CN107022494B - Dictyophora pinicola BN-h1 and biological bacterium degradation agent as well as preparation method and application thereof - Google Patents
Dictyophora pinicola BN-h1 and biological bacterium degradation agent as well as preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to a dimoxysporum pinellia BN-h1 strain and a biological bacterium degradation agent containing dimoxysporum pinellia BN-h1, and a preparation method and application thereof, wherein the dimoxysporum pinellia BN-h1 is preserved in the China general microbiological culture Collection center of the Committee for culture Collection of microorganisms with the preservation number of CGMCC No.13727 and the preservation date of 2017, 3 months and 7 days. The invention can degrade ammonia nitrogen with the maximum concentration of 1200mg/L by using the bicellular pine BN-h1, and meanwhile, the degradation efficiency of the ammonia nitrogen can reach more than 90 percent.
Description
Technical Field
The invention relates to the field of fungi, and particularly relates to a strobilus Pini and a biological bacterium degradation agent, and a preparation method and application thereof.
Background
The municipal refuse sanitary landfill is the main mode of centralized disposal of municipal refuse in China at present, and a large amount of percolate can be generated in a refuse landfill during the service life until the refuse is scrapped. In recent years, along with the rapid development of economy in China, the landfill leachate has larger output and higher ammonia nitrogen content, and becomes one of the environmental problems concerned by students. The content of ammonia nitrogen is an important index of the quality of sewage, is the main form of nitrogen in a water-phase environment, and is one of main factors causing eutrophication of a water body, and the high-concentration ammonia nitrogen can reduce the dissolved oxygen in water, lead fishes to die in large quantities, even lead lakes to dry and die, bring great loss to agricultural production and aquaculture, so that how to effectively degrade the content of ammonia nitrogen in the water body and not cause secondary pollution become a hotspot of research of people. The degradation of ammonia nitrogen by using microorganisms is a mature method among a plurality of methods, and the method utilizes the principle of 'nitrification-denitrification' and the interaction of nitrosobacteria and nitrobacteria.
The traditional theory holds that the nitrification reaction and the denitrification reaction are two different biological reaction processes, and the nitrification reaction refers to the reaction of NH under the action of microorganisms4 + Oxidation of-N to NO2 - -N (nitrosation) is further oxidized to NO3 - -N (nitration) process, the denitrification reaction being NO3 - Reduction of-N to N by microbial action2The nitrification is carried out under aerobic conditions and the denitrification is carried out under anaerobic conditions.
The sewage treatment technology generally adopts a physical treatment method, a chemical treatment method, a biochemical treatment method or a biological treatment method, but more than 90 percent of sewage treatment plants in various countries in the world since the 19 th century adopt the biological treatment technology, because the microbial degradation of the overhigh ammonia nitrogen content in the water body has the advantages which are incomparable with the traditional method, the treatment efficiency is higher, the secondary pollution is not generated, the required instruments are simple and easy to operate, and the technology has great development potential and application prospect.
Disclosure of Invention
The invention aims to culture the bicella matsutake capable of degrading high ammonia nitrogen, and the bicella matsutake can degrade the ammonia nitrogen with the concentration of 1200mg/L and simultaneously ensure that the ammonia nitrogen degradation efficiency reaches more than 90 percent. The purpose of the invention is realized by the following technical scheme.
According to one aspect of the invention, the inventor selects and breeds a fungus strain through screening. Through comparison, the developing tree of the fungus has the highest sequence homology with the registered Erodia pinicola (Diplodia pinea) to reach 100 percent, so that the fungus can be determined to be a Erodia pinicola strain named as the Erodia pinicola (Diplodia pinea) BN-h1, which is currently preserved in the China general microbiological culture Collection center (CGMCC for short). The address of the preservation unit is No. 3 of Xilu No.1 of Beijing, China, rising area, the microbial research institute of Chinese academy of sciences, the preservation number is CGMCC No.13727, and the preservation date is 3 months and 7 days in 2017. The bicellular pine chrophilum is a microbial inoculum capable of degrading ammonia nitrogen in sewage and leachate and ammonia gas in odor, can effectively degrade the ammonia nitrogen with the concentration of 0-1200 mg/L, and can simultaneously carry out nitration and denitrification reactions under aerobic conditions to degrade the ammonia nitrogen.
