CN117535166B - Special acid-feeding dyford bacterium for ensiling of bromhidrosis and application of special acid-feeding dyford bacterium - Google Patents
Special acid-feeding dyford bacterium for ensiling of bromhidrosis and application of special acid-feeding dyford bacterium Download PDFInfo
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- CN117535166B CN117535166B CN202211694988.7A CN202211694988A CN117535166B CN 117535166 B CN117535166 B CN 117535166B CN 202211694988 A CN202211694988 A CN 202211694988A CN 117535166 B CN117535166 B CN 117535166B
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- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 4
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- 239000004310 lactic acid Substances 0.000 description 2
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- OAOABCKPVCUNKO-UHFFFAOYSA-N 8-methyl Nonanoic acid Chemical compound CC(C)CCCCCCC(O)=O OAOABCKPVCUNKO-UHFFFAOYSA-N 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K30/00—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs
- A23K30/10—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder
- A23K30/15—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder using chemicals or microorganisms for ensilaging
- A23K30/18—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder using chemicals or microorganisms for ensilaging using microorganisms or enzymes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Microbiology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Polymers & Plastics (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Food Science & Technology (AREA)
- Animal Husbandry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Fodder In General (AREA)
Abstract
The invention discloses a delford bacterium acidovorans and application thereof. The delford bacterium acidovorans is the delford bacterium acidovorans Delftia acidovorans; the preservation name is Thermomyces lanuginosus LL2022006; the microbial strain is preserved in China general microbiological culture Collection center (CGMCC), and the preservation address is 1 # 3 of North West Lu in the Chaoyang area of Beijing city; preservation date: 2022, 4, 7; the preservation number is CGMCC NO.24652. The 16S rDNA gene sequence of the acid-feeding dyford bacteria is a nucleotide sequence shown as SEQ ID No. 1. The invention can inhibit the growth of staphylococcus aureus, salmonella typhimurium and escherichia coli K88, has strong bacteriostasis and acidity, and can improve the silage quality of the foxtail algae.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to acid-feeding dyford bacteria special for ensiling of bromhidrosis and application thereof.
Background
Lactic acid bacteria are one of the common microbial additives for feed in the livestock and poultry breeding process. It can ferment to produce acid, reduce pH value of environment and prolong shelf life. Research shows that lactic acid bacteria can promote animal growth, immunity and disease resistance and raise silage quality of green crops.
The foxtail is a green feed resource applied to water quality purification, is commonly used for absorbing nitrogen pollutants in water bodies, and achieves the effect of purifying water quality. However, because of two limiting factors such as contact with sewage and higher water content, the foxtail is easy to rot after harvesting, and how to better apply the foxtail as a green feed resource, so that the secondary pollution is avoided.
The livestock and poultry industry development process in China faces the problems that feed is in shortage, and protein and energy feed resources cannot be self-sufficient; at present, the consumption of raw grains for feed in China is accumulated to be more than 40% of the total consumption of all grains in China, the proportion of the raw grains for feed is more than 50%, the problem that feed resources are short enough and people and livestock contend for the grains is more remarkable, and the problem that the feed resources are short enough in winter for herbivores and even contend with the human beings is solved, so that the novel unconventional feed resources are developed to be an effective way for relieving the hidden depletion of the feed resources and promoting and guaranteeing the healthy and sustainable development of animal husbandry while the existing feed resources are better utilized. The acid-feeding dyford bacteria has good acid production and bacteriostasis properties, has the property of inhibiting pathogenic bacteria and promoting growth, and can be used as a novel feed additive for fermenting novel pasture.
At present, there is a lack of special acid-feeding dyford bacteria for ensiling the foxtail algae with strong bacteria inhibition and application thereof.
Disclosure of Invention
The invention aims to solve the technical problem of providing the special acid-feeding dyford bacterium for ensiling the gulfweed with strong bacteriostasis and the application thereof.
