CN111378594B - Compound microbial agent and application thereof - Google Patents

Compound microbial agent and application thereof Download PDF

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CN111378594B
CN111378594B CN201811640134.4A CN201811640134A CN111378594B CN 111378594 B CN111378594 B CN 111378594B CN 201811640134 A CN201811640134 A CN 201811640134A CN 111378594 B CN111378594 B CN 111378594B
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vibrio
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徐军
孙文妮
王开春
田凤蓉
李坤
王强
洪磊
张璐璐
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Bluestar Lehigh Engineering Institute
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Abstract

The invention discloses a compound microbial agent and application thereof. A composite microbial agent contains Vibrio halophilic sp.YL5-2 with preservation number CGMCC NO.16315 and Vibrio halophilic dentrificins DSM15503 with preservation number DSM 15503; or contains the vibrio halovirido sp.YL5-2 bacterial liquid with the preservation number of CGMCC NO.16315 and the bacterial liquid obtained by enrichment culture of seawater under the condition that TDS is more than 10 percent. The microbial agent can be applied to the degradation and transformation of pollutants with the salt concentration of 1-30 percent, and is particularly suitable for industrial scale high-salt wastewater treatment, soil remediation, underground water treatment, marine environment remediation, ecological environment treatment and the like.

Description

Compound microbial agent and application thereof
Technical Field
The invention relates to the field of microorganisms, and relates to a compound microbial agent and application thereof.
Background
Halophilic microorganisms are extreme microorganisms capable of growing and multiplying normally in a high-salt environment, and particularly show great development potential in the aspect of high-salt environment treatment. Common halophiles include Halococus, Halococcus, Halomonas, Halovibrio, Halinicoccus, Marinoccus, Bacillus, Alteromonas, Acinetobacter, Marinomonas, Flavobacterium, Halorusrubrium, Haloferax, Haloarcula, Halobaculum, Natialba, Nationomonas, Natronomas, Natronococcus, etc.
The genus Halovibrio is an important halophilic microorganism, but at present, the genus only has 2 species. Fendrich (Fendrich C. Halovibrio variabilis gen. nov. sp. nov., Pseudomonas halophilia sp. nov. and a new halophilic coccic bacterium from Great Salt Lake, Utah, USA. Syst Appl Microbiol 1988; 11: 36-43.) was isolated from the Great Salt Lake in 1988 to obtain the first Vibrio salinivii haloviridis. Until 2006, Sorokin DY et al (Tourova TP, Galinski EA, Belloch C, Tindall BJ. extreme topographic differentiating bacteria from hypersaline in lanes, Halovibrio densitificans sp.nov. and Halospina densitificans gen.nov., sp.nov., and ovid DYScience that the gene name Halovibrio Ferdrich 1989with the type specs Halovibrio variabil. int J Syst Evol Microbiol 2006; 56: 379-. At present, the Halovibrio genus only comprises two species of Halovibrio variabilis and Halovibrio denitiricican, and the corresponding model bacteria are Halovibrio variabilis DSM3050TAnd Halovibro dentificas DSM15503T。
The separated Halovibrio salina belongs to extreme halophilic bacteria in a high-salt environment, can quickly grow and metabolize under the environment condition that the mass fraction of NaCl is more than 10 percent and even more than 20 percent, and has great application potential in the aspect of environmental pollution control. Screening Halovibrio new strains from a high-salt environment and preparing an environment-friendly microbial agent capable of being applied industrially are one of the technical problems to be solved by environment workers.
Disclosure of Invention
One of the purposes of the invention is to provide a compound microbial agent containing new strains of vibrio salinophilus (Halovibrio).
The invention also aims to provide the composite microbial agent for degrading and converting pollutants under the high-salt condition.
The technical problem to be solved by the invention is realized by the following technical scheme:
a composite microbial agent contains Halovibrio sp.YL5-2 of Vibrio halophilus with preservation number of CGMCC NO. 16315.
The compound microbial agent preferably contains Vibrio halophilic sp.YL5-2 with the preservation number of CGMCC NO.16315 and Vibrio halophilic halioticus DSM15503 with the preservation number of DSM 15503.
As another preferred embodiment of the invention, the compound microbial agent preferably contains a vibrio halovirido sp.YL5-2 bacterial liquid of Vibrio salina with the preservation number of CGMCC NO.16315 and a bacterial liquid obtained by enrichment culture of seawater under the condition that TDS is more than 10%.
The invention discloses a halophilic vibrio halovirido sp.YL5-2 which is preserved in China general microbiological culture collection center with the preservation number of CGMCC NO.16315 and the preservation date of 2018, 8 and 20 days.
The invention discloses a halophilic vibrio halogrio sp.YL5-2 which belongs to a new strain of the halophilic vibrio (halogrio) and proves that the material is as follows:
the accession number of GenBank/EMBL/DDBJ of the 16S rRNA sequence of the Halovibrio salina sp.YL5-2 is MF782425, and the sequence is shown as SEQ ID NO. 1; GenBank/EMBL/DDBJ of the whole genome sequence has the accession number of NSKD 00000000.1.
