CN116410897A - Friedson escherichia and antibacterial application thereof - Google Patents
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- CN116410897A CN116410897A CN202310377947.3A CN202310377947A CN116410897A CN 116410897 A CN116410897 A CN 116410897A CN 202310377947 A CN202310377947 A CN 202310377947A CN 116410897 A CN116410897 A CN 116410897A
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Abstract
The invention belongs to the technical field of biology, and particularly relates to a Frugsen Escherichia bacterium and an antibacterial application thereof. According to the invention, 1 strain of the Fr-Glen escherichia 306 is separated and identified from animal probiotics products, and then antibacterial activity research is carried out on the Fr-Glen escherichia by taking a produced biosurfactant product as an entry point. The strain of the Freeze escherichia 306 is found to generate a biosurfactant product, and the strain and the metabolite thereof have certain antibacterial effect on staphylococcus aureus and escherichia coli. Meanwhile, the 306 strain is sensitive to common antibacterial drugs in animal clinic, and has no pathogenicity and toxic and side effects. Therefore, the Fuglesen strain 306 and the metabolite thereof developed by the invention have potential application value in developing into animal antibiotic additive products.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a Frugsen Escherichia bacterium and an antibacterial application thereof.
Background
The Fr-Glen Escherichia bacterium (Escherichia fergusonii) belongs to the genus Escherichia of the family Enterobacteriaceae. The Fleens are generally considered to be conditional pathogenic bacteria, have close relationship with diseases of animals and humans, and are pathogenic and virulent to both humans and animals. In recent years, the fgen escherichia bacteria are resistant to antibacterial drugs for treating intestinal bacterial infections, and the resistant phenotype is similar to escherichia coli. There are studies to isolate a strain of fgison in a cystitis patient and to find that the pathogenic strain has resistance to various antibiotics by measuring the Minimum Inhibitory Concentration (MIC) of the different antibiotics against the strain. In one cystitis patient case report in 2010, a strain of fgrass, which expresses ultra-broad spectrum beta-lactamase (ESBL) and is resistant to various antibiotics, was isolated clinically for the first time, and the strain exhibited high resistance to common drugs of cephalosporins, quinolones, and sulfonamides.
In recent years, research into the strain of the genus fgrass has been focused mainly on the carrying of drug resistance genes, and some strains of the strain fgrass are considered as "reservoirs" of drug resistance genes. 2021, russian federal scholars reported that the detection of genetic association of toxicity of the friesen escherichia and resistance to antibacterial agents allowed the inclusion of friesen escherichia pathogens that pose a threat to public health. In the same year, brazilian scholars separated 3 strains of Frugreek from the cloacal cotton swab of broilers, and the genome of the 3 strains of Frugreek comprises four plasmids, wherein one plasmid is an IncHI2 plasmid of 182869bp, and the plasmid contains a drug resistance gene mcr-1. Meanwhile, it was found that the total genome length of the strain of the fgrass strain OTSVEF-60 from the national fowl sample of mangladesh was 4.2Mbp, comprising 4503 coding sequences, and 41 antibiotic resistance genes (ACG) were distributed. It has also been found that the whole genome of colistin-resistant Escherichia coli EFCF056 isolated from chicken manure contains 6 plasmids, one of which is a plasmid containing colistin-resistant gene mcr-1. In addition, there are studies in which 22 metal resistance genes are found in the genome of the POEF strain.
In 2021, indian students separated and identified the strain of Freyn Escherichia AMC01 from intestinal tracts of black abdomen tomb bat by using a chitin agar medium, and the strain has chitin dissolving activity and is expected to be an effective strain for producing chitinase. Meanwhile, there have been studies on the identification of chitinase-producing Escherichia friendliensis from calf feces having chitin-dissolving activity. In addition, there is a study on screening 1 strain of a Furthern Escherichia coli LW-2 strain producing succinic acid at a high yield by using bovine rumen content as a bacterial source and sodium fumarate as a carbon source. There are studies on separating and identifying a strain of the Escherichia fraGlen which can effectively repair textile wastewater from the wastewater.