In order to achieve the above purpose, the present invention is realized by the following technical scheme: the invention relates to a special acid-feeding dyford bacterial strain for ensiling the bromhidrosisDelftia acidovorans) The preservation name of the special delfurd bacterium strain for ensiling the bromhidrosis is delfurd bacterium LL2022006; the preservation unit is China general microbiological culture Collection center (CGMCC), and the preservation address is North Xishlu No.1 and No. 3 in the Korean area of Beijing; preservation date: 2022, 4, 7; the preservation number is CGMCC NO.24652.
Further, the 16S rDNA gene sequence of the special acid-feeding dyford bacterial strain for ensiling the foxtail algae is a nucleotide sequence shown as SEQ ID No. 1.
Further, the colony morphology and gram staining microscopy thereof are characterized by: in MRS agar culture medium, after aerobic culture for 72 hours at 35 ℃, bacterial colony is milky round, smooth in surface, neat in edge and easy to pick up; gram staining is positive, and the thalli are rod-shaped, single or double, and are facultative anaerobic.
The invention relates to a special acid-feeding dyford microbial inoculum for ensiling the bromhidrosis.
Further, the active ingredients are at least one of the following (a), (b) and (c):
(a) The fermentation culture of the special acid-feeding delfurd microbial inoculum for the ensiling of the foxtail algae;
(b) The obtained spore suspension of the special acid-feeding delforskolin microbial agent for the ensiling of the foxtail algae;
(c) The obtained special acid-feeding delfurd bacterial agent cells for the ensilage of the foxtail algae are subjected to ultrasonic pyrolysis and precipitation.
The preparation method of the special acid-feeding delfurd microbial inoculum for ensiling the foxtail algae comprises the following steps: (1) strain preparation: inoculating v=0.1% in MRS broth culture medium, culturing at 35-37 ℃ for 36~48 h,5000~8000 rpm, and centrifuging to obtain thalli;
(2) Freeze-drying the thalli to prepare the special acid-feeding dyford fungus preparation for ensiling the foxtail algae.
Further, in step (1), the MRS medium broth is: peptone 10.0; beef extract 5.0; 4.0 parts of yeast powder; glucose 20.0; dipotassium hydrogen phosphate 2.0; 2.0 parts of citric acid triammonium; 5.0 parts of sodium acetate; magnesium sulfate 0.2, manganese sulfate 0.05, tween 80 1.0mL.
Further, in the step (2), the effective viable count of the cells in the microbial preparation is 10 10 ~10 11 CFU/g。
The invention relates to the use of the Thermomyces acidovorus or Thermomyces acidovorus in feed (including turquoise feed).
The delfurd acid-eating bacteria or the delfurd acid-eating bacteria agent can be applied to water, therapeutic drugs or health care drugs.
The beneficial effects are that: the invention has strong bacteriostasis and acid production, and can improve the silage quality of the foxtail algae.
Compared with the prior art, the invention has the following advantages:
the acid-feeding dyford bacteria has remarkable antagonism on pathogenic bacteria, and can inhibit the growth of staphylococcus aureus, salmonella typhimurium and escherichia coli K88. The acid-feeding dyford bacteria provided by the invention can grow rapidly, reduce the pH and resist low temperature.
The invention provides the acid-feeding dyford bacteriaDelftia acidovorans) Isolated from naturally fermented pennisetum hydridum, and identified as Thermomyces acidovorus by colony morphology observation, gram staining, physiological and biochemical identification, molecular biology and the likeDelftia acidovorans) The strain has high growth speed and high acid production efficiency, and can be used for treating Escherichia coli K88 and golden yellowBoth staphylococcus and salmonella typhimurium have good inhibition.