The 16S rRNA of Halovibrio salina sp.YL5-2 and Halovibrio variabilis DSM3050TThe 16S rRNA similarity of (a) was 97.7%, which is similar to Halovibro dentificas DSM15503TThe similarity of the two groups was 97.3%, which is similar to Halospina dentificans HGD 1-3TThe similarity of (a) was 95.5%, and the level of similarity of the 16S rRNA gene sequence with other types of strains of the Halospina genus was less than 97.0%. The DNA-DNA hybridization values of YL5-2 with Halovibro variabilis DSM3050, Halovibro dentificas DSM15503 were 43.5% and 38.2%, respectively, well below the 70% threshold (the threshold for which the species division is generally accepted).
The Halovibrio salina sp.YL5-2 is gram-negative, aerobic, straight rod-shaped or Vibrio mimicus with the characters of 0.5-0.8 mu m multiplied by 1.0-3.5 mu m, and bacterial colonies on a solid culture medium are smooth and light yellow through the movement of unipolar flagella.
The Vibrio salinophilus Halovibrio sp.YL5-2 can grow in the range of salt concentration of 3% -30%, pH6.5-11.0, and temperature of 15-45 ℃; the optimum growth salt environment is 10-25%, the optimum growth pH is 7.5-8.0, and the optimum growth temperature is 35 ℃.
The main respiratory quinone of Vibrio halophilic Halovibrio sp.YL5-2 is Q-9, the main fatty acids are C18:1 omega 9C, C16:0, C19:0cyclo omega 8C and Summed Feature 8, and the main polar lipids are Diphosphatidylglycerol (DPG), Phosphonomethylethanolamine (PE), Phosphonoglyycerol (PG), Phosphonolvine (PC), which are all the same as or close to the related species Halovibro of YL 5-2.
The Halovibrio salinophilus sp.YL5-2 can utilize bromine-succinic acid, propionic acid and acetic acid as the only carbonSource utilization; but does not utilize D-maltose, D-fructose, D-galactose, D-cellobiose, stachyose, D-melibiose, N-acetyl-D-galactosamine, D-fucose, L-rhamnose, D-mannitol, D-galacturonic acid, D-aspartic acid, D-serine, D-glucuronic acid, L-lactic acid, quinic acid, mucic acid, D-malic acid, gamma-amino-butyric acid, formic acid, acetoacetic acid as the sole carbon source. Halovibrio salina sp. YL5-2 and Halovibrio variabilis DSM3050T、Halovibrio denitrificans DSM 15503TAnd other relatives have obvious difference in physiological and biochemical characteristics.
Based on the differences of phylogenetic analysis of 16SrRNA, genome sequencing, DNA hybridization tests, fatty acid composition, respiratory quinone categories, physiological biochemistry and phenotypic characteristics and the like, the strain YL5-2 can be determined to be a new species of Halovibrio, and is named Halovibrio salipaludis sp. The strain YL5-2 is deposited in the China general microbiological culture Collection center and is Vibrio halophilus Halovibrio sp.
The vibrio salinophilus Halovibrio sp.YL5-2 can degrade pollutants within the range of salt concentration of 3% -32%, and can be used for treating high-salt wastewater with salt concentration of more than 3%, particularly high-salt wastewater with salt concentration of 10% -25%.
The microbial agent can be used for degrading and converting pollutants under the condition of high salinity, and comprises high salinity wastewater treatment, polluted seawater treatment, saline-alkali soil remediation and the like.
The wastewater is preferably epoxypropane wastewater, preserved szechuan pickle wastewater, epichlorohydrin wastewater, polluted underground water in offshore areas and acetic acid polluted wastewater.
The soil is preferably saline-alkali soil, seaport polluted wetland and high-salinity pesticide wastewater polluted soil.
The microbial agent disclosed by the invention is preferably applied to the degradation and transformation of pollutants with the salt concentration of 1-30%, and particularly applied to industrial-scale high-salt wastewater treatment, soil remediation, underground water treatment, marine environment remediation, ecological environment treatment and the like.
The compound microbial agent is applied to the degradation and conversion of pollutants within the range of 1-30% of salt concentration.
The compound microbial agent is preferably applied to the degradation and conversion of high-salinity wastewater pollutants within the range of 10-25% of salt concentration.
The application of the halophilic vibrio Halovibro sp.YL5-2 with the preservation number of CGMCC NO.16315 in wastewater treatment, polluted seawater treatment and soil remediation.
The wastewater is preferably epoxypropane wastewater, preserved szechuan pickle wastewater, epichlorohydrin wastewater, polluted underground water in offshore areas and acetic acid polluted wastewater.
The soil is preferably saline-alkali soil, seaport polluted wetland and high-salinity pesticide wastewater polluted soil.
The application of Halovibrio sp.YL5-2 of Vibrio halophilus with the preservation number of CGMCC NO.16315 in the degradation and transformation of pollutants with the salt concentration of 1-30 percent.
Has the advantages that:
the invention provides a compound microbial inoculum containing YL5-2, which can be applied to the degradation and transformation of pollutants with the salt concentration of 1-30%, in particular to the treatment of high-salt wastewater, soil remediation, groundwater treatment, marine environment remediation, ecological environment treatment and the like on an industrial scale.
Description of the drawings:
FIG. 1 is a photograph of a Transmission Electron Microscope (TEM) of cells of Vibrio halophilus Halovibro sp.YL5-2 cultured on YL agar plates at 37 ℃ for 24 hours (scale bar: 2 μm).