In summary, the friendlien escherichia bacteria from different separation sources have different biological characteristics, and the excavation of more friendlien escherichia bacteria with different biological activities has important application prospects. However, there has been no report of the antibacterial activity of the fgrass bacteria.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention provides the Fr-Glen escherichia with certain antibacterial effect on staphylococcus aureus and escherichia coli, which has potential application value in developing an animal antibacterial additive product, and widens the application range of the Fr-Glen escherichia.
In order to achieve the above purpose, the present invention is realized by the following technical scheme:
in a first aspect, the invention provides a method for producing a strain of the genus Fungessen, wherein the strain is Escherichia fergusonii, deposited at the Cantonese microorganism strain collection at about 21/02/2023 under the accession number GDMCC No. 63180.
The second aspect of the invention provides an application of the Frugreek bacterium in preparing a bacteriostatic agent.
In a third aspect, the invention provides the use of the friendliensin bacterium of the first aspect in the preparation of a biosurfactant.
The strain of the genus fgison (Escherichia fergusonii) is generally considered a conditional pathogen and plays an important role in the transmission of the resistance of enteropathogenic bacteria, and some strains of the strain of the genus fgison are "reservoirs" of the resistance genes. Meanwhile, the friendliensis strain isolated from a specific environment has the characteristics of specific substrate degradation, metal salt and dye resistance and stress-resistant growth, and is attracting attention in environmental bioremediation. In the invention, 1 strain of the Fuglesen strain 306 is separated and identified from animal probiotics products. And performing antibacterial activity test research on the Fr-Glen escherichia with the biosurfactant as an entry point. The results show that: the strain of the Freeze escherichia 306 can generate a biosurfactant product, and the strain and the metabolite thereof have certain antibacterial effects on staphylococcus aureus and escherichia coli, wherein the strain 306 has strong antibacterial effect on the staphylococcus aureus. Meanwhile, the drug sensitivity test results show that: the 306 strain is sensitive to common antibacterial drugs in animal clinic. In addition, the bacterial liquid is fed to the mice without pathogenicity and toxic side effects. The test results suggest that: the strain of the Freeze escherichia coli 306 is a potential animal probiotics, and the metabolite of the strain has important application value in the aspect of developing new animal antibacterial peptide additive products.
Preferably, the inhibition is of staphylococcus aureus and/or of escherichia coli.
Preferably, the biosurfactant is an anionic biosurfactant.
In a fourth aspect, the present invention provides a bacteriostatic agent comprising the escherichia freguessen as the main active ingredient.
According to a fifth aspect of the present invention, there is provided a method for preparing the bacteriostatic agent according to the fourth aspect, wherein the method comprises the steps of performing liquid fermentation by using the escherichia fraGlon according to the first aspect, and collecting the product to obtain the bacteriostatic agent.
Preferably, the temperature of the liquid fermentation is 37 ℃, the time is 24 hours, the rotating speed is 120 revolutions per minute, and the culture medium of the liquid fermentation is LB culture medium.
Preferably, the collecting the product further comprises the steps of: 1mL of the obtained product was centrifuged at 10000rmp for 10min, and the supernatant was collected.
More preferably, the collecting the product further comprises the steps of: 1mL of the supernatant was filtered through a 0.45. Mu.mL filter, and the filtrate was collected.
Compared with the prior art, the invention has the beneficial effects that:
according to the invention, 1 strain of the Fr-Glen escherichia 306 is separated and identified from animal probiotics products, and then antibacterial activity research is carried out on the Fr-Glen escherichia by taking a produced biosurfactant product as an entry point. The strain of the Freeze escherichia 306 is found to generate a biosurfactant product, and the strain and the metabolite thereof have certain antibacterial effect on staphylococcus aureus and escherichia coli. Meanwhile, the 306 strain is sensitive to common antibacterial drugs in animal clinic, and has no pathogenicity and toxic and side effects. Therefore, the Fuglesen strain 306 and the metabolite thereof developed by the invention have potential application value in developing into animal antibacterial additive products, and widen the application range of the Fuglesen strain.