(3) According to the fermentation experiment of the invention, the addition amount of 200ml of bacterial liquid is added into each ton of pasture, so that the odor score of the pasture can be improved, the pH value of the pasture can be reduced, the types of microorganisms in the fermented product are improved, compared with the comparison group, the bacterial types are improved by 32, after the patent bacteria are added, the effects of anti-inflammatory, mildew-proof and the like are improved, the production of high-quality short-chain fatty acid is promoted, the immunity function can be enhanced by improving the NK cell activity, gastric juice and bile salt can be tolerated, the cholesterol level can be reduced, and the relative abundance of Weishi genus with the effects of flavor and the like can be improved; reduction of rot in turquoise plantsBulleraThe relative abundance of the mold; the content of serine lysine with defensive action in the fermented product can be improved, and the content of antioxidant substance blue Mei Chun C glycoside can be improved; the reduction of the content of butyric acid, which is positively correlated with the loss of dry matter, results in an increase of soluble proteins and ammonium nitrogen and the content of butyric acid is inversely proportional to the quality of silage, and the increase of the content of isocapric acid, which is directly proportional to the digestibility of ruminants such as cows.
Drawings
FIG. 1 is a colony morphology of LL2022006 according to the present invention;
FIG. 2 is a view of a 1000 Xmicroscope of the present invention;
FIG. 3 is a diagram showing the judgment of sensory evaluation index of silage quality evaluation Standard according to the present invention.
Detailed Description
The invention is further described with reference to the drawings and specific embodiments. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention.
Example 1
The invention relates to a special acid-feeding dyford bacterial strain for ensiling the bromhidrosisDelftia acidovorans) The preservation name of the special delfurd bacterium strain for ensiling the bromhidrosis is delfurd bacterium LL2022006; the preservation unit is China general microbiological culture Collection center (CGMCC), and the preservation address is Beijing Chaoyang North Xicilu No.1Hospital number 3; preservation date: 2022, 4, 7; the preservation number is CGMCC NO.24652.
The 16S rDNA gene sequence of the acid-feeding dyford bacteria is a nucleotide sequence shown as SEQ ID No. 1.
In MRS agar culture medium, after aerobic culture for 72 hours at 35 ℃, bacterial colony is milky round, smooth in surface, neat in edge and easy to pick up; gram staining is positive, and the thalli are rod-shaped, single or double, and are facultative anaerobic.
The invention relates to a delfurd acid bacteria agent prepared by the delfurd acid bacteria.
The active ingredients are as follows:
(1) The fermentation culture of the acid-feeding dyford bacteria;
the preparation method of the acid-feeding delford microbial inoculum comprises the following steps:
(1) Preparing strains: inoculating in MRS liquid culture medium, culturing at 35 deg.C for 48 hr, centrifuging at 5000rpm to obtain thallus.
(2) Freeze-drying the cells.
In step (1), the MRS medium is: 10.0g of peptone; beef extract 5.0g; 4.0g of yeast powder; glucose 20.0g; 2.0g of dipotassium hydrogen phosphate; 2.0g of tri-ammonium citrate; 5.0g of sodium acetate; 0.2g of magnesium sulfate; manganese sulfate 0.05g; tween 80.0 ml; 1L of water.
The effective viable count of the bacterial cells in the microbial preparation is 1 multiplied by 10 11 CFU/g。
The invention relates to the use of the acid-feeding dyford bacteria or the acid-feeding dyford bacteria agent in feed and water.
Example 2
Example 2 differs from example 1 in that:
in MRS agar culture medium, after aerobic culture for 72 hours at 35 ℃, bacterial colony is milky round, smooth in surface, neat in edge and easy to pick up; gram staining is positive, thalli are rod-shaped, single or double bacteria are present, facultative anaerobism is carried out, and the optimal growth temperature is 35-37 ℃.
The preparation method of the acid-feeding delford microbial inoculum comprises the following steps:
(1) Preparing strains: inoculating MRS liquid culture medium, and culturing at 35deg.C for 48h to obtain bacterial liquid.
(2) The cells were stored at 4 ℃.