According to a second aspect of the present invention, there is provided a biological bacteria degrading agent comprising dicentra pinea BN-h 1.
According to a third aspect of the present invention, there is provided a method for preparing a biological bacteria degradation agent, comprising the steps of:
(1) inoculating a single colony of the bicellular pine BN-h1 into a seed culture medium for culture to prepare a biological bacteria degradation agent mother solution.
(2) Adding the mother liquor into a fermentation culture medium for fermentation culture to prepare the biological bacteria degradation agent.
Wherein, the seed culture medium comprises the following components: 19.02g/L brown sugar, 8.17g/L sodium citrate, K2HPO414g/L，KH2PO46g/L，(NH4)2SO44.72g/L，MgSO4·7H20.2g/L of O, 2mL/L of trace elements and the balance of water.
The fermentation medium comprises the following components: 19.02-57.06 g/L brown sugar, 8.17-24.51 g/L sodium citrate, K2HPO414g/L，KH2PO46g/L，(NH4)2SO44.72～14.16g/L，MgSO4·7H20.2g/L of O, 2mL/L of trace elements and the balance of water.
Wherein, step (1) includes: the culture temperature is 25-35 ℃, the initial pH value of the culture medium is 6.5-8.5, the rotation speed is 150-200 rpm, and the fermentation time is 72 hours.
The step (2) comprises the following steps: the fermentation temperature is 25-35 ℃, the initial pH value of the fermentation medium is 6.5-8.5, the rotation speed is 150-200 r/min, and the fermentation culture lasts for 36-60 hours.
Wherein the C/N ratio of the seed culture medium to the fermentation culture medium is 9-11: 1, and the high-efficiency degradation can be achieved under the ratio.
The ammonium sulfate is the only nitrogen source of the culture medium, the brown sugar and the sodium citrate are both carbon sources of the culture medium, and when the adding ratio of the brown sugar to the sodium citrate is 3-5: 1, the ammonia nitrogen degradation effect is optimal. The combination of brown sugar and sodium citrate also has an adjusting effect on the pH value of the culture medium on the basis of providing a carbon source, so that the pH values of the enrichment culture medium, the separation culture medium, the seed culture medium and the fermentation culture medium are all 7.0-8.0.
Wherein, the preparation method also comprises a separation and purification step of the bicyces pinocyani BN-h1 before the step (1), and the steps are as follows:
the composition of the enrichment medium is: 19.02-29.68 g/L of brown sugar, K2HPO414g/L，KH2PO46g/L，(NH4)2SO44.72～6g/L，MgSO4·7H20.2g/L of O, 2mL/L of trace elements and the balance of water.
The composition of the isolation medium was: 19.02-29.68 g/L of brown sugar, K2HPO414g/L，KH2PO46g/L，(NH4)2SO44.72～6g/L，MgSO4·7H20.2g/L of O, 2mL/L of trace elements, 20g/L of agar and the balance of water.
Wherein, the composition of the trace elements is as follows: EDTA 2Na 5-57.1 g/L, ZnSO4·7H2O 3.9g/L，CaCl2·2H2O 7g，MnCl2·4H2O 5.1g/L，FeSO4·7H2O 5g/L，(NH4)6Mo7O24·4H2O1.1g/L，CuSO4·5H2O1.6g/L，CoCl2·6H2O1.6 g/L, and the balance of water.
According to a fourth aspect of the present invention there is provided the use of a strobilus Pini BN-h1 or a biological bacteria degrading agent in the remediation of sewage, leachate and odor pollution.
The invention has the beneficial effects that:
in the process of degrading ammonia nitrogen, the bicellular turquoise BN-h1 has no accumulation of nitrite nitrogen and nitrate nitrogen, can synchronously carry out nitrification and denitrification, and has high degradation rate of the ammonia nitrogen up to more than 90 percent.
Drawings
Various other advantages and benefits will become apparent to those of ordinary skill in the art upon reading the following detailed description of the preferred embodiments. The drawings are only for purposes of illustrating the preferred embodiments and are not to be construed as limiting the invention. Also, like reference numerals are used to refer to like parts throughout the drawings. In the drawings:
FIG. 1 shows a picture of the culture morphological characteristics of a strain of Dictyophora pinea BN-h1 according to an embodiment of the invention;
FIG. 2 shows a picture of a developmental tree of a strain of Dictyocaulus pinicola BN-h1 according to an embodiment of the invention.