Drawings
FIG. 1 shows colony morphology of strain Fragrantis 306;
FIG. 2 shows the result of gram staining of the Freeze Escherichia bacterium;
FIG. 3 is a genetic phylogenetic tree of the strain Fragrantis Escherichia coli 306;
FIG. 4 shows the results of an emulsion test of the French Escherichia bacterium 306;
FIG. 5 shows the experimental results of the C-TAB agar test (filter paper method) for strain Freeze 306 (1-6 is a faint blue halo around the lawn of strain 306, 7 is a 20% Tween 80 positive control);
FIG. 6 is a comparative experiment (1-6 is beta hemolysis around the lawn of strain 306, 7 is 20% Tween 80 positive control) of the French Escherichia strain 306 blood agar plate hemolysis test (filter paper method);
FIG. 7 shows the results of an antibacterial test of strain Fragrantis 306, with Staphylococcus aureus on the left and Escherichia coli on the right; 1.2 and 3 are respectively the bacterial liquid, the supernatant and the filtrate of the strain of the Freeze escherichia coli 306, and 4, 5 and 6 are respectively 1, 2 and 3.7 (left) is Doxycycline (DO) control and 7 (right) is neomycin (N) control.
Detailed Description
The following describes the invention in more detail. The description of these embodiments is provided to assist understanding of the present invention, but is not intended to limit the present invention. In addition, the technical features of the embodiments of the present invention described below may be combined with each other as long as they do not collide with each other.
The experimental methods in the following examples, unless otherwise specified, are conventional, and the experimental materials used in the following examples, unless otherwise specified, are commercially available.
EXAMPLE 1 isolation and identification of Fuglesen Escherichia bacteria and antibacterial Activity test study
1. Materials and methods
1.1 test materials
1.1.1, culture medium: LB medium (g/L): glucose 10, yeast powder 5, naCl 10, peptone 10 and agar 17 (added in solid culture medium), adjusting pH to 7.0, sterilizing at 121deg.C for 20min for use. Cetyl trimethylammonium bromide (C-TAB) medium: to the liquid LB medium, C-TAB (0.5 mg/mL) and methylene blue (0.2 mg/mL) were added and the corresponding agar was added. MRS Medium (g/L): peptone 10, beef extract powder 5, yeast extract powder 4, glucose 20, K 2 HPO 4 2, sodium acetate 5, triammonium citrate 2, mgSO 4 .7H 2 O 0.2，MnSO 4 .4H 2 O0.05, agar 17 (solid culture medium is added), finally adding 1mL Tween 80, adjusting pH to about 6.2, sterilizing at 121deg.C for 20min for use. Blood agar medium: purchased from Guangdong Crypton microbiological technologies Co. Bacterial biochemical reaction reagent: purchased from Guangdong Crypton microbiological technologies Co. Drug sensitive test paper sheet: purchased from Guangdong Crypton microbiological technologies Co.
1.1.2, indicator bacteria: coli (K12D 31), staphylococcus aureus (ATCC 29213) were from the chinese collection of microorganisms.
1.1.3, mice for test and feed: mice NIH, SPF grade, were purchased from the medical laboratory animal center in guangdong province.
1.2, test methods
1.2.1 isolation of the Fuglesen Escherichia bacteria
The purpose of the original test is to separate bacillus strains with bacteriostasis from probiotic fermentation products, and the specific steps are as follows:
(1) 5 g of the product [ BAIJUAN MENTONIAN lyophilized powder (Q/hxswj 001-2018) of probiotic additive purchased from Bofeng Biotechnology Co., guangzhou ] was taken, 5mL of sterile physiological saline was added, fermentation was performed in a test tube at 37℃for 6 hours, 4500prm was taken out, and the supernatant was taken after centrifugation, and heated at 60℃for 30 minutes, to obtain a Bacillus separation solution.