FIG. 2 shows Halovibrio salina sp.YL5-2(a) and Halovibrio dentificans DSM15503T(b)，Halovibrio variabilis DSM 3050T(c) Polar lipid profile of (a).
All polar lipids of the above 3 strains were detected on TLC plates stained with 95% ethanol containing 10% phosphomolybdic acid for total lipids (as shown in the above figure), 1-butanol containing 0.2% ninhydrin for amino lipids (not shown), molybdenum blue reagent (Sigma) for phospholipids (not shown), and Dragendorff reagent (Sigma) for choline (not shown)
DPG: diphosphatidylglycerol; PG: phosphatidylglycol; AL: (ii) Aminolipid; PL: phospholipidd; PE: phospathylletholomine; PGNL: phosphoaminoglyolide; f: a first dimension; s: a second dimension.
FIG. 3 is a diagram of a 16S rRNA-based phylogenetic tree constructed from Halovibrio sp.YL5-2 and Halovibrio dentificans DSM15503 of Vibrio salina.
This figure is used to describe the species Vibrio salina Halovibrio sp.YL5-2 and Halovibrio dentificas DSM15503 and others. Numerical representation confidence values at phylogenetic tree branch points>50% (1000 replicates). The scale indicates that 5 of the 100 nucleotides were replaced. Pontibacter mucosus PB3TAs a foreign group. Each strain is assigned the GenBank accession number.
FIG. 4 is a phylogenetic tree diagram constructed by Halovibri sp.YL5-2 and Halovibri variabilis DSM 3050T of Vibrio salina and other strains based on the maximum simple Method (MP) of 16S rRNA.
This figure is used to describe the relationship between Halovibri sp. YL5-2 and the taxonomic groups of Halovibri variabilis DSM 3050T and other species. Marinobacter outlenesis Set74T(AY130994) as an outer group. GenBank accession numbers are in parentheses. Numerical representation confidence values at phylogenetic tree branch points>50% (1000 replicates)
FIG. 5 is a phylogenetic tree diagram constructed by Halovibrio sp.YL5-2 of Vibrio halophilus based on GGD matrix.
The figure shows a genome-genome (GGD) -based phylogenetic tree constructed using software FastME, with the outer group Marinobacterium aestuarii ST58-10T as root node
Biological material preservation information
YL5-2, classified and named as Halovibro salipaludis, the preservation date is 20 days 8 months in 2018, the preservation unit is the common microorganism center of China Committee for culture Collection of microorganisms, the preservation address is the institute of microorganisms of China academy of sciences, No. 3, institute of China academy of sciences, Ministry of China, Navy, the preservation number is: CGMCC NO. 16315.
Detailed Description
Example 1 isolation and preservation of Halovibrio sp.YL5-2 Vibrio salina
Vibrio salinophilus Halovibro sp.YL5-2 is separated from deposited soil of gelmu kha salt lake (36 degrees 51 'N and 94 degrees 95' E) in Qinghai province of China. The water of the salt lake of Kerr is at or close to the saturated salt concentration all the year round.
Preparing an LB liquid culture medium with NaCl concentration of 20%, adding 250mg/L of glycerol, 250mg/L of glucose and 50mg/L of methanol, and carrying out enrichment culture on the soil deposited in the salt lake of Carlsan at the temperature of 30-35 ℃ for 24-72 h. And (4) separating the strains in the enrichment culture solution by using YL solid culture medium. 1L of the culture medium contains the following components: glucose: 0.6g, trisodium citrate 0.5g, glycerol 2mL, yeast extract 0.8g, peptone 1.6g, dipotassium hydrogen phosphate 0.35g, potassium dihydrogen phosphate 0.1g, ammonium sulfate 0.25g, ammonium chloride 0.25g, MgSO4 0.5g，CaCl20.1g and NaCl 180 g; the trace element SL-410 mL, the pH value is 7.0-7.2; agar 2.5%.
Strain YL5-2 was obtained by plate streaking and repeated streaking purifications, with smooth edges and pale yellow color of colonies. A transmission electron micrograph of YL5-2 cultured on YL medium at 37 ℃ for 24h is shown in FIG. 1.
The strain YL5-2 is preserved in China general microbiological culture collection center with the preservation number of CGMCC NO.16315 and the preservation date of 2018, 8 months and 20 days.
Example 2 DNA sequencing, 16S rRNA sequence analysis, DNA hybridization assay and ANI analysis and identification of Halovibrio salina sp.YL5-2
The Genomic DNA of the strain YL5-2 was extracted by using TaKaRa Kit (TaKaRa MiniBEST bacterial Genomic DNA Extraction 68Kit Ver.3.0).
16S rRNA was amplified using the universal bacterial primers 27F (5-AGAGTTTGATCMTGGCTCA G-3) and 1492R (5-TACGGYTACCTTGTTACGACTT-3). Sequencing cycle conditions were: the initial temperature is maintained at 95 deg.C for 7min, and then circulation is carried out 35 times, each cycle comprises denaturation at 95 deg.C/0.5 min, annealing at 55 deg.C/0.5 min, extension at 72 deg.C/1.5 min, and final extension at 72 deg.C for 7 min. Purification of PCR products was performed using a Purification Kit (MinElute PCR Purification Kit, QIAGEN). PCR sequencing was performed by Shanghai Biotechnology Ltd. The complete 16S rRNA sequence of the strain YL5-2 is 1518bp, and the GenBank accession number is MF 782425.