The effective viable count of the thalli in the microbial preparation is 10 9 CFU/ml。
Example 3
Example 3 differs from example 1 in that:
in MRS agar culture medium, after aerobic culture for 36h at 37 ℃, bacterial colony is milky round, smooth in surface, neat in edge and easy to pick up; gram staining is positive, thalli are rod-shaped, single or double bacteria are present, facultative anaerobism is carried out, and the optimal growth temperature is 35-37 ℃.
The preparation method of the intestinal membrane-like Weissella agent comprises the following steps:
(1) Preparing strains: inoculating in MRS liquid culture medium, culturing at 35deg.C for 48h, and centrifuging at 6000 rpm to obtain thallus.
(2) Freeze-drying the cells.
The effective viable count of the thalli in the microbial preparation is 1011CFU/g.
Test example 1
The acid-feeding dyford bacteria provided by the invention have the following microbial characteristics: in MRS agar culture medium, after aerobic culture for 72 hours at 35 ℃, bacterial colony is milky round, smooth in surface, neat in edge and easy to pick up; gram staining is positive, thalli are rod-shaped, single or double bacteria are present, facultative anaerobism is carried out, and the optimal growth temperature is 35-37 ℃.
Antibacterial property measurement:
the method comprises the steps of preparing suspension bacteria liquid by using escherichia coli K88, salmonella typhimurium and staphylococcus aureus as indicator bacteria and using 0.9% saline to adjust the concentration, preparing an agar plate containing about 105 CFU/mL of indicator bacteria, cooling, punching a pore diameter of 4 mm on the plate by a gel punching method, adding 60uL of bacteria liquid to be detected, and culturing at 35 ℃ for 17 hours to determine the diameter of a bacteria inhibition zone, wherein the results are shown in table 1. The in vitro antagonistic effect of LL2022006 on three pathogenic bacteria is shown in table 1:
table 1 antagonistic properties against pathogenic bacteria unit: mm (mm)
| Strain numbering | Coli K88 | Salmonella typhimurium | Staphylococcus aureus |
| LL2022006 | 13.97 | 17.80 | 12.57 |
Selecting the foxtail algae with the growing period of about 4 months, harvesting and cutting into sections (3-5 cm). The rice bran powder was used for moisture conditioning (rice bran powder was added at a rate of 33.3%) and randomly divided into 2 groups (1 test group and 1 control group) of 6 replicates each of about 20Kg per packet of 3 packets. According to the proportion of adding 20g of freeze-dried powder into each ton of forage grass (1000 ml of suspension is prepared from 20g of freeze-dried powder), the test period is 2022, 5, 20, 6, 16 and 27d. The same volume of water was added to the control group, the additive prepared in example 1 (suspension prepared in proportion) was added to the test group, the fermentation was carried out at room temperature, the test period was 35 days, and the silage effect was evaluated at the end of the test. The effect of addition of LL2022006 on the silage effect of gulfweed is shown in table 2:
table 2 in situ evaluation score
| pH | Smell of | Color | Structure of the | |
| Test group | 4.80 | 10.00 | 1.00 | 4.00 |
| Control group | 5.62 | 3.33 | 1.00 | 4.00 |
Test example 2
Test analysis
OTU (Operational Taxonomic Units) is a unified flag set for a person to a certain classification unit (strain, genus, species, group, etc.). The analysis of the wien diagram of fig. 3 shows that the number of OTUs in the fermentation product (MC) of the foxtail algae without adding the test strain LL2022006 is 199, the number of unique OTUs is 44, the number of OTUs in the test group (M1) with the dry bacterial powder added is 231, the number of unique OTUs is 76, and table 4 shows that the number of bacterial species in the control group is 32 less than that in the test group on a belonging level, so that the dry bacterial powder additive of the invention can extract the number of species in the foxtail algae fermentation product. The effect on intestinal flora structure at the level of fermentation of the gulfweed after addition of LL2022006 is shown in table 4:
TABLE 4% by weight
| Genus | Test group | Control group |
| Lactobacillus genusLactobacillus | 62.86±4.26 | 57.88±11.12 |
| Chloroplast norank_f __ norank_o __ chlorplast | 19.28 a ±4.7 | 35.82 b ±9.71 |
| Unclassified Enterobacteriaceae unclassified_f __ Enterobacteriaceae | 6.56 B ±2.28 | 2.21 A ±1.06 |
| Rhizobium dissimilaris (L.) DCAllorhizobium-Neorhizobium-Pararhizobium-Rhizobium | 1.33±0.51 | 0.89±0.17 |
| Weissella genusWeissella | 0.87 b ±0.34 | 0.37 a ±0.13 |
| Lactococcus genusLactococcus | 0.17±0.04 | 0.15±0.1 |
| Sphingomonas spNovosphingobium | 0.30±0.14 | 0.32±0.15 |
| Sphingomonas spNovosphingobium | 0.30±0.14 | 0.32±0.15 |
Note that: the same row of data shoulders indicates that the difference is significant (P < 0.05) and the difference is extremely significant (P < 0.01). The table below is the same.
