim 5, wherein, when the fermented malodorous biomass is kitchen waste, the combination of odour-inhibiting microorganisms is used in an amount of more than 3%, preferably 3-10%, most preferably 3% by weight of the kitchen waste on a dry basis.

10. Use according to claim 5, wherein, when the fermented malodorous biomass is livestock manure, the amount of the combination of odor-inhibiting microorganisms is more than 6%, preferably 8-10%, most preferably 8% of the dry weight of the livestock manure.