The 16S rRNA sequence of strain YL5-2 was submitted to NCBI GenBank for sequence alignment (http:// www.ncbi.nlm.nih.gov /). And respectively adopting three methods, namely an NJ adjacency method, an MP maximum reduction method and an ML maximum likelihood method, constructing a phylogenetic tree in MEGA7 software, and repeatedly testing the reliability of the result of the topological developmental tree by 1000 times. The 16S rRNA-based phylogenetic tree showed that strain YL5-2 belongs to the genus Halovibrio (FIGS. 3,4 and 5), and was found to be related to Halovibrio variabilis DSM3050TMost closely related, among them the strain YL5-2 was associated with Halovibro variabilis DSM3050THas a similarity of 97.7% to Halovibro dentificas DSM15503TThe similarity with Halospina dentificas HGD 1-3 was 97.3%, and the similarity with Halospina dentificas HGD 1-3 was 95.5%. The similarity level of the 16S rRNA gene sequence between the strain YL5-2 and other strains in Halospina is lower than 97.0 percent, which shows that the low homology of the strain YL5-2 and the similar genome in GenBank shows that the strain YL5-2 is a new species which is different from other strains in Halovibrio.
The whole genome sequencing of the strain YL5-2 adopts a high-throughput sequencing platform of Illumina MiSeq 2000, Shanghai Paixino Biotech Co., Ltd. Raw sequencing data were filtered and corrected using prinsoq (version number v 0.20.4) software, followed by base pairing of the genome using SOAP denovo software (version number v1.05) with default parameters, and then evaluation of genome integrity using CheckM software (version 1.03). Protein-encoded open reading frames were predicted using Glimmer software (version number 1.2). RNA prediction using RNAmmer software (version 1.2). The total genome sequence of the strain YL5-2 is 3,495,096bp, and the GenBank accession number is NSKD 00000000.1.
DNA-DNA hybridization experiments were carried out with strain YL5-2 and the most genetically developed strain of the Halovibrio model of the genus Vibrio. The method is provided by De Ley equal to 1970, and DNA hybridization value (dDDH) is obtained by one-to-one comparison of gene sequences by adopting a 2 nd mode (version number 2.0) of GGDC software. The results of DNA tests and analyses showed that the strain YL5-2 and Halovibro varilabilis DSM3050T、Halovibrio denitrificans DSM15503TThe DNA-DNA hybridization values of (1) were 43.5% and38.2%, well below the 70% threshold (a generally accepted threshold for speciation).
The nucleotide average identity value was obtained by 1000 repetitive topological tests using the base set of the whole genome sequence. The method is proposed by Goris equal to 2007, and the software used is MUMmer (version number 3.23) and Jspecies (version number 1.2.1). Based on the ANI threshold range (95-96%) of the species division proposed by Kim et al and Richter et al, ANI analysis was performed on the genome of strain YL5-2 and on its closely related genomes in GenBank (Table 1). The result showed that the Average Nucleotide Identity (ANI) value of the strain YL5-2 with Tamilnaduibacter alinus Mi 7 was the highest and 88.5%, which is the strain YL5-2TA new species belonging to the genus Halovibrio provides argumentation.
TABLE 1 mean nucleotide identity (ANI) and DDH values between Halovibrio sp. YL5-2 of Vibrio salina and closely related genomes in GenBank.
Figure GDA0003307305600000071
Example 3 phenotypic characterization and physiological and biochemical characterization of Vibrio halophilus Halovibrio sp.YL5-2
Halovibrio salina sp. YL5-2 and another 2 model bacteria of the genus Vibrio, Halovibrio dentitilians DSM15503 and Halovibrio variabilis DSM3050, were phenotypically and biochemically identified at the same time.
Gram staining characteristics were tested using the BD gram staining kit.
Cell motility was determined using semi-MA medium (0.5% agar, w/v).
Cell morphology was examined by Transmission Electron Microscopy (TEM) analysis. That is, cells were picked from the exponentially growing culture solution, stained with 0.5% uranyl acetate, and photographed under a microscope (Tecnai Spirit, FEI, Hillsboro, OR, USA).
Oxidase activity Using oxidase kit (bioMerieux), by adding 3.0% H2O2The solution was poured into bacterial colonies and observed for bubble generation to determine catalase activity.
Temperature growth conditions were performed on YL liquid agar medium at 4, 10, 15, 20, 25, 30, 33, 37, 40, 45 and 50 ℃ respectively, with a constant pH of 7.5, comparing the strain YL5-2 at different temperaturesTThe growth rate determines its optimum growth temperature.
Salt tolerance was performed in YL agar and YL broth with 0.0-30.0% NaCl (w/v). Using buffer (Na)2HPO4/NaH2PO4(pH 5.0-7.0)，Na2CO3/NaHCO3(pH 8.0-12.0)) the pH was adjusted to 5.0, 5.5, 6.0, 7.0, 8.0, 9.0, 10.0 and 11.0 (15.0% NaCl, 35 ℃) to determine the pH range suitable for growth.