As can be seen from Table 4, after addition of the proprietary strains, the unclassified Enterobacter and Weissella in the test group were significantly increased relative to the abundance and chloroplast were significantly decreased. The Weissella has remarkable anti-inflammatory and mildew-proof effects, promotes the production of high-quality short-chain fatty acid, can enhance the immune function by improving the activity of NK cells, can resist gastric juice and bile salts, has moderate cell surface hydrophobicity, reduces cholesterol level and improves flavor. Therefore, the addition of the strain has the effect of improving the quality of the fermentation product of the bromhidrosis. The effect on fungal flora structure at the level of the silage of gulfweed after addition of LL2022006 is shown in table 5:
as is clear from Table 5, after the addition of the patent bacteria, the relative abundance of the genus Brevibacterium, bullera and Glehniae was significantly decreased, and the genus Leptoradix was significantly increased. Bullera is positively related to the rot of green plants, so that the relative abundance of rot-related moulds can be reduced after the patent bacteria are added.
TABLE 5
| Test group | Control group | |
| Unclassified Rogowoad phylum unclassified_p __ Rozellomyceta | 5.22±2.91 | 5.45±1.79 |
| Cladosporium genusCladosporium | 13.04±2.51 | 11.28±2.55 |
| Powdery mildew genusErysiphe | 6.47±1.29 | 7.79±1.00 |
| Unclassified bipedaceae unclassified_f __ bipodascace | 0.03±0.02 | 0.03±0.02 |
| Saitozyma | 7.41±2.11 | 10.64±4.03 |
| Genus BrettanomycesFilobasidium | 2.86 a ±1.21 | 7.66 b ±3.22 |
| Aspergillus genusAspergillus | 2.96±1.52 | 3.95±1.82 |
| Vicina NikkeraVishniacozyma | 4.36±1.92 | 8.55±4.26 |
| Leptosporium (Leptosporum)Ampelomyces | 1.97±0.81 | 4.07±2.18 |
| Genus CoralliumHoltermanniella | 2.27 a ±0.88 | 5.23 b ±1.74 |
| Bullera | 2.72 a ±0.75 | 4.54 a ±1.44 |
| Genus LeptoradixEntyloma | 5.21 b ±3.25 | 1.59 a ±0.59 |
| Lock throw yeastSporidiobolus | 1.93±0.86 | 2.63±0.83 |
| Genus ArthrobacterWallemia | 1.92±0.47 | 2.04±0.84 |
| Alternaria genusAlternaria | 1.03±0.54 | 2.11±1.15 |
As can be seen from Table 6, the analysis of the metabolites in the fermentate, after finding the different metabolites, found: DG (20:1 (11Z)/15:0/0:0).