Detailed Description
Exemplary embodiments of the present disclosure will be described in more detail below with reference to the accompanying drawings. While exemplary embodiments of the present disclosure are shown in the drawings, it should be understood that the present disclosure may be embodied in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the disclosure to those skilled in the art.
The screening method obtains the Dictyophora loose (BN-h 1) for degrading high-concentration ammonia nitrogen in sewage and percolate and high-concentration ammonia gas in odor, the ammonia nitrogen degradation capability of the Dictyophora loose (BN-h 1) reaches more than 90%, the capability of the strain and a biological bacteria degradation agent containing the strain on treating polluted sewage and percolate with high ammonia nitrogen content and odor is better, the odor and odor pollution of the sewage and percolate can be effectively solved, and the contribution is made to environmental treatment.
The bicellular pinocytosporum BN-h1 strain disclosed by the invention has the following main characteristics:
culture morphological characteristics of bicellular pine I-B strain BN-h1
As shown in FIG. 1, the Diacocephalum pini BN-h1 has the following morphological characteristics: milky white is yellowish, the growth speed on a flat plate is high, the protrusions grow, the edges are neat, thalli are wet and smooth, the front and back colors are the same, the thalli are easy to pick out, the growth in a liquid culture medium is high, and the culture medium is turbid.
Developmental tree of Blueslea arborea BN-h1
The MEGA5.0 is used as a development tree, and the sequence homology of the gene sequence of the BN-h1 and the registered Erysia pinicola (Diplodianea) can reach 100 percent, so that the Erysia pinicola is determined and is shown in a specific figure 2.
ITS sequence of Blastomyces tersii BN-h1
Selecting a single bacterial colony of purified BN-h1, inoculating the single bacterial colony in a seed culture medium, carrying out shaking culture for 24 hours at the temperature of 30 ℃ under the condition of 180 revolutions per minute of a shaking table, then using a bacterial liquid as a template to carry out ITS sequence of a PCR amplification strain, carrying out sequencing, and carrying out sequence comparison on a http:// archive-dtd. The ITS sequence of BN-h1 is specifically shown in SEQ ID NO 1.
The invention also relates to a preparation method of the biological bacteria degradation agent containing the strobilus Pini (Diplodia pinea) BN-h1, which specifically comprises the separation and purification process and the culture fermentation process of the strobilus Pini (Diplodia pinea) BN-h1, wherein the separation and purification process scheme is as follows:
taking 10ml of landfill leachate, inoculating the landfill leachate into a 250ml triangular flask filled with 100ml of sterilized enrichment medium, shaking up fully, carrying out shake culture at the temperature of 28 ℃ and the rotating speed of 160 r/min, enriching for 7 days, and supplementing (NH) every 1 day4)2SO4To eliminate the unavailable NH4 +-N. Spreading 1ml of the bacteria liquid in the enrichment medium on a separation medium plate, and inversely culturing in an incubator at 28 DEG CCulturing, selecting single colony growing on the plate, and repeatedly separating and purifying to obtain a single strain named as BN-h 1.
The scheme for preparing the biological bacteria degradation agent by culturing and fermenting single bacterial colonies of the Diplodia pine BN-h1 is as follows:
selecting a single bacterial colony of purified BN-h1, inoculating the single bacterial colony into a seed culture medium, controlling the temperature to be 25-35 ℃, adjusting the pH to be 6.5-8.5, rotating the speed to be 150-200 rpm, fermenting for 72 hours to prepare a biological bacteria degradation agent mother liquor, adding the mother liquor into the fermentation culture medium according to the proportion of 1%, controlling the temperature to be 25-30 ℃, the pH to be 6.5-8.5, rotating the speed to be 150-200 rpm, and fermenting for 36-60 hours to prepare the biological bacteria degradation agent.
The present invention, namely, Diplodicoccus pinicola BN-h1 (or a biodegradation agent containing the same), and the degradation function thereof will be further described below by way of specific examples.
Example 1
Isolation and purification scheme 1 of Dictyota pine (Diplodia pinea) BN-h 1:
(1) taking 10ml of refuse leachate from Nangong refuse composting farm in Beijing City, inoculating into 250ml triangular flask containing 100ml of sterilized enrichment medium, shaking thoroughly, performing shake culture at 28 deg.C and 160 rpm, enriching for 7 days, and supplementing every 1 day (NH)4)2SO4To eliminate the unavailable NH4 +-N.