Carbon source utilization capacity and enzyme activity assays API 20NE, API ZYM (bioMerieux) and Biolog GENIII microplates were used. All tests inoculated cells pre-grown on YL medium and diluted with relevant inoculation medium.
The phenotypic and biochemical characterization results of strain YL5-2 are shown in Table 2.
TABLE 2 Halovibrio salina Halovibrio sp. YL5-2(a) and Halovibrio dentificas DSM15503T(b)，Halovibrio variabilis DSM 3050T(c) Comparison of phenotypic and physiobiochemical distinguishing characteristics
Figure GDA0003307305600000081
Description of the drawings: positive; negative.
Halovibrio sp. YL5-2 of Vibrio salina is gram-negative, aerobic, straight rod-like or Vibrio mimicus shape of 0.5-0.8X1.0-3.5 μm and moves through unipolar flagella (FIG. 1). The strain YL5-2 was grown aerobically with acetic acid and anaerobically with nitrate (API 20 NE). The results of the Biolog GENIII microplate test also showed that YL5-2 clearly distinguished the physiological and biochemical characteristics of Halovibro variabilis DSM3050 and Halovibro dentifices DSM15503 and other relatives.
Example 4 identification of fatty acids in Halovibrio sp.YL5-2 cells of Vibrio salina
Identification of cellular fatty acids YL5-2, Halovibro denitificans DSM15503 and Halovibro variabilis DSM3050 cells were cultured on YL medium at 30 ℃ for 3 days. The method mainly comprises the following steps: scraping 100mg of cells from YL medium and saponifying with 50% methanol containing sodium hydroxide; the saponified cells were freeze-dried and then extracted for cellular fatty acids using chloroform/methanol/0.3% (w/v) NaCl aqueous solution at a ratio of 1:2:0.8 (v/v/v).
The total amount of cellular fatty acids was determined using the phosphomolybdic acid method.
Fatty acid qualitative and quantitative determinations were performed using a 6890N gas chromatograph (Agilent) and standard MIS library generation software in the Sherlock microbiological identification system (VERSION 6.0and Date 4, Microbial ID Inc., Newark, DE, USA) (Sasser, 1990).
Cell fatty acid-based identification of species was performed using DSMZ-related analytical tools in germany.
The results of cellular fatty acid identification are shown in FIG. 2.
The only respiratory quinone of YL5-2 was ubiquinone Q-9, identical to Halovibro variabilis DSM 3050.
The major cellular fatty acids of YL5-2 include C18:1ω9c、C16:0、C19:0cyclo ω 8c and Summed feed 8 (table 3); the major polar lipids are diphosphatidylglycerol (dpg), phosphatidylethanolamines (pe), phosphatidylglycerol (pg), phosphatidyllcholine (pc) and two unidentified lipids (L). These are similar to the generic species Halovibrio varilabiliss and Halovibrio dentificas of YL 5-2.
TABLE 3 Halovibrio salina sp. YL5-2(a) and Halovibrio denitificans DSM15503T(b)，Halovibrio variabilis DSM 3050TComparison of cell fatty acid composition (%).
Figure GDA0003307305600000091
Figure GDA0003307305600000101
Description of the drawings:*summed diets denote a mixture of two or three fatty acids that cannot be separated by GLC from the MIDI system. The Summed feature 3 includes C16:1Omega 7C and/or C16:1Omega 6C, Summed feature 8 includes C18:1Omega 6C and/or C18:1ω7c。
Example 5 fermentation culture test of Halovibrio sp.YL5-2 of Vibrio salina
(1) The components of the culture medium are 500mg/L of glycerol, 250mg/L of glucose, 500mg/L of methanol, 200mg/L of methylamine, 100-250 g/L of sodium chloride, 250mg/L of sodium acetate, 250mg/L of trisodium citrate, 100mg/L of yeast powder, 200mg/L of peptone and 200mg/L of beef extract, a small amount of trace elements and the pH value of 7.5-8.0. And sterilizing the liquid culture medium, and simultaneously controlling the water evaporation.
(2) After being inoculated in a 1L triangular flask, the culture is carried out for 48 hours at the temperature of 35 ℃, and the change of the salt concentration is influenced by the dissipation of the supplemented water in the culture process. After 48h, the OD600 values at 10%, 15%, 20% and 25% salt concentration were 1.72, 1.65, 1.62, 1.60 and 1.63, respectively. And performing two times of transfer culture, wherein the culture is performed for 48 hours after each transfer, and the OD600 is respectively 2.56, 2.68, 2.72, 2.68 and 2.52 after the acclimatization culture is completed.
(3)1L of the culture broth was inoculated into a 20L aerobic fermentor and cultured, with the medium composition remaining unchanged. Still culturing for 48h at 35 deg.C under stirring speed of 100rpm with dissolved oxygen of 2.0-4.0 mg/L. After 48 hours, the OD600 of the bacterial liquid in the fermentation tank can reach 2.6-3.0.
(4) And (4) checking: sampling and observing every day by using a microscope in the culture process, and checking whether mixed bacteria grow; and meanwhile, the growth and morphological change conditions of YL5-2 are observed.
