TWI766554B - Microbial deodorizing reagent, method for deodorizing compost and use of mixed strains - Google Patents

Abstract

The present invention provides a microbial deodorizing reagent. The microbial deodorizing reagent includes mixed strains. The mixed strains include a first group of strains. The first group of strains is at least one strain selected from the group consisting of Rhodococcussp. and Acinetobacter towneri.The Rhodococcussp. is given accession number of BCRC 911001 and deposited in Food Industry Research and Development Institute. The Acinetobacter towneri.is given accession number of BCRC 911002 and deposited in Food Industry Research and Development Institute.

Description

Microbial deodorant preparation, deodorization method of compost and use of mixed strains

The present invention relates to a deodorant preparation and a compost deodorization method, in particular to a composite microbial deodorant preparation, a compost deodorization method using the aforementioned microbial deodorant preparation, and the use of mixed strains.

Composting is a mixture of organic materials such as biological residues, livestock manure, substrates, etc., to an appropriate carbon-nitrogen ratio and moisture content, and is transformed, fermented and decomposed into materials that can be used by plants. In addition, the high temperature generated by fermentation during the composting process can be used to kill pathogenic bacteria, insect eggs and weed seeds in the composting source, thereby achieving the purpose of harmlessness.

During the composting process, it is necessary to add organic sources with high nitrogen content such as livestock manure, and adjust the carbon-nitrogen ratio to an appropriate range to promote microbial reproduction and accelerate the maturation of the compost. However, the addition of livestock manure can also lead to an increase in foul odors during composting. The volatilization of ammonia is the main cause of malodorous odor and nitrogen loss during composting. Although ammonia is not the only odor molecule that contributes to the odor of livestock manure, the volatilization of ammonia is closely related to the volatilization of other malodorous odors in manure. Therefore, if the volatilization of ammonia and other odor molecules can be properly controlled, the emission of odor and nitrogen loss can be effectively reduced.

Based on the above, there is still a need for an effective microbial deodorant preparation to reduce the odor generated during the composting process, so as to improve the environment of the composting field, and at the same time, it can also improve the composting efficiency and quality, and recycle the natural resources, so as to solve the problem of environmental protection question.

The present invention provides a microbial deodorant preparation, which comprises a mixed strain including a first group of strains, wherein the first group of strains is selected from Rhodococcus sp. At least one of Acinetobacter towneri ), and wherein the Rhodococcus is deposited in the Food Industry Development Research Institute of the Republic of China Consortium under the deposit number BCRC 911001, and the Acinetobacter thomii is deposited in the Food Industry Development Research Institute of the Republic of China Consortium So, deposit number BCRC 911002.

In addition, the mixed strains in the above-mentioned microbial deodorant preparation may further include a second group of strains, and the second group of strains are selected from Comamonas testosteroni , Alcaligenes faecalis and Contaminate at least one of Brevibacillus invocatus .

The present invention also provides a method for deodorizing compost, comprising the following steps: (a) mixing a biological waste with a substrate to form compost with a carbon-nitrogen ratio between 30 and 40; compost applying the microbial deodorizing formulation described above to form a compost mixture; (c) subjecting the compost mixture to a first turn; and (d) allowing the compost mixture to stand for up to 14 days to allow the hybrid strain to compost the compost Fermentation and decomposition of odor molecules produced by the biological waste.

The present invention also provides the use of a mixed strain for compost deodorization, wherein the mixed strain includes a first group of strains, wherein the first group of strains is selected from at least one of Rhodococcus and Acinetobacter tonneii , and wherein the Rhodococcus system is deposited in the Food Industry Development Research Institute of the Republic of China Consortium, with the deposit number BCRC 911001, and the Acinetobacter tangmani is deposited in the Food Industry Development Research Institute of the Republic of China Consortium, and the deposit number is BCRC 911002.

The present invention also provides the use of a mixed strain for compost deodorization, wherein the mixed strain is composed of Comamonas testosterone, Rhodococcus, Alcaligenes faecalis, Acinetobacter tanninii and Bacillus contaminus.

The present invention can provide a microbial deodorant preparation. In the process of compost maturation, with the frequency of turning the compost at a specific interval and applying the microbial deodorant preparation again, the compost can maintain a proper temperature and moisture content, which is conducive to the fermentation process of the compost, and also makes the mixed strain in the microbial deodorant preparation more efficient. Efficiently degrade odor molecules, thereby achieving the purpose of reducing environmental pollution. The microbial preparation of the present invention can achieve deodorization effect by degrading odor molecules, and can be suitable for deodorization of compost. The odor molecules mentioned herein refer to any odor molecules that can be perceived by the human sense of smell, especially odor molecules that cause subjective unpleasantness. For example, odor molecules may be odor molecules produced by biological waste. Examples of odor molecules may include, but are not limited to, ammonia, methylamine, phenol, cresol, indole, derivatives of the foregoing, or combinations of the foregoing. Examples of the odor molecules listed above are the volatile odor molecules that mainly contribute to odor in biological excrement, and can also be the odor molecules described in the present invention.