(2) Progressively diluting bacillus separating liquid by 10 times, and taking 10 times 7 And 10 8 The tube is used as a sample for separating bacteriostatic bacillus, and a LB plate of staphylococcus aureus indicator bacteria is used for separating bacteriostatic strains. Instant 10 7 And 10 8 The tube samples were applied to LB plates of staphylococcus aureus indicator bacteria in 0.2mL, cultured at 37℃for 24 hours, and bacterial colonies with a bacteria inhibition ring around the bacterial colonies were observed to isolate bacteria-inhibiting bacillus.
(3) The observation of the colony found 1 unknown strain with bacteriostasis ring around the colony with diameter of 1.5-2mm and smooth surface and moist circular colony. This colony was then further purified and designated as 306 strain, with subsequent identification and bacteriostasis analysis.
1.2.2, strain culture and filtrate preparation: staphylococcus aureus and Escherichia coli were inoculated into LB medium, respectively, and cultured with shaking (120 rpm) at 37℃for 16-24 hours, at a bacterial concentration (OD 600nm ) About 0.1, and is taken out and placed at 4 ℃ for standby.
1.2.3 culture of Fuglesen Escherichia bacteria: the strain of the prepared Fr-Glen Escherichia bacteria is separated and cultured by using a common agar plate, a typical colony is selected, inoculated in an LB culture medium, cultured for 24 hours by shaking at 37 ℃ (120 revolutions per minute), and a bacterial solution is taken out and placed at 4 ℃ for standby. Preparing supernatant: 1mL of bacterial liquid is taken and added into a centrifuge tube, 10000rmp is centrifuged for 10min, and supernatant is taken and placed at 4 ℃ for standby. And (3) preparing a filtrate: 1mL of the supernatant was collected, filtered through a 0.45. Mu.mL filter, and the filtrate was collected and left at 4℃for further use.
1.2.4 method for detecting biological surface Activity
(1) Emulsifying activity (Emulsifying activity, EA): after 2.5mL of the 24-hour culture of the test bacteria was taken aseptically and centrifuged at 10,000r/pm, 2mL was pipetted into a 5mL centrifuge tube, and an equal amount of petroleum ether was added to the sample as the test oil phase, and the mixture was vigorously vortexed for 2min. After stabilization for 1min, the emulsion layer height was divided by the total height of the mixture, multiplied by 100, and the emulsifying capacity was measured and calculated.
(2) Emulsion index detection (E24 index detection, E24): after the emulsion activity detection tube was left to stand at 4℃for 24 hours, the measured height of the emulsion layer was divided by the total height of the mixture, and multiplied by 100 to calculate the emulsion index (E24).
(3) Hemolytic Activity (Hemolytic activity): the test bacteria were inoculated by a conventional method of inoculating the bacteria on an agar plate and a method of inoculating a filter paper sheet (8 uL of 20% Tween 80 as a positive control), and after inoculation, incubated at 37℃for 24 hours. The colonies or the hemolyzed halos around the inoculated filter paper sheets were removed, and the production intensity of the Biosurfactant (BS) was determined.
(4) TAB detection: cetyl trimethyl ammonium bromide (C-TAB) agar plate method is a rapid screening detection method for detecting anionic Biosurfactants (BSs). Bacterial solutions (also seeded with 8uL of 20% tween 80 as positive control) were inoculated in a haemolytic activity inoculation (filter paper) using cetyltrimethylammonium bromide (C-TAB) agar plates, each at about 10uL, incubated for 24 hours at 37 ℃ and then stored at 4 ℃ for further development of colour (24 hours). The presence of a dark blue halo around the surface flora of the C-TAB agar plate was judged as a positive result, and the absence of a dark blue halo was judged as a negative result.
1.3, entrusting Guangzhou Ai Ji biotechnology limited company to carry out 16S rDNA sequencing, and comparing the sequence homology of the 16S rDNA gene fragment sequencing result with a gene BANK database to establish a strain 16S rDNA genetic development tree.