(2) And (3) coating 1ml of the bacterial liquid in the enrichment medium on a separation medium plate, carrying out inverted culture in an incubator at 28 ℃, selecting a single bacterial colony growing on the plate, and repeatedly separating and purifying to obtain a single bacterial colony of the Diplodia pine BN-h 1.
Wherein, the enrichment medium comprises the following components: brown sugar 19.02g/L, K2HPO414g/L，KH2PO46g/L，(NH4)2SO44.72g/L，MgSO4·7H20.2g/L of O, 2mL/L of trace elements and the balance of water.
The composition of the isolation medium was: brown sugar 19.02g/L, K2HPO414g/L，KH2PO46g/L，(NH4)2SO44.72g/L，MgSO4·7H20.2g/L of O, 2mL/L of trace elements, 20g/L of agar and the balance of water.
And (3) degradation function detection:
the ammonia nitrogen index determination is carried out on a sample taken back by a big tower of a Nanogong biological deodorization tower in Beijing: the ammonia nitrogen content of the tower odor washing liquid is 1184mg/L, the pH value is 6.47, and the ammonia gas content at the gas outlet is 8.34mg/m3(ii) a Adding the obtained BN-h1 strain and odor washing liquid in the tower in a proportion of 1% into a south America big tower, culturing for 10 days in an aeration environment at 32 ℃, degrading ammonia nitrogen in the odor washing liquid of the big tower to 133.23mg/L, wherein the pH value is 7.16, and the ammonia gas content at an air outlet is 1.76mg/m3。
Example 2
Culture protocol for biodegradation agent 1:
(1) taking 10ml of garbage percolate of the Beijing south palace garbage composting farm, inoculating the garbage percolate into a 250ml triangular flask filled with 100ml of sterilized enrichment medium, fully shaking up, carrying out shake culture at the temperature of 28 ℃ and the rotating speed of 160 r/min, enriching for 7 days, and supplementing every 1 day (NH)4)2SO4To eliminate the unavailable NH4 +-N.
(2) And (3) coating 1ml of the bacterial liquid in the enrichment medium on a separation medium plate, carrying out inverted culture in a 28 ℃ incubator, and selecting a single bacterial colony growing on the plate to carry out repeated separation and purification to obtain a single bacterial colony of BN-h 1.
(3) And selecting a single bacterial colony of the purified BN-h1, inoculating the single bacterial colony into a seed culture medium, controlling the temperature to be 25 ℃, adjusting the pH to be 6.5, rotating the speed to be 150rpm, and fermenting for 72 hours to prepare the mother liquor of the biological bacteria degradation agent.
(4) Adding the mother liquor into a fermentation culture medium according to the proportion of 1%, controlling the temperature to be 35 ℃, the pH value to be 6.5, the rotating speed to be 200rpm, and the fermentation time to be 60 hours, and preparing the biological bacteria degradation agent.
The composition of the enrichment medium is: brown sugar 29.68g/L, K2HPO414g/L，KH2PO46g/L，(NH4)2SO46g/L，MgSO4·7H20.2g/L of O, 2mL/L of trace elements and the balance of water.
The composition of the isolation medium was: brown sugar 29.68g/L, K2HPO414g/L，KH2PO46g/L，(NH4)2SO46g/L，MgSO4·7H20.2g/L of O, 2mL/L of trace elements, 20g/L of agar and the balance of water.
The seed culture medium comprises the following components: 19.02g/L brown sugar, 8.17g/L sodium citrate, K2HPO414g/L，KH2PO46g/L，(NH4)2SO44.72g/L，MgSO4·7H20.2g/L of O, 2mL/L of trace elements and the balance of water.
The fermentation medium comprises the following components: 19.02g/L brown sugar, 8.17g/L sodium citrate, K2HPO414g/L，KH2PO46g/L，(NH4)2SO44.72g/L，MgSO4·7H20.2g/L of O, 2mL/L of trace elements and the balance of water.