In one embodiment, the example of the above-mentioned microbial deodorant formulation includes a mixed strain. The aforementioned mixed strains may include the first group of strains. The above-mentioned first group of strains may include at least one of Rhodococcus and Acinetobacter Thomsonii.

In a specific embodiment, the mixed strains in the above-mentioned microbial deodorant preparation may further include a second group of strains, and the second group of strains may include any of Comamonas testosterone, Alcaligenes faecalis, and Brevibacterium contamination. at least one of them.

Examples of the above-mentioned C. testosterone may include C. testosterone deposited in the Food Industry Development Research Institute of the Republic of China with the deposit number of BCRC 911003, but not limited thereto.

Examples of the above-mentioned Rhodococcus may include, but not limited to, Rhodococcus deposited with the Food Industry Development Research Institute of the Republic of China with the deposit number BCRC 911001.

Examples of the above-mentioned Alcaligenes faecalis include, but not limited to, Alcaligenes faecalis deposited with the Food Industry Development Research Institute of the Republic of China with the deposit number of BCRC 911000.

Examples of the above-mentioned Acinetobacter tumefaciens may include, but are not limited to, the deposit number BCRC 911002 at the Food Industry Development Research Institute of the Republic of China.

Examples of the above-mentioned contaminating Brevibacterium may include, but are not limited to, the deposit number BCRC 911004 at the Food Industry Development Research Institute of the Republic of China.

In a specific embodiment, the microbial deodorant formulation may include a combination of Comamonas testosterone, Rhodococcus, Ralcaligenes faecalis, Acinetobacter tannii, and Brevibacterium contagiosum, wherein Comamonas testosterone It can be deposited with the Food Industry Development Research Institute of the Republic of China Consortium, with the deposit number BCRC 911003, and Rhodococcus can be deposited with the Food Industry Development Institute of the Republic of China, with the deposit number BCRC 911001 Rhodococcus faecalis, Alcaligenes faecalis can be deposited in the Food Industry Development Research Institute of the Republic of China Consortium, and the deposit number is BCRC 911000. The deposit number is BCRC 911002, and the contaminating Brevibacillus can be deposited at the Food Industry Development Research Institute of the Republic of China Foundation, and the deposit number is BCRC 911004.

The bacterial ratio of Comamonas testis ketone, Rhodococcus, Alcaligenes faecalis, Acinetobacter tangneri and Bacillus contaminating Brevibacterium can be about 1:0.5-1.5:0.5-1.5:0.5-1.5:0.5- 1.5, for example, may be about 1:0.6-1.4:0.6-1.4:0.6-1.4:0.6-1.4, or may be about 1:0.7-1.3:0.7-1.3:0.7-1.3:0.7-1.3, or may be about 1:0.8-1.2:0.8-1.2:0.8-1.2:0.8-1.2, or approximately 1:0.9-1.1:0.9-1.1:0.9-1.1:0.9-1.1, or approximately 1:1:1: 1:1, but not limited to this.

The microbial deodorant formulation can be in the form of, but not limited to, a suspension or a powder, and the like. The microbial deodorant preparation in the form of suspension may include the above-mentioned mixed strains and a solution for suspending the above-mentioned mixed strains, and the solution may include, but is not limited to, a tryptic soy broth (TSB). Medium components, for example, may include casein, soybeen, D-glucose, sodium chloride (NaCl), dipotassium hydrogen phosphate (K ₂ HPO ₄ ), etc., or a combination of the foregoing. Examples of the above solution may include water or TSB medium, etc., but are not limited thereto. The microbial deodorizing preparation in powder form may include the above-mentioned mixed strains and dry powder coagulation aids such as bran, soybean meal, soybean meal, and dextrin.

In a specific embodiment, the microbial deodorant preparation may be in the form of a suspension, and the above-mentioned total of Comamonas testosterone, Rhodococcus, R. The bacterial count can range from above about 10 ⁸ cfu/ml to about 10 ¹³ cfu/ml.

In addition, the present invention can also provide a method for deodorizing compost, which can achieve the effect of effectively deodorizing compost by using any of the above-mentioned microbial deodorizing preparations of the present invention. In the process of compost maturation, with the frequency of turning over at a specific interval and applying the microbial deodorizing agent again, the compost can maintain a proper temperature and moisture content, so as to promote the fermentation process of the mixed strains in the microbial deodorizing agent, and also make the microbial deodorization agent. The mixed strains in the odor preparation can degrade odor molecules more efficiently, thereby achieving the purpose of reducing environmental pollution.