(5) As a result: test results show that the strain YL5-2 can be rapidly subjected to fermentation culture and amplification, which shows that YL5-2 has the potential of large-scale engineering application.
Example 6 salt degradation resistance test of Halovibrio sp.YL5-2 Vibrio halophilus
(1) The components of the culture medium are 50mg/L of glycerol, 25mg/L of glucose, 50mg/L of methanol, 20mg/L of methylamine, 250g/L of sodium chloride, 25mg/L of sodium acetate, 25mg/L of trisodium citrate, 10mg/L of yeast powder, 20mg/L of peptone, 20mg/L of beef extract and 20g/L of agar.
(2) Activating strains with fresh culture medium, and culturing for 3 days until thallus Porphyrae grows abundantly
(3) Salinity gradient setting: according to the enrichment screening conditions of YL5-2 strain, the salinity gradient and range of the culture medium are set to be 0%, 0.5%, 1%, 2%, 3%, 24%, 26%, 28%, 30% and 32%.
(4) Preparing a culture medium: the culture medium is prepared according to the set culture medium formula, the culture medium with higher salinity is sterilized after being melted by hot water, 5ml of evaporated water is added in each bottle, the volatile culture medium components are added after being sterilized and are evenly shaken, the plate is poured after being cooled to about 60 ℃, and the salinity is high and is easy to solidify, so that the pouring of the plate is fast (when the culture medium with 32% salinity is poured into the plate, a small amount of salt is precipitated after cooling and solidification, and a large amount of salt crystals are precipitated at 34% salinity).
(5) Inoculation: selecting a ring of fresh lawn under aseptic condition, inoculating to culture medium with various salinity, transferring from low salinity to high salinity in gradient manner, streaking with 34% salinity culture medium, separating out a large amount of salt crystals along with streaking track, sealing the culture dish with sealing film after inoculation, culturing at 35-37 deg.C for 3-7 days, and observing strain growth condition.
(6) And (3) test results: the salt tolerance range of YL5-2 strain is 3-32%. The YL5-2 strain can obviously observe newly grown lawn in 3-30% salinity culture medium within three days, but can grow to naked eyes in 32% saturated salinity culture medium for more than 7 days. The growth rate of YL5-2 is higher in the salt concentration range of 3% -30%.
Example 7 microbial Agents prepared Using Vibrio halophilus Halovibrio sp.YL5-2
(1) The components of the culture medium are 500mg/L of glycerol, 250mg/L of glucose, 500mg/L of methanol, 200mg/L of methylamine, 100-250 g/L of sodium chloride, 250mg/L of sodium acetate, 250mg/L of trisodium citrate, 100mg/L of yeast powder, 200mg/L of peptone and 200mg/L of beef extract, a small amount of trace elements and the pH value of 7.5-8.0. And sterilizing the liquid culture medium, and simultaneously controlling the water evaporation.
(2) Inoculating Halovibrio halophilus Halovibrio sp. YL5-2 into a 1L triangular flask, culturing at 35 deg.C for 48h, and supplementing water during culture to affect change of salt concentration. After 48h, the OD600 values at 10%, 15%, 20% and 25% salt concentration were 1.72, 1.65, 1.62, 1.60 and 1.63, respectively. And performing two times of transfer culture, wherein the culture is performed for 48 hours after each transfer, and the OD600 is respectively 2.56, 2.68, 2.72, 2.68 and 2.52 after the acclimatization culture is completed.
(3)1L of the culture broth was inoculated into a 20L aerobic fermentor and cultured, with the medium composition remaining unchanged. Still culturing for 48h at 35 deg.C under stirring speed of 100rpm with dissolved oxygen of 2.0-4.0 mg/L. After 48 hours, the OD600 of the bacterial liquid in the fermentation tank can reach 2.6-3.0.
(4)20L of the culture broth was inoculated into a 1000L aerobic fermentor and cultured under the same conditions as described above. The culture can then be scaled up or cultured in bulk as desired.
(5) The obtained bacterial liquid is the halophilic vibrio haloovibro sp.YL5-2 microbial agent.
(6) The microbial inoculum is used for treating wastewater with TDS of 3% and acetic acid of 1000mg/L, and the degradation efficiency of the acetic acid exceeds 90% after 72 hours.
(7) When the microbial inoculum is used for treating wastewater with TDS of 6% and acetic acid of 1000mg/L, the acetic acid degradation rate can reach 90% in 48h and can reach more than 95% in 72 h.
(8) The microbial inoculum is used for treating wastewater with TDS of 10% and acetic acid of 1000mg/L, and after 48 hours, the acetic acid degradation rate can reach more than 95%.
(9) The microbial inoculum is used for treating wastewater with TDS of 15% and acetic acid of 1000mg/L, and after 48 hours, the acetic acid degradation rate can reach more than 95%.
(10) The microbial inoculum is used for treating wastewater with TDS of 20% and acetic acid of 1000mg/L, and after 48, the acetic acid degradation rate can reach more than 95%.
(11) The microbial inoculum is used for treating wastewater with TDS of 25% and acetic acid of 1000mg/L, and after 48 hours, the acetic acid degradation rate can reach more than 95%.
(12) The microbial inoculum is used for treating wastewater with TDS of 30% and acetic acid of 1000mg/L, and after 48 hours, the acetic acid degradation rate can reach more than 90%.