And (3) degradation function detection:
the ammonia nitrogen index determination is carried out on a sample taken back by a small tower of a Nanogong biological deodorization tower in Beijing: the ammonia nitrogen content of the tower odor washing liquid is 1471mg/L, the pH value is 6.38, and the ammonia gas content at the gas outlet is 9.13mg/m3(ii) a Adding biological bacteria degradation agent 1 into small south China cell tower at a ratio of BN-h1 and odor washing liquid in the tower of 1%, culturing for 10 days in aeration environment at 32 ℃, wherein the ammonia nitrogen degradation efficiency of the odor washing liquid in the small south China cell tower is over 90%, the pH value is 7.23, and the ammonia content at the gas outlet is 2.25mg/m3。
Example 3
Culture protocol of biodegradation agent 2:
(1) taking 10ml of garbage percolate of Ashuec garbage composting farm in Beijing, inoculating into a 250ml triangular flask containing 100ml of sterilized enrichment medium, shaking up sufficiently, performing shake culture at 28 deg.C and 160 rpm, enriching for 7 days, and supplementing every 1 day (NH)4)2SO4To eliminate the unavailable NH4 +-N.
(2) And (3) coating 1ml of the bacterial liquid in the enrichment medium on a separation medium plate, carrying out inverted culture in a 28 ℃ incubator, and selecting a single bacterial colony growing on the plate to carry out repeated separation and purification to obtain a single bacterial colony of BN-h 1.
(3) And selecting a single bacterial colony of the purified BN-h1, inoculating the single bacterial colony into a seed culture medium, controlling the temperature to be 35 ℃, adjusting the pH to be 6.5, rotating the speed to be 150rpm, and fermenting for 72 hours to prepare the mother liquor of the biological bacteria degradation agent.
(4) Adding the mother liquor into a fermentation culture medium according to the proportion of 1%, controlling the temperature to be 25 ℃, the pH value to be 8.5, the rotating speed to be 150rpm, and the fermentation time to be 36 hours, and preparing the biological bacteria degradation agent.
The composition of the enrichment medium is: brown sugar 29.68g/L, K2HPO414g/L，KH2PO46g/L，(NH4)2SO46g/L，MgSO4·7H20.2g/L of O, 2mL/L of trace elements and the balance of water.
The composition of the isolation medium was: brown sugar 19.02g/L, K2HPO414g/L，KH2PO46g/L，(NH4)2SO46g/L，MgSO4·7H20.2g/L of O, 2mL/L of trace elements, 20g/L of agar and the balance of water.
The seed culture medium comprises the following components: 19.02g/L brown sugar, 8.17g/L sodium citrate, K2HPO414g/L，KH2PO46g/L，(NH4)2SO44.72g/L，MgSO4·7H20.2g/L of O, 2mL/L of trace elements and the balance of water.
The fermentation medium comprises the following components: 57.06g/L brown sugar, 24.51g/L sodium citrate, K2HPO414g/L，KH2PO46g/L，(NH4)2SO414.16g/L，MgSO4·7H20.2g/L of O, 2mL/L of trace elements and the balance of water.
And (3) degradation function detection:
index measurement is carried out on water samples taken back by odor washing liquid of a biological deodorization tower in an Ashbya waste composting farm in Beijing and ammonia gas at an air inlet and an air outlet collected by the biological deodorization towerDetermining: the ammonia nitrogen content of the odor washing liquid is 1500mg/L, the pH value is 7.45, and the ammonia gas content at the air inlet of the biological deodorization tower is 23.27mg/m3The ammonia gas content at the gas outlet is 13.66mg/m3(ii) a Adding biological bacteria degradation agent 2 into odor cleaning solution of Ashbya deodorizing tower at a ratio of BN-h1 to sewage in the tower of 1%, culturing at 30 deg.C in aeration environment for 10 days until ammonia nitrogen is degraded to 129mg/L, the degradation efficiency reaches above 90%, pH value is 7.26, and ammonia content at air inlet is 21.28mg/m3However, the ammonia gas content at the gas outlet was 1.21mg/m3。
By combining the embodiment and the characteristics of the bicellular pine BN-h1, the bicellular pine BN-h1 disclosed by the invention has the advantages that high-concentration ammonia nitrogen can be degraded, the sewage stink and stink pollution caused by ammonia nitrogen can be effectively relieved, the treatment time in production is short, and the application effect is obvious.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are included in the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the appended claims.