The above-mentioned deodorizing method of compost includes the following steps. First, the biological waste is mixed with the substrate to form compost. The term "biological excrement" used herein may refer to the organic excrement produced by any animal, and the animal may be human, chicken, pig, cow or sheep and other organisms, but is not limited thereto. In addition, examples of the above-mentioned substrates may include wood chips, rice husks, straw, bark, fallen leaves, soybean meal, mushroom feet, bagasse, vegetable bagasse and peanut shells or combinations thereof, but the present invention is not limited thereto. The term "substrate" as used herein may refer to any agricultural biological waste produced by agricultural activities, and in particular to organic waste derived from the biomass of plants or fungi, such as wood chips, chaff, Straw, mushroom feet, bagasse, vegetable residue, soybean meal, branches, bark, grass clippings, flower waste, fallen leaves, wheat straw, reed, sunflower stalk, thatch stalk, rice straw, corn cob, corn stalk, peach pit, apricot pit, plum Nuts, peanut shells, sunflower seed shells, cottonseed shells, camellia nut shells, coconut bran, coconut shells, etc. or a combination of the foregoing. In embodiments of the present invention, the substrate may be used to provide the primary carbon source for the compost.

The carbon-nitrogen ratio and water content of the compost can be further adjusted by adjusting the ratio of biological waste to the substrate, so as to promote the reproduction of the mixed strains in the above-mentioned microbial deodorant preparation and accelerate the fermentation process of the compost. The carbon-nitrogen ratio of the above-mentioned compost may be between about 20 and about 50, for example, between about 30 and 40, but not limited thereto. The moisture content of the above-mentioned compost may be between about 40% and about 60%, for example, between about 45% and about 55%, but not limited thereto.

Next, the aforementioned microbial deodorizing formulation is applied to the compost to form a compost mixture. In one embodiment, the microbial deodorant formulations described herein may be in the form of a suspension. Also, in this embodiment, the microbial deodorizing formulation may be applied in an amount ranging from about 5 ml to about 30 ml, for example, between about 10 ml and about 25 ml, per kilogram of compost, or It can be between about 15ml to about 25ml. In a specific embodiment, the applied amount of the microbial deodorant formulation may be about 20 ml, but is not limited thereto.

Then, the above-mentioned compost mixture is turned for the first time and the compost mixture is left to stand, so that the mixed strains in the microbial deodorizing preparation ferment the compost and decompose the odor molecules produced by the biological waste. The standing days of the compost mixture may be up to 14 days, but the invention is not limited thereto. In other embodiments, the resting days of the compost mixture may be up to between about 6 days and about 14 days, eg, 8 days, 10 days, or 12 days. The maximum standing days of the compost mixture refers to the number of days in which the compost treated with the microbial deodorizer can significantly reduce the odor molecules produced by the biological waste compared to the case where the compost is not treated with the microbial deodorizer. In addition, it should be noted that in the process of standing the compost mixture, in addition to the steps of mixing the compost with the microbial deodorizing agent and turning the compost for the first time, it may also include the subsequent application of the microbial deodorizing agent to the compost mixture, and then again The step of turning the compost is carried out, and the further processing steps of the compost mixture will be described in detail below.

In one embodiment, in addition to the above-mentioned steps, the method for deodorizing compost may further include applying the same amount of microbial deodorizing agent to the compost mixture at an interval of frequency after turning the compost for the first time, and deodorizing the compost. The compost mixture is turned over. As the microbial strains ferment within the compost, a large amount of heat is generated. The high temperature of the compost will reduce the fermentative decomposition efficiency of microbial strains, and will also affect the growth of microbial strains. Therefore, in the process of compost deodorization, the compost may be turned again at a certain interval after the first turning, so as to reduce the overall temperature of the compost. At the same time, the same amount of microbial deodorant preparation as the previous one can be added to maintain the moisture content of the compost, and it can also supplement the mixed strains that have been reduced due to high temperature inactivation.

The above-mentioned interval can be between about 0.5 days and about 5.5 days, for example, it can be between about 1 day and about 5 days, or it can be between about 1.5 days and about 4.5 days, or it can be between about 2 days. Between days and about 4 days, or may be between about 2.5 days and about 3.5 days.

In addition to the above, the present invention can also provide the use of a mixed strain for compost deodorization, wherein the mixed strain includes the first group of strains discussed above. In particular, the first group of strains is selected from at least one of Rhodococcus and Acinetobacter tonneii.

Furthermore, the present invention can also provide the use of a mixed strain for compost deodorization, wherein the mixed strain can be composed of Comamonas testosterone, Rhodococcus, Ralcaligenes faecalis, Acinetobacter tonneii and Contaminated by Bacillus brevis.