(13) The microbial inoculum is used for treating wastewater with TDS of 32% and acetic acid of 1000mg/L, and the acetic acid degradation rate can reach more than 90% after 72 hours.
(14) The microbial inoculum is used for treating wastewater with TDS of 34% and acetic acid of 1000mg/L, and after 72 hours, the acetic acid degradation rate can reach more than 60%; after 144 hours, the acetic acid degradation rate can reach more than 90%.
Example 8 composite microbial agent prepared by using Halovibrio sp.YL5-2 of Vibrio salina
(1) The components of the culture medium are 500mg/L of glycerol, 250mg/L of glucose, 500mg/L of methanol, 200mg/L of methylamine, 120g/L of sodium chloride, 250mg/L of sodium acetate, 250mg/L of trisodium citrate, 100mg/L of yeast powder, 200mg/L of peptone and 200mg/L of beef extract, a small amount of trace elements and the pH value of 7.5-8.0. And sterilizing the liquid culture medium, and simultaneously controlling the water evaporation.
(2) A1L triangular flask is simultaneously inoculated with Halovibrio halophilic sp.YL5-2 and Halovibrio densitificans DSM15503, and then cultured for 48h at 35 ℃, and the change of salt concentration is influenced by the loss of supplemented water in the culture process. The transfer is carried out every 72h of culture, and the OD600 is not less than 1.5 before each transfer.
(3)1L of the culture broth was inoculated into a 20L aerobic fermentor and cultured, with the medium composition remaining unchanged. Still culturing for 48h at 35 deg.C under stirring speed of 100rpm with dissolved oxygen not less than 2.0 mg/L. After 48h, the OD600 of the bacterial liquid in the fermenter reached 3.0.
(4)20L of the culture broth was inoculated into a 1000L aerobic fermentor and cultured under the same conditions as described above. The culture can then be scaled up or cultured in bulk as desired.
(5) The obtained bacterial liquid is the compound microbial agent consisting of Halovibro sp.YL5-2 and Halovibro dentificas DSM 15503.
(6) The microbial inoculum is used for treating wastewater with TDS of 3%, 6%, 10%, 15%, 20%, 25% and 30% respectively, and acetic acid of 1000mg/L, and the degradation rates of the acetic acid after 72h are 90.3%, 92.6%, 92.7%, 92.8%, 91.8%, 90.7% and 89.5% respectively.
(7) The TDS of the microbial inoculum is respectively 3%, 6%, 10%, 15%, 20%, 25% and 30% when the microbial inoculum is treated under 35 ℃ and anoxic conditions, and the acetic acid is 1000mg/L, NO3the-N is less than or equal to 100mg/L, and the degradation rates of the acetic acid after 72 hours are respectively 84.5%, 91.2%, 91.8%, 92.3%, 92.2%, 88.5% and 83.2%.
(8) The bacteria agent is used for treating TDS at 30 ℃ and under the anoxic condition of 3%, 6%, 10%, 15%, 20%, 25% and 30% respectively, and acetic acid is 1000mg/L, NO2The acetic acid degradation rate after 72 hours of the wastewater of less than or equal to 100mg/L is 83.6 percent, 90.7 percent, 92.0 percent, 91.5 percent, 91.9 percent and 84.2 percent respectively.
(9) The microbial inoculum is used for treating TDS at 25 deg.C under anoxic condition of 3%, 6%, 10%, 15%, 20%, 25%, 30%, and COD is less than or equal to 3000mg/L, NO3Waste water of less than or equal to 100mg/L and NO after 72 hours3The degradation rates are respectively 88.3%, 93.1%, 95.5%, 95.6%, 92.8% and 86.6%.
Example 9 composite microbial agent prepared by using Halovibrio sp.YL5-2 of Vibrio salina
(1) The components of the culture medium are 500mg/L of glycerol, 250mg/L of glucose, 500mg/L of methanol, 200mg/L of methylamine, 200g/L of sodium chloride, 250mg/L of sodium acetate, 250mg/L of trisodium citrate, 100mg/L of yeast powder, 200mg/L of peptone and 200mg/L of beef extract, a small amount of trace elements and the pH value of 7.5-8.0. And sterilizing the liquid culture medium, and simultaneously controlling the water evaporation.
(2) Inoculating Halovibrio halophilus sp. YL5-2 into a 1L triangular flask, culturing at 35 deg.C for 48h, and supplementing water to affect the change of salt concentration. The transfer is carried out every 72h of culture, and the OD600 is not less than 1.5 before each transfer.
(3)1L of the culture broth was inoculated into a 20L aerobic fermentor and cultured, with the medium composition remaining unchanged. Still culturing for 48h at 35 deg.C under stirring speed of 100rpm with dissolved oxygen of not less than 2.0 mg/L. After 48h, the OD600 of the bacterial liquid in the fermenter reached 2.5.
(4)20L of the culture broth was inoculated into a 1000L aerobic fermentor and cultured under the same conditions as described above. The culture can then be scaled up or cultured in bulk as desired.
(5) And mixing the cultured bacterial liquid with a bacterial liquid obtained by enrichment culture of seawater under the condition that the TDS is 10%, wherein the obtained bacterial liquid is the compound microbial agent containing vibrio halophilus Halovibro sp.