SEQUENCE LISTING
<110> Beijing epoxy environmental protection science and technology development Co Ltd/Beijing college of agriculture
<120> a strain of bicellular pine BN-h1, biological bacterium degradation agent, and preparation method and application thereof
<160>1
<170>PatentIn version 3.5
<210>1
<211>567
<212>DNA
<213> Trichosporon pine (Diplodia pinea)
<400>1
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atgcgagaac caagagatcc gttgttgaaa gttttgaaga ttaattcaaa atttgactaa 420
ctgtaaaaat aattaaattg tgttttgtta aacctctggc ccaacctatc tctaggccaa 480
accaaagcaa gagttctgta tcaaaaagac actgtgtgta aggtttttcg ccgcgcagtt 540
aagcgctggc aaaagaatac tgtaatg 567
Claims (10)
1. A kind of strobilus PiniDiplodia pinea) BN-h1, which is preserved in the China general microbiological culture Collection center with the preservation number of CGMCC No.13727 and the preservation date of 2017, 3 months and 7 days.
2. A biological bacteria degrading agent comprising the bicellular pine BN-h1 of claim 1.
3. A method of preparing the biological bacteria degrading agent of claim 2, comprising the steps of:
(1) inoculating a single colony of the bicellular pine BN-h1 into a seed culture medium for culture to prepare a biological bacteria degradation agent mother solution;
(2) adding the mother liquor into a fermentation culture medium for fermentation culture to prepare the biological bacteria degradation agent.
4. The method for producing a biological degradation agent according to claim 3,
the seed culture medium comprises the following components: 19.02g/L brown sugar, 8.17g/L sodium citrate, K2HPO414g/L，KH2PO46g/L，(NH4)2SO44.72g/L，MgSO4·7H20.2g/L of O, 2mL/L of trace elements and the balance of water;
the fermentation medium comprises the following components: 19.02-57.06 g/L brown sugar, 8.17-24.51 g/L sodium citrate, K2HPO414g/L，KH2PO46g/L，(NH4)2SO44.72～14.16g/L，MgSO4·7H20.2g/L of O, 2mL/L of trace elements and the balance of water.
5. The method of claim 3, wherein step (1) comprises:
the culture temperature is 25-35 ℃, the initial pH value of the culture medium is 6.5-8.5, the rotating speed is 150-200 rpm, and the fermentation time is 72 hours;
the step (2) comprises the following steps:
the fermentation temperature is 25-35 ℃, the initial pH value of the fermentation medium is 6.5-8.5, the rotation speed is 150-200 r/min, and the fermentation culture lasts for 36-60 hours.
6. The method for producing a biological degradation agent according to claim 4,
the C/N ratio of the seed culture medium to the fermentation culture medium is 9-11: 1.
7. The method for producing a biological degradation agent according to claim 4,
the ratio of brown sugar to sodium citrate is 3-5: 1, and the pH values of the seed culture medium and the fermentation culture medium are 7.0-8.0.
8. The method for preparing a biological degradation agent according to claim 3, wherein the method further comprises a separation and purification step of the bicellular pine BN-h1 before the step (1), wherein the steps of:
the composition of the enrichment medium is: 19.02-29.68 g/L of brown sugar, K2HPO414g/L，KH2PO46g/L，(NH4)2SO44.72～6g/L，MgSO4·7H20.2g/L of O, 2mL/L of trace elements and the balance of water;
the composition of the isolation medium was: 19.02-29.68 g/L of brown sugar, K2HPO414g/L，KH2PO46g/L，(NH4)2SO44.72～6g/L，MgSO4·7H20.2g/L of O, 2mL/L of trace elements, 20g/L of agar and the balance of water.
9. The method for preparing a biological bacteria degrading agent according to any one of claims 4, 6, 7 and 8, wherein the composition of the trace elements is as follows:
EDTA·2Na 5～57.1g/L，ZnSO4·7H2O 3.9g/L，CaCl2·2H2O 7g，MnCl2·4H2O 5.1g/L，FeSO4·7H2O 5g/L，(NH4)6Mo7O24·4H2O 1.1g/L，CuSO4·5H2O 1.6g/L，CoCl2·6H2o1.6 g/L, and the balance of water.
10. Use of the bicellular pine BN-h1 of claim 1 or the biological bacteria degrading agent of claim 2 in treating ammonia nitrogen-containing sewage, leachate and odor pollution.
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