1.4 bacteriostasis test
1.4.1, preparation of bacteria inhibition flat plate of indicator bacteria: taking LB solid culture medium cooled to 50-56 ℃ after dissolution, adding about 20 mu L of indicator bacteria liquid into each plate, fully and uniformly mixing, pouring the plates, and carrying out sterile air drying on surface condensed water for later use.
1.4.2, bacteriostasis method: (1) determining the antibacterial activity of the bacterial liquid by adopting a filter paper sheet method: taking out the bacterial liquid to be detected, attaching a filter paper sheet (with the diameter of 6mm, sterilizing at 121 ℃ for 20min and storing in a drying oven at 60 ℃ for later use) to a preset position, taking out the bacterial liquid to be detected (about 10 mu L) to be dripped on the filter paper sheet attached to the surface of an indicator bacteria plate, taking a neomycin drug sensitive sheet (an escherichia coli indicator bacteria plate) and a culture medium control (a staphylococcus aureus indicator bacteria plate), culturing at 37 ℃ for 24 hours, taking out, measuring the diameter (mm) of a bacteriostasis ring by using a vernier caliper, and recording the result. (2) The antibacterial activity of the bacterial liquid extract, the supernatant and the filtrate is measured by adopting an agar diffusion method: punching with a puncher with the diameter of 6mm according to a preset position, picking out agar in the hole, taking the tested bacterial liquid (about 80 mu L), dripping the tested bacterial liquid into the hole until the bacterial liquid is fully filled and does not overflow, and otherwise, the method is the same as a filter paper sheet method. And (3) placing the sample in an incubator at 37 ℃ for 24 hours, taking out the sample, observing the result, measuring the diameter (mm) of the inhibition zone by using a vernier caliper, and recording the result.
1.5 safety feeding test for mice
Mice purchased from the medical laboratory center were fed to the animal room for 2 days and then divided into test and control groups of 7 animals each. On the first day of the test group, each mouse was filled with 0.1mL of bacterial liquid, and then the bacterial liquid was added to the test group drinking water at an amount of 0.1 mL/day per mouse, and the test group was taken freely until the 7 th day (normal diet of the control group). The mental state, the eating state, the normal or abnormal feces, the morbidity or mortality and the like of the mice are observed every day. Test and control mice were weighed 2 times on day 3 and day 7, respectively, and the results were recorded.
2. Experimental results
2.1 colony morphology, staining, biochemical identification results
After the strain of the French Escherichia 306 is subjected to streak culture (37 ℃ C., 24 h) by a common agar plate, the colony is off-white, round or nearly round, the diameter is 3-4 mm, the surface is drier and flat, the middle of the colony is slightly raised, and the periphery is semitransparent (figure 1). Meanwhile, the 306 strain is cultured for 24 hours at 37 ℃ by a common agar plate, smeared, gram stained and microscopic examination. As a result, it was found (FIG. 2) that the gram stain of the Freeze-on Escherichia coli 306 strain was negative, which was a relatively short, thick, medium-sized bacterium. In addition, the bacterial biochemical reactions of the strain fgrass 306 are shown in table 1. Table 1 further demonstrates that strain 306 belongs to the genus Fragrantis from the standpoint of bacterial biochemical reactions.
TABLE 1 Biochemical reaction identification of Ehrlichia feculence 306 Strain
| Authentication item | 306 strain | Authentication item | 306 strain |
| Glucose | + | 6.5% high salt broth | Growth |
| Lactose and lactose | － | Malonic acid salt | － |
| Fructose | + | Citrate salt | － |
| Maltose | － | Nitrate reduction | － |
| Mannitol (mannitol) | － | Starch hydrolysis | － |
| Sucrose | + | Phenylalanine (Phe) | － |
| Cellobiose | + | Arginine hydrolysis | + |
| Mushroom candy | + | H 2 S generation | － |
| Honey diol | － | Motility of | + |
| Rhamnose (rhamnose) | － | N-acetylgluglucosaminium | － |
Remarks: a "+" positive response; "-" negative reaction.