Example

1. Strain screening and single strain deodorization test

First, about 190 microbial strains were isolated from the matured compost. The term "aged compost" here refers to compost that has been odorless after the fermentation process. The isolated microbial strains are individually cultured in sterilized chicken manure water or sterilized pig manure water, and strains that can grow stably in chicken manure water or pig manure water are selected. The stable growth of microbial strains in chicken manure water or pig manure water means that they can effectively utilize the nutrients contained in chicken manure water or pig manure water for fermentation to convert them into nutrient forms that can be used by themselves. Therefore, microbial strains that can grow stably in chicken manure water or pig manure water were selected for further deodorization tests described below.

Next, the selected single strains were individually cultured with 5ml of tryptic soy broth (TSB) medium to reach a stable stage, and the cells were collected by centrifugation, rinsed with reverse osmosis water and centrifuged twice at low speed. , and redissolved in 1ml of reverse osmosis water. Take 0.5ml of bacterial liquid and transfer it to 25ml of sterilized chicken manure water (ammonia nitrogen value of 380mL/L) or sterilized pig manure water (ammonia nitrogen value of 280mL/L) in a shake flask. After 5 days of shaking culture at 30 °C, headspace solid-phase microextraction combined with gas chromatography-mass spectrometry (HS-SPME/GC-MS) method was used to analyze chicken manure water or pig manure water treated with a single strain for 5 days. Volatile odor molecule content.

Specifically, 5 ml of the bacterial solution after 5 days of culture was transferred to a 20 mL sample bottle containing 1.5 g NaCl, shaken at 60°C and 500 rpm for 40 minutes, and then the CAR/PDMS 85 μm SPME fiber (Supelco , Bellefonte, PA, USA) was desorbed at 250°C for 10 minutes, inserted into a headspace vial for 40 minutes of adsorption, and then desorbed at the GC-MS injection port for 1 minute. Before each sample extraction and adsorption, the CAR/PDMS 85 μm SPME fiber must be desorbed at 250 °C for 10 minutes. Analysis was performed on an Agilent 7890B GC system with HP-5MS (30 m x 0.25 mm i.d. x 0.25 μm) coupled to an Agilent 5977 A mass spectrometer (Agilent Technologies, Palo Alto, CA, USA). Sampling temperature 250 DEG C; oven heating conditions are as follows: 40 DEG C is kept for 2 minutes, after warming up to 60 DEG C with 4 DEG C/min, keep 2 minutes, then be warming up to 190 DEG C at 2 DEG C/min, finally with The temperature was raised to 230°C at 5°C/min and kept for 15 minutes; the injection mode was splitless automatic injection; the carrier gas was helium, the flow rate was 1 mL/min, and the electron energy was 70 eV; the ion source temperature was 230°C; Transmission line temperature 280°C; quadrupole temperature 150°C; emission current 200 μA; scan 33-495 amu using full scan mode.

Then, using headspace solid-phase microextraction combined with the analysis results of gas chromatography-mass spectrometry, the odor molecules were compared in the NIST (National Institute of Standards and Technology) mass spectrometry database (NIST14.L), and the comparison was similar. The odor molecules with a property of more than 70% were accumulated, and the curve area integral of each odor molecule in the map was accumulated to calculate the total content of odor molecules after treatment with a single strain of each group. After comparing with the chicken manure water or pig manure water before treatment and the control group (reverse osmosis (RO) water only), 5 strains with significant potential for degrading odor molecules were screened out, and they were listed as follows. The 5 strains were further identified by 16S rDNA molecular as described above.

The above five strains were obtained from tryptone Soy Agar (TSA) culture plates after culturing for several days, collected in a 2 mL microcentrifuge tube, added with 0.5 mL of sterile water, and shaken to uniformly disperse the bacterial cells. , using the genomic DNA extraction kit ZR Fungal/Bacterial DNA MiniPrep ^™ (ZYMO RESEARCH) kit for DNA extraction. The bacterial genomic DNA was used as a PCR template, and its gene fragment was amplified with the relevant primer set in the 16S rRNA region (Winsley, et al ., 2012). The PCR reaction solution is as follows: Take an appropriate amount of genomic DNA solution as a PCR template, add 8 µL of 10 mM dNTP, 10 µL of 10X PCR buffer, 10 pmole of 5-terminal primer and 3-terminal primer and 5 U Taq enzyme. PCR reaction conditions were 95°C, 3 minutes; (95°C, 30 seconds, 50°C, 30 seconds, 72°C, 2 minutes, 30 seconds) for a total of 30 cycles; 72°C, 10 minutes; and finally kept at 4°C. The PCR products were purified and sequenced, and the sequence results were recombined and sequenced with Vector NTI Suite 9 software (VNTI) and NCBI/Blastn (http://www.ncbi.nlm.nih.gov/BLAST/) database. Similarity comparison analysis.