(6) The microbial inoculum is used for treating the preparation wastewater with TDS of 10% and glucose of 1000mg/L, and the removal rate of the glucose can reach more than 98% after 72 hours; the microbial inoculum is applied to epoxypropane wastewater treatment, an activated sludge process and a contact oxidation process are adopted for continuous domestication for one month, and the COD removal rate of the whole process can be stabilized at 80-85% under the condition that the inlet water COD is 4-5% of 900-1200 mg/L, TDS; the microbial inoculum is applied to the treatment of preserved tuber mustard wastewater, and adopts the process of 'activated sludge process + flocculation precipitation' under the conditions that the TDS of the inlet water is less than or equal to 7 percent and the COD is less than or equal to 3000mg/L, and the average removal rate of the COD is more than 90 percent; the microbial inoculum is applied to the treatment of wastewater of epoxy chloropropane, and the average removal rate of TOC can reach more than 60% by adopting a contact oxidation process under the conditions that the TDS of inlet water is 21-23% and the TOC is less than or equal to 1000 mg/L.
(7) The composite microbial inoculum is applied to the soil polluted by high-salt pesticide wastewater, and test results show that after the composite microbial inoculum is added, the COD removal rate of a soil leaching solution is increased by about 30% compared with the COD removal rate of a control group without the composite microbial inoculum.
(8) The composite microbial inoculum is applied to ecological restoration of the sea mouth polluted wetland.
(9) The composite microbial inoculum is applied to remediation of polluted groundwater in offshore areas, and the degradation speed of COD in the polluted groundwater is obviously accelerated by adding the microbial inoculum and hydraulic circulation.
Example 10 composite microbial agent prepared by using Halovibrio sp.YL5-2 of Vibrio salina
(1) The components of the culture medium are 500mg/L of glycerol, 250mg/L of glucose, 500mg/L of methanol, 200mg/L of methylamine, 200g/L of sodium chloride, 250mg/L of sodium acetate, 250mg/L of trisodium citrate, 100mg/L of yeast powder, 200mg/L of peptone and 200mg/L of beef extract, a small amount of trace elements and the pH value of 7.5-8.0. And sterilizing the liquid culture medium, and simultaneously controlling the water evaporation.
(2) Inoculating Halovibrio halophilus sp. YL5-2 into a 1L triangular flask, culturing at 35 deg.C for 48h, and supplementing water to affect the change of salt concentration. The transfer is carried out every 72h of culture, and the OD600 is not less than 1.5 before each transfer.
(3)1L of the culture broth was inoculated into a 20L aerobic fermentor and cultured, with the medium composition remaining unchanged. Still culturing for 48h at 35 deg.C under stirring speed of 100rpm with dissolved oxygen not less than 2.0 mg/L. After 48 hours, the OD600 of the bacterial liquid in the fermentation tank reaches more than 3.0.
(4)20L of the culture broth was inoculated into a 1000L aerobic fermentor and cultured under the same conditions as described above. The culture can then be scaled up or cultured in bulk as desired.
(5) And mixing the cultured bacterial liquid with a bacterial liquid obtained by enrichment culture of seawater under the condition that TDS is more than 10 percent to obtain the compound microbial agent containing vibrio halophilus Halovibrio sp.YL5-2.
(6) The aerobic process is adopted to verify the treatment effect of the microbial inoculum on the high-salinity wastewater, and the test result is as follows.
Figure GDA0003307305600000151

Claims (1)

1. The application of the compound microbial agent in wastewater treatment and soil remediation; the wastewater is selected from propylene oxide wastewater, preserved szechuan pickle wastewater, epichlorohydrin wastewater and polluted underground water in offshore areas; the soil is selected from the soil polluted by high-salt pesticide wastewater; the compound bacterial agent is Vibrio halophilus with preservation number CGMCC NO.16315Halovibrio salipaludis Mixing YL5-2 bacterial solution and bacterial solution obtained by enrichment culture of seawater under TDS > 10%.
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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103074286A (en) * 2013-01-28 2013-05-01 北京大学 High-salt heterotrophic nitrification-aerobic denitrification dephosphorization salinivibrio and application of salinivibrio in wastewater treatment
CN105886444A (en) * 2016-06-25 2016-08-24 山东文远建材科技股份有限公司 Compound microbial agent as well as preparation method and use method thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103074286A (en) * 2013-01-28 2013-05-01 北京大学 High-salt heterotrophic nitrification-aerobic denitrification dephosphorization salinivibrio and application of salinivibrio in wastewater treatment
CN105886444A (en) * 2016-06-25 2016-08-24 山东文远建材科技股份有限公司 Compound microbial agent as well as preparation method and use method thereof

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
Extremely halophilic denitrifying bacteria from hypersaline inland lakes, Halovibrio denitrificans sp. nov. and Halospina denitrificans gen. nov., sp. nov., and evidence that the genus name Halovibrio Fendrich 1989 with the type species Halovibrio;D Yu Sorokin等;《Int J Syst Evol Microbiol》;20060228;第56卷(第2期);379-388 *
Halovibrio sp. strain YL5-2 16S ribosomal RNA gene, partial sequence;Zhang X 等;《Genbank登录号：MF782425》;20170914;1 *

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