1- [ (2-chloropheny) methyl ] -N-cyclopeptidylpiperidine-3-carboxamide, serine lysine and blue enol C glucoside all rise significantly or very significantly, and hydrozylmethylglucose, methyl isoamylene, neosilane, 13-hydroxy abscisic acid, 3'-hydroxy-3,4,5,4' -tetramethoxystilbene and 3-deoxy-D-glycero-D-galactose-2-non-gluconic acid drop significantly. Methyl isoamylene belongs to a class of organic compounds known as piperidines and is a primary metabolite.
New silane organic compounds pyrrolizidine. Pyrrolizidine is a compound containing pyrrolizidine which is a bicyclic system consisting of two fused pyrrolizidine rings sharing one nitrogen atom. Serine lysine, also known as dipeptide, is an organic compound containing two alpha-amino acid sequences joined by exactly one peptide bond. Serine is a secondary metabolite. The secondary metabolite is a metabolic or physiological nonessential metabolite that can act as a defensive or signaling molecule. Lan Meichun C glycoside belongs to a class of sesquiterpene organic compounds, called fatty acyl glycosides of mono-and disaccharides, having antioxidant effect. Therefore, after the patent bacterium is added, the content of serine lysine with a defensive effect in the fermented product can be increased, and the content of antioxidant substance blue Mei Chun C glycoside can be increased.
TABLE 6
| Metabolite | FC(E/C) | P_value |
| Hydroxymethylcytosine | 0.23 | 0.00 |
| Methyl isoamylene methyl billetier | 0.40 | 0.00 |
| New silane neoussilagine | 0.18 | 0.00 |
| DG(20:1(11Z)/15:0/0:0) | 2.04 | 0.00 |
| 1-[(2-chlorophenyl)methylsulfonyl]N-cyclophosphatidine-3-carboxamide for improving antioxidant substances | 2.54 | 0.01 |
| 13-hydroxy abscisic acid 13-Hydroxyabscisic acid | 0.35 | 0.00 |
| 3'-hydroxy-3,4,5,4' -tetramethyleneOxystilbene 3'-Hydroxy-3,4,5,4' -tetramethoxyysilbene | 0.41 | 0.00 |
| Serine lysine seryleysine | 3.09 | 0.01 |
| 3-Deoxy-D-glycero-D-galacto-2-non-fructo-acid 3-Deoxy-D-glyco-D-galactose-2-nonnulosonic acid | 0.38 | 0.00 |
| Blue enol C glucoside Blumenol C glucoside | 2.11 | 0.03 |
Note that: FC (E/C) is the fold difference in expression (fold change) of the metabolite between the two groups.
C: the expression level of the metabolite in the control group,
e: the expression level of the metabolite in the experimental group,
TABLE 7 short chain fatty acids
| Acetic acid | Propionic acid | Isobutyric acid | Butyric acid | Isopentanoic acid | Valeric acid | Isohexanoic acid | Caproic acid | |
| Test group | 5.13±0.74 | 0.09 A ±0.01 | 0.02 A ±0.00 | 0.03 A ±0.02 | 0.01 A ±0.00 | 0.01 A ±0.00 | 0.21 b ±0.05 | 0.01 A ±0.00 |
| Control group | 4.86±0.82 | 3.19 B ±0.62 | 0.06 B ±0.02 | 1.56 B ±0.31 | 0.05 B ±0.01 | 0.44 B ±0.27 | 0.16 a ±0.01 | 0.17 B ±0.10 |
Propionic acid is generally lower in the green feed, and the content of propionic acid, isobutyric acid, isovaleric acid, valeric acid, caproic acid and butyric acid is extremely obviously reduced and the content of isocaproic acid is greatly increased after the patent bacteria are added in the study. Butyric acid is closely related to clostridium butyricum, which aggravates the loss of dry matter and causes the rise of acetic acid, soluble protein and ammonium nitrogen, the content of which is inversely proportional to the quality of silage; the isohexide content is proportional to the digestibility of ruminants such as cows, and in the above examples, the content of butyric acid is reduced and the content of isohexide is increased after the addition of the patent bacteria, which indicates that the quality of silage is improved.