After 16S rDNA molecular identification, the five strains were confirmed to be Comamonas testosteroni , Rhodococcus sp., Alcaligenes faecalis , and Acinetobacter towneri . and the contaminating Brevibacillus invocatus . The 5 strains isolated by the present invention were all deposited in the Food Industry Development Research Institute of the Republic of China Consortium on May 26, 2020. Among them, Comamonas testosterone was named Comamonas testosterone C1-2/ new ( Comamonas testosteroni C1-2/new), whose deposit number is BCRC 911003; Rhodococcus sp. D2-3 ( Rhodococcus sp. D2-3), whose deposit number is BCRC 911001; Alcaligenes faecalis name It is Alcaligenes faecalis DO-BC3-1 ( Alcaligenes faecalis DO-BC3-1), its deposit number is BCRC 911000; Acinetobacter towneri is named as Acinetobacter towneri DO-Mix-7-1 ( Acinetobacter towneri DO-Mix -7-1), whose deposit number is BCRC 911002; and the contaminating Brevibacillus is named as Brevibacillus invocatus G08, whose deposit number is BCRC 911004.

The odor molecule degradation rate of above-mentioned 5 strains in chicken manure water or pig manure water, with the odor molecule content of the chicken manure water or pig manure water before treatment as 100% (that is, the degradation rate is 0%), They are shown in Table 1 below.

Table 1 - Degradation rate of odor molecules by a single deodorant strain in chicken manure water or pig manure water strain Degradation rate of odor molecules in chicken manure water (%) Degradation rate of odor molecules in pig manure water (%) control group 35.88 5.11 Comamonas testis ketone C1-2/new 100 99.85 Rhodococcus D2-3 100 99.89 Alcaligenes faecalis DO-BC3-1 100 97.53 Acinetobacter Thomsonii DO-Mix-7-1 95.43 100 Contaminating Bacillus brevis G08 99.13 98.75

As shown in Table 1, no matter for chicken manure water or pig manure water, the five strains isolated from mature compost and obtained by screening all have good odor molecule degradation ability, and can degrade more than 95% of odor. molecular.

2. Mixed strain deodorization test

Next, the 5 strains screened out above were individually cultured with 5 ml of tryptic soybean culture medium to reach a stable stage, and the cells were collected by centrifugation, rinsed with reverse osmosis water and centrifuged twice at low speed, and redissolved in 1 ml. of reverse osmosis water. Take 0.5ml of each bacterial solution and transfer it to a shake flask of 50mL of sterilized chicken manure water (ammonia nitrogen value of 1586mL/L) or sterilized pig manure water (ammonia nitrogen value of 1938mL/L) in the same group. After 12 days of shaking culture at 30°C, headspace solid-phase microextraction combined with gas chromatography-mass spectrometry was used to analyze the content of volatile odor molecules in chicken manure water or pig manure water treated with mixed strains for 12 days. The operation steps and subsequent analysis methods of headspace solid-phase microextraction combined with gas chromatography-mass spectrometry are the same as those described in the single-strain deodorization test above, and will not be repeated here.

The degradation rate of odor molecules in chicken manure water or pig manure water by the mixed strains after equal mixing of 5 strains was taken as 100% (that is, the degradation rate 0%), as shown in Table 2 below.

Table 2 - Degradation rate of odor molecules by mixed deodorant strains in chicken manure water or pig manure water group Degradation rate of odor molecules in chicken manure water (%) Degradation rate of odor molecules in pig manure water (%) control group 21.75 6.04 mixed strains 93.34 97.49

As shown in Table 2, whether for chicken manure water or pig manure water, compared with the control group added with reverse osmosis water, the ratios of Comamonas testosterone C1-2/new, Rhodococcus D2-3, Alcaligenes faecalis DO-BC3-1, Acinetobacter thomsonii DO-Mix-7-1 and five strains of Bacillus contaminating Brevibacterium G08 have good odor molecule degradation ability, which can degrade more than 90% odor molecules. It should be noted that, compared with the aforementioned single-strain deodorization test, the chicken manure water and pig manure water used in the mixed-strain deodorization test have higher ammonia nitrogen values, 1586 mL/L and 1938 mL/L, respectively. It can be seen that even if the ammonia nitrogen value of the chicken manure water and the pig manure water in the mixed strain deodorization test increases, the mixed strain used in the present invention can still effectively degrade the odor molecules in the chicken manure water or the pig manure water.

3. Single-strain and mixed-strain small-scale compost deodorization test

In order to be closer to practical application, a small-scale compost deodorization test was used to evaluate the deodorization ability of the above 5 strains, and further compare the compost deodorization ability of a single strain and 5 mixed strains. First, 1kg of chicken manure as an organic material source and 0.45kg of rice husks as a substrate were thoroughly mixed to form a compost with a carbon-nitrogen ratio of about 30 and a moisture content of about 50%, and 50g of the compost was dispensed into 1L in serum bottles. The 5 strains screened out were individually cultured with 50ml of tryptic soybean culture medium to reach the stable stage, and the cells were collected by centrifugation. After washing with reverse osmosis water and low-speed centrifugation twice, they were redissolved in 10ml reverse osmosis. in water. Take an appropriate amount of bacterial liquid to measure OD _600nm and carry out plate medium cultivation by serial dilution method to calculate the bacterial liquid concentration. The measurement results of the OD _600nm and bacterial liquid concentration of each strain are shown in Table 3 below.