2.2 identification of 16S rDNA Gene sequence of Fuglessen Escherichia coli 306 Strain and genetic development Tree
By sequencing the 16S rDNA gene fragment and comparing the sequence homology with a gene BANK database, the 306 strain is found to be the Fuglesen Escherichia bacterium, the genetic phylogenetic tree of the 306 strain is shown as figure 3, and the 306 strain is confirmed to be a new strain in the Fuglesen Escherichia bacterium.
Finally, the strain of the Escherichia coli 306 is preserved, and the preservation information is as follows:
preservation time: 2023, 02, 21;
preservation unit name: the collection of microorganism strains in Guangdong province;
preservation number: GDMCC No. 63180;
deposit unit address: guangzhou city first middle road No. 100 college No. 59 building 5;
classification naming: escherichia fergusonii 306, 306.
The 16S rDNA sequence of Escherichia fergusonii (E.fergusoni 306) is as follows:
CGCTGACGAGTGGCGGACGGGTGAGTAATGTCTGGGAAACTGCCTGATGGAGGGGGATAACTACTGGAAACGGTAGCTAATACCGCATAACGTCGCAAGACCAAAGAGGGGGACCTTCGGGCCTCTTGCCATCGGATGTGCCCAGATGGGATTAGCTAGTAGGTGGGGTAACGGCTCACCTAGGCGACGATCCCTAGCTGGTCTGAGAGGATGACCAGCCACACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGCAAGCCTGATGCAGCCATGCCGCGTGTATGAAGAAGGCCTTCGGGTTGTAAAGTACTTTCAGCGGGGAGGAAGGGAGTAAAGTTAATACCTTTGCTCATTGACGTTACCCGCAGAAGAAGCACCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGCACGCAGGCGGTTTGTTAAGTCAGATGTGAAATCCCCGGGCTCAACCTGGGAACTGCATCTGATACTGGCAAGCTTGAGTCTCGTAGAGGGGGGTAGAATTCCAGGTGTAGCGGTGAAATGCGTAGAGATCTGGAGGAATACCGGTGGCGAAGGCGGCCCCCTGGACGAAGACTGACGCTCAGGTGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGTCGACTTGGAGGTTGTGCCCTTGAGGCGTGGCTTCCGGAGCTAACGCGTTAAGTCGACCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAATGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGATGCAACGCGAAGAACCTTACCTGGTCTTGACATCCACGGAAGTTTTCAGAGATGAGAATGTGCCTTCGGGAACCGTGAGACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTTGTGAAATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATCCTTTGTTGCCAGCGGTCCGGCCGGGAACTCAAAGGAGACTGCCAGTGATAAACTGGAGGAAGGTGGGGATGACGTCAAGTCATCATGGCCCTTACGACCAGGGCTACACACGTGCTACAATGGCGCATACAAAGAGAAGCGACCTCGCGAGAGCAAGCGGACCTCATAAAGTGCGTCGTAGTCCGGATTGGAGTCTGCAACTCGACTCCATGAAGTCGGAATCGCTAGTAATCGTGGATCAGAATGCCACGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTGCAAAAGAAGTAGGTAGCTTAACCTTCGGGAGGGCGCT。
2.3 drug sensitivity test and mouse feeding safety test of the Freeze Escherichia coli 306 Strain
The results of the drug susceptibility test of the strain Freeze-on Escherichia coli 306 are shown in Table 2. As can be seen from Table 2, the strain of Fragrantis 306 is sensitive to common antibacterial agents in animal clinics, indicating that the strain of Fragrantis 306 is not resistant and transmits resistance.
Mice were fed with the fgen escherichia 306, and during the test period, mice were well-conditioned, normally on diet, normally on faeces, without morbidity and mortality, and without differences in weight gain between the test group and the mice compared to the control group. Indicating that the Freeze Escherichia coli 306 has no pathogenicity and toxic and side effects.