The foregoing has shown and described the basic principles, principal features and advantages of the invention. It will be understood by those skilled in the art that the foregoing test examples and descriptions are merely illustrative of the principles of the present invention and that various changes and modifications may be made therein without departing from the spirit and scope of the invention, the scope of which is defined in the appended claims, specification and their equivalents.
Claims (9)
1. Special acid-feeding dyford bacterial strain for ensiling of bromhidrosisDelftia acidovorans) The method is characterized in that: the preservation name of the special delfurd bacterium strain for ensiling the bromhidrosis is delfurd bacterium LL2022006; the preservation unit is China general microbiological culture Collection center (CGMCC), and the preservation address is North Xishlu No.1 and No. 3 in the Korean area of Beijing; preservation date: 2022, 4, 7; the preservation number is CGMCC NO.24652.
2. The special acid-feeding dyford bacterial strain for ensiling of foxtail algae according to claim 1, characterized in that: the 16S rDNA gene sequence of the special acid-feeding dyford bacterial strain for ensiling the bromhidrosis is a nucleotide sequence shown as SEQ ID No. 1.
3. The special acid-feeding dyford bacterial strain for ensiling of foxtail algae according to claim 1, characterized in that: the colony morphology and gram staining microscopic examination are characterized in that: in MRS agar culture medium, after aerobic culture for 72 hours at 35 ℃, bacterial colony is milky round, smooth in surface, neat in edge and easy to pick up; gram staining is positive, and the thalli are rod-shaped, single or double, and are facultative anaerobic.
4. The microbial inoculum prepared by the special acid-feeding dyford bacteria for ensiling the foxtail algae according to claim 1.
5. The microbial inoculum of claim 4, wherein the active ingredient is at least one of the following (a) (b) (c):
(a) The fermentation culture of the special acid-feeding dyford microbial inoculum for ensiling the foxtail algae of claim 1;
(b) The spore suspension of the special dyford bacterium agent for ensiling the bromhidrosis obtained in the claim 1;
(c) Ultrasonic lysis and precipitation of special acid-feeding dyford microbial inoculum cells for ensiling the bromhidrosis obtained in claim 1.
6. The method for preparing the microbial inoculum of claim 4, which is characterized by comprising the following steps: (1) strain preparation: inoculating v=0.1% in MRS broth culture medium, culturing at 35-37 ℃ for 36~48 h,5000~8000 rpm, and centrifuging to obtain thalli;
(2) Freeze-drying the thalli to prepare the special acid-feeding dyford fungus preparation for ensiling the foxtail algae.
7. The method for preparing the microbial inoculum according to claim 6, characterized in that: in step (1), the MRS medium broth is: 10.0g of peptone; beef extract 5.0g; 4.0g of yeast powder; glucose 20.0g; 2.0g of dipotassium hydrogen phosphate; 2.0g of tri-ammonium citrate; 5.0g of sodium acetate; 0.2g of magnesium sulfate; manganese sulfate 0.05g; tween 80.0 ml; 1L of water.
8. The method for preparing the microbial inoculum according to claim 6, characterized in that: in the step (2), the effective viable count of the bacterial cells in the microbial preparation is 10 10 ~10 11 CFU/g。
9. The use of the acid-feeding dyford bacterium of claim 2 or the microbial inoculum of claim 4 in the preparation of feed.
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| CN104531585A (en) * | 2014-12-31 | 2015-04-22 | 中国水稻研究所 | Delftia tsuruhatensis and application thereof |
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| CN1782069A (en) * | 2005-10-26 | 2006-06-07 | 广东绿百多生物科技有限公司 | Method for cultivating acide eating Darfot bacteria using combustible gas sensor fermentation |
| CN104531585A (en) * | 2014-12-31 | 2015-04-22 | 中国水稻研究所 | Delftia tsuruhatensis and application thereof |
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