Table 3 - Concentration of deodorizing bacteria strain OD _600nm Bacterial concentration (CFU/ml) Comamonas testis ketone C1-2/new 14.44 2.03x10 ¹³ Rhodococcus D2-3 25.62 5.0x10 ¹³ Alcaligenes faecalis DO-BC3-1 11.12 8.8x10 ¹² Acinetobacter Thomsonii DO-Mix-7-1 20.66 6.9x10 ⁹ Contaminating Bacillus brevis G08 6.43 9.3x10 ¹⁰

For the single strain group, 0.5ml of bacterial liquid and 2.0ml of reverse osmosis water were transferred to the serum bottle containing compost, while the mixed strain group each took 0.5ml of a single strain and a total of 2.5ml of bacterial liquid was transferred to the compost-containing serum bottle. 2.5ml of reverse osmosis water was added to the serum bottle containing compost for the control group.

Compost and deodorize by standing at room temperature for 10 days. Finally, the ammonia gas concentration in the serum bottle was measured with an ammonia gas detection tube to evaluate the deodorization ability of single strain and mixed strain on compost. The ammonia concentration measured on day 10 of the small-scale compost deodorization test is shown in Figure 1.

As shown in Figure 1, after 10 days of composting and fermentation, the compost from Comamonas testosterone C1-2/new, Rhodococcus D2-3, Alcaligenes faecalis DO-BC3-1, Thompsonella The mixed strain of Acinetobacter DO-Mix-7-1 and five strains of Bacillus contaminating Brevibacterium G08 can maintain the ammonia content of compost at about 150PPM. Compared with a single strain, the mixed strain formed by 5 strains can still show good deodorization ability in the small-scale compost deodorization test.

4. 100kg scale compost deodorization test

Next, it was further tested whether the mixed strains still have the ability to degrade odor molecules for large-scale composting. 200 kg of chicken manure is thoroughly mixed with 90 kg of coarse bran to form a compost with a carbon-nitrogen ratio of about 30 and a moisture content of about 50%. The mixed composts were divided into two groups, one group was the control group and the other group was the mixed strain experimental group. The 5 strains screened out were individually cultured with 1.5L of tryptic soybean culture medium to reach the stable period, and 0.4L of the bacterial liquid was mixed into 2L mixed strains for subsequent composting and deodorization tests. On the other hand, the control The group used 2L of reverse osmosis water.

Apply 2L of the above-mentioned mixed strains to the compost to form a compost-strain mixture. Next, the compost-strain mixture was turned for the first time. The compost mixture was allowed to stand at room temperature to allow the mixed strains in the microbial deodorant preparation to ferment and decompose odor molecules produced by chicken manure. After the first turn, the compost mix was turned a second time at a frequency of 3 days. At the same time, 2 L of the mixed strain was added.

During the compost deodorization test, the content of odor-contributing odor molecules such as methylamine, dimethylamine and cresol, etc., were analyzed by the detector tube. The sampling time points are 0, 3, 24, 48 and 72 hours after each turn over. Each sampling is covered with a 30L bucket above the compost, and after standing for 10 minutes, the gas in the bucket is collected with an air bag, and The content of odor molecules in the gas collection bag is analyzed by individual detection tubes. The changes in the content of odor molecules in the 100kg-scale compost deodorization test are shown in Figures 2A to 2B, 3A to 3B, and 4A to 4B.

Figure 2A is a graph showing the change of methylamine concentration in the 100kg scale compost deodorization test, after the compost was treated with mixed strains and after the first turning. The concentration of methylamine in the mixed strain experimental group and the control group both reached the highest value within 24 hours after the compost was turned for the first time. Compared with the control group, the methylamine concentration in the experimental group treated with mixed strains decreased by about 33%. . Figure 2B is a graph showing the change of methylamine concentration in the 100kg scale compost deodorization test after the compost was treated with mixed strains and after the second turning. After the second turn over, the concentration of methylamine still reached the highest value at 24 hours, and the concentration of methylamine in the mixed strain experimental group was reduced by about 95% compared with the control group.