Table 2 results of drug sensitivity test of the fgisenia strain 306 the diameter of the zone of inhibition: mm (mm)
| Test strains/drugs | Amoxicillin | Amikacin | Gentamicin | Ciprofloxacin | Star enrofloxacin |
| Friedson escherichia 306 | 22 | 19 | 18 | 30 | 32 |
| Coli K12D31 | 10 | 20 | 18 | 28 | 28 |
| Test strains/drugs | Florfenicol | Neomycin | Doxycycline | Ceftriaxone | Compound Xinnoming |
| Friedson escherichia 306 | 24 | 14 | 30 | 26 | 23 |
| Coli K12D31 | 27 | 13 | 26 | 26 | 26 |
2.4 biological surface Activity test results of Fuglessen Escherichia coli 306 Strain
As shown in fig. 4, 5, and 6, the emulsion index (EA) of the fgese escherichia coli 306 was 50%, and after 24 hours (E24), it was 28%. After incubation with C-TAB medium, blue circular halos are formed around the filter paper sheet and lawn. The 306 strain is separated and cultured by blood agar plate streaking (37 ℃ for 48 hours), and the periphery of the lawn is subject to beta hemolysis.
2.5 antibacterial test against Freyn Escherichia coli
Qualitative antibacterial tests were performed on the strain fgison escherichia 306, and the results show (fig. 7): the Fugleen escherichia 306 strain, the supernatant and the filtrate have inhibition effects on staphylococcus aureus and escherichia coli, wherein the Fugleen escherichia 306 strain has strong bacteriostasis on the staphylococcus aureus and weak bacteriostasis on the escherichia coli.
In conclusion, the Fuglen escherichia 306 strain has a certain inhibition effect on indicator bacteria of gram-positive staphylococcus aureus and gram-negative escherichia coli, is a potential animal probiotic, and the metabolite of the Fuglen escherichia 306 strain has important application value in the aspect of developing new animal antibacterial peptide additive products.
The embodiments of the present invention have been described in detail above, but the present invention is not limited to the described embodiments. It will be apparent to those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, and yet fall within the scope of the invention.
Claims (10)
1. The Frugreek strain is Escherichia fergusonii and is deposited at the microorganism strain collection of Guangdong province at the month 21 of 2023, and the deposition number is GDMCC No. 63180.
2. The use of the fgen escherichia of claim 1 in the preparation of a bacteriostatic agent.
3. Use of the friendliensis of claim 1 for the preparation of a biosurfactant.
4. Use according to claim 2, wherein the inhibition is of staphylococcus aureus and/or of escherichia coli.
5. Use according to claim 2, wherein the biosurfactant is an anionic biosurfactant.
6. A bacteriostatic agent characterized by comprising the escherichia frangensis of claim 1 as a main active ingredient.
7. The method for preparing the antibacterial agent according to claim 6, characterized in that the antibacterial agent is obtained by liquid fermentation of the friendliensiella according to claim 1 and collecting the product.
8. The method for preparing a bacteriostat according to claim 7, wherein the liquid fermentation temperature is 37 ℃, the time is 24 hours, the rotating speed is 120 revolutions per minute, and the liquid fermentation culture medium is LB culture medium.
9. The method for preparing a bacteriostatic agent according to claim 7, wherein said collecting the product further comprises the steps of: 1mL of the obtained product was centrifuged at 10000rmp for 10min, and the supernatant was collected.
10. The method of preparing a bacteriostatic agent according to claim 9, wherein said collecting the product further comprises the steps of: 1mL of the supernatant was filtered through a 0.45. Mu.mL filter, and the filtrate was collected.
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2023
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN118161534A (en) * | 2024-05-14 | 2024-06-11 | 上海交通大学医学院附属上海儿童医学中心 | Fuglesen escherichia and use of product thereof in inflammatory diseases |
| CN118161534B (en) * | 2024-05-14 | 2024-08-20 | 上海交通大学医学院附属上海儿童医学中心 | Fuglesen escherichia and use of product thereof in inflammatory diseases |
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