Figure 3A is a graph showing the change of dimethylamine concentration in the 100kg scale compost deodorization test after the compost was treated with mixed strains and after the first turning. The concentration of dimethylamine in the mixed strain experimental group and the control group both reached the highest value within 24 hours after the compost was turned for the first time. 30%. Figure 3B is a graph showing the change of dimethylamine concentration in the 100kg scale compost deodorization test after the compost was treated with mixed strains and after the second turning. After the second turn over, the concentration of dimethylamine still reached the highest value at 24 hours, and the concentration of dimethylamine in the mixed strain experimental group was reduced by about 95% compared with the control group.

Figure 4A is a graph showing the change of cresol concentration in the 100kg scale compost deodorization test, after the compost was treated with mixed strains and after the first turning. In Figure 4A, the concentration of cresol in the control group reached the highest value at 72 hours after the first turning of the compost, while the concentration of the mixed strain experimental group remained within 72 hours after the first turning of the compost. Cresol concentrations below about 5PPM. Figure 4B is a graph showing the change of cresol concentration in the 100kg scale compost deodorization test, after the compost was treated with mixed strains and after the second turning. After the second turn over, the cresol concentration of the mixed strain experimental group and the control group reached the highest value at 24 hours, and the cresol concentration of the mixed strain experimental group was reduced by about 28% compared with the control group.

From the results of the 100kg-scale compost deodorization tests shown in Figures 2A to 2B, 3A to 3B, and 4A to 4B, it can be seen that the mixed strains still have the ability to deodorize large-scale composts. The differences in the concentrations of methylamine, dimethylamine and cresol in the mixed strain experimental group and the control group indicated that the mixed strain could be degraded into odor molecules that originated from the odor of biological waste. In addition, according to the results of the 100kg scale compost deodorization test, the concentration of the above-mentioned odor molecules will gradually increase after turning over. As mentioned above, as far as the composting process is concerned, increasing the frequency of composting can accelerate the ripening of the compost, but odors are easily dissipated during the composting process, which will cause environmental pollution. Using the mixed strain of the present invention to treat compost and turning the compost at specific intervals can not only maintain the activity of the mixed strain during the fermentation process, but also can effectively reduce the odor molecules generated by the compost after turning over, and reduce the composting process. pollution to the environment.

Although the present invention has been disclosed above with preferred embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make some changes and modifications without departing from the spirit and scope of the present invention. Therefore, the present invention The scope of protection shall be determined by the scope of the appended patent application.

none.

Figure 1 is a bar graph showing the ammonia concentration of compost treated with a single strain or a mixed strain. Figure 2A is a graph showing the change of methylamine concentration in the 100kg scale compost deodorization test, after the compost was treated with mixed strains and after the first turning. Figure 2B is a graph showing the change of methylamine concentration in the 100kg scale compost deodorization test after the compost was treated with mixed strains and after the second turning. Figure 3A is a graph showing the change of dimethylamine concentration in the 100kg scale compost deodorization test after the compost was treated with mixed strains and after the first turning. Figure 3B is a graph showing the change of dimethylamine concentration in the 100kg scale compost deodorization test after the compost was treated with mixed strains and after the second turning. Figure 4A is a graph showing the change of cresol concentration in the 100kg scale compost deodorization test, after the compost was treated with mixed strains and after the first turning. Figure 4B is a graph showing the change of cresol concentration in the 100kg scale compost deodorization test, after the compost was treated with mixed strains and after the second turning.

Domestic deposit information 1. Comamonas testosteroni C1-2/new ( Comamonas testosteroni C1-2/new) Bioresource Conservation and Research Center, Research Institute of Food Industry Development, Republic of China, deposit number BCRC 911003 on May 26, 2020 2 . Rhodococcus sp. D2-3 ( Rhodococcus sp. D2-3) Bioresource Conservation and Research Center, Food Industry Development Research Institute of the Republic of China, deposit number BCRC 911001 on May 26, 2020 3. Alcaligenes faecalis DO-BC3-1 ( Alcaligenes faecalis DO-BC3-1) Bioresource Conservation and Research Center of the Food Industry Development Research Institute of the Republic of China on May 26, 2020, the deposit number is BCRC 911000 4. Acinetobacter towneri DO-Mix-7-1 ( Acinetobacter towneri DO- Mix-7-1) Bioresource Conservation and Research Center, Research Institute of Food Industry Development of the Republic of China, deposit number BCRC 911002 on May 26, 2020 Deposit and Research Centre 26 May 2020 Deposit number BCRC 911004

Claims (20)

A microbial deodorant preparation, comprising a mixed strain comprising a first group of strains, wherein the first group of strains is selected from Rhodococcus sp. and Acinetobacter towneri At least one of them, and wherein the Rhodococcus is deposited in the Food Industry Development Research Institute of the Republic of China Consortium under the deposit number BCRC 911001, and the Acinetobacter Thomnii is deposited in the Food Industry Development Research Institute of the Republic of China Consortium, under the deposit number BCRC 911001. Number BCRC 911002. The microbial deodorant preparation as claimed in claim 1, wherein the mixed strain further comprises a second group of strains selected from Comamonas testosteroni , Alcaligenes faecalis At least one of Alcaligenes faecalis and Brevibacillus invocatus . The microbial deodorant preparation according to claim 2, wherein the Comamonas testosterone is deposited in the Food Industry Development Research Institute of the Republic of China Foundation, and the deposit number is BCRC 911003. The microbial deodorant preparation according to claim 2, wherein the Alcaligenes faecalis is deposited in the Food Industry Development Research Institute of the Republic of China Foundation, and the deposit number is BCRC 911000. The microbial deodorizing preparation according to claim 2, wherein the contaminating Brevibacillus is deposited in the Food Industry Development Research Institute of the Republic of China Foundation, and the deposit number is BCRC 911004. The microbial deodorizing preparation according to claim 1, wherein the microbial deodorizing preparation is in the form of a suspension or a powder. The microbial deodorizing preparation as claimed in claim 2, wherein: the Comamonas testis ketone is deposited in the Food Industry Development Research Institute of the Republic of China Foundation, and the deposit number is BCRC 911003; the Alcaligenes faecalis is deposited in the Republic of China The Food Industry Development Research Institute, a consortium, has a deposit number of BCRC 911000; The microbial deodorant preparation as claimed in claim 7, wherein the mixed strains of the Comamonas testosterone, the Rhodococcus, the Ralcaligenes faecalis, the Acinetobacter tannerii and the bacteria that contaminate the Brevibacillus The quantity ratio is 1:0.5-1.5:0.5-1.5:0.5-1.5:0.5-1.5. The microbial deodorant preparation as claimed in claim 7, wherein the total bacterial counts of the Comamonas testosterone, the Rhodococcus, the Ralcaligenes faecalis, the Acinetobacter tonneii and the Brevibacillus contamination are between Range between 10 ⁸ cfu/ml to 10 ¹³ cfu/ml. A method for deodorizing compost, comprising the steps of: (a) mixing a biological waste with a substrate to form a compost having a carbon to nitrogen ratio between 30 and 40; (b) applying to the compost as requested The microbial deodorizing preparation of item 1 to form a compost mixture; (c) subjecting the compost mixture to a first turn; and (d) allowing the compost mixture to stand for up to 14 days to allow the mixed strain to ferment the compost and decompose odor molecules produced by the biological waste. The method for deodorizing compost according to claim 10, wherein the substrate comprises wood chips, chaff, straw, bark, fallen leaves, soybean meal, mushroom feet, bagasse, vegetable residue, peanut shells or a combination thereof. The method for deodorizing compost as claimed in claim 10, wherein the odor molecules include ammonia, methylamine, phenol, cresol, indole, derivatives of the foregoing or combinations thereof . The deodorizing method for composting according to claim 10, wherein the microbial deodorizing preparation is in the form of a suspension. The method for deodorizing compost according to claim 13, wherein the applied amount of the microbial deodorizing preparation is 5ml to 30ml per kilogram of the compost. The method for deodorizing compost as claimed in claim 10, further comprising: (c') after step (c), applying the same amount of the microbial deodorizing agent to the compost mixture at an interval frequency, And turn over the compost mixture. The method for deodorizing compost as claimed in claim 15, wherein the interval is between 0.5 days and 5.5 days. The method for deodorizing compost as claimed in claim 16, wherein the interval is between 2.5 days and 3.5 days. Use of a mixed strain for compost deodorization, wherein the mixed strain includes a first group of strains, the first group of strains being selected from Rhodococcus and Thompsonii At least one of Acinetobacter, and wherein the Rhodococcus is deposited in the Food Industry Development Research Institute of the Republic of China Consortium, the deposit number BCRC 911001, and the Acinetobacter tonneii is deposited in the Food Industry Development Research Institute of the Republic of China Consortium , deposit number BCRC 911002. Use of a mixed strain for composting deodorization, wherein the mixed strain is composed of Comamonas testosterone, Rhodococcus, Alcaligenes faecalis, Acinetobacter tanninii, and Brevibacterium contamination, and wherein the red The Coccus genus is deposited in the Food Industry Development Research Institute of the Republic of China Consortium, and the deposit number is BCRC 911001. The use as described in claim 19, wherein: the Comamonas testosterone is deposited in the Food Industry Development Research Institute of the Republic of China Consortium, and the deposit number is BCRC 911003; the Alcaligenes faecalis is deposited in the Food Industry of the Republic of China Consortium Industrial Development Research Institute, deposit number BCRC 911000; the Acinetobacter tangmanii line is deposited in the Food Industry Development Research Institute of the Republic of China Consortium, deposit number BCRC 911002; and the Brevibacillus contamination is deposited in the Food Industry Development Research Institute of the Republic of China Consortium So, deposit number BCRC 911004.

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