CN116355788A - Wittman coagulans GBW0011 and application thereof in preparation of oral bacteriostat - Google Patents
Wittman coagulans GBW0011 and application thereof in preparation of oral bacteriostat Download PDFInfo
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Abstract
The invention discloses a strain of Willemann condensation GBW0011 and application thereof in preparation of an oral bacteriostat. The classification of the condensation Wittman's GBW0011 is named as condensation Wittman's Weizmannia coagulans, and the preservation number is CCTCC NO: m20221404 the strain has the capability of inhibiting the reproduction of oral pathogenic bacteria, namely streptococcus mutans and porphyromonas gingivalis, and the combination of the Willebrand coagulation GBW0011 and chlorhexidine can effectively reduce the expression level of gingival crevicular fluid inflammatory factors IL-1 beta and TNF-alpha mRNA of a patient suffering from gingival bleeding, improve the venous blood coagulation function of the patient suffering from gingival bleeding, and further improve the treatment effect on the patient suffering from gingival bleeding.
Description
Technical Field
The invention belongs to the field of microorganisms, and particularly relates to a strain of condensation Wittman's GBW0011 and application thereof in preparation of oral bacteriostat.
Background
Most of the reasons for gingival bleeding are caused by local factors, such as dental plaque, calculus, bad restoration or food residues on the neck of teeth, which stimulate gingival edema congestion and cause local capillary rupture, and the bleeding part is mostly positioned at the papilla and gingival sulcus, so that the purpose of rapid hemostasis is difficult to achieve by a general hemostasis method, and the gingival bleeding is often accompanied with halitosis, obstructs work social activities and brings heavy mental burden to patients.
Currently, the focus of the clinical treatment of gingival bleeding is mainly to eliminate pathogenic bacteria, and bacterial plaque inhibition is carried out through some effective medicines, such as borax gargle, chlorhexidine gargle and the like. Borax gargle has pungent smell, and has poor curative effect due to lower compliance of patient treatment. The chlorhexidine collutory has good plaque inhibition effect, is fragrant in smell and more acceptable to patients, and is a treatment mode which is more commonly used for gingival bleeding in recent years. However, previous studies have shown that patients still have a high risk of recurrence after chlorhexidine treatment, which is closely related to the colonization of pathogenic bacteria in the patients oral cavity.
As the oral cavity has more microorganisms, and pathogenic bacteria such as streptococcus mutans, porphyromonas gingivalis and the like grow rapidly. Research shows that the administration of a certain exogenous probiotics in the treatment of gingival bleeding patients can inhibit the reproduction of oral pathogenic bacteria, is beneficial to maintaining the ecological balance of the oral cavity and effectively consolidates the treatment effect.
Disclosure of Invention
The invention aims to provide a strain of condensation Wittman GBW0011 and application thereof in preparation of oral bacteriostat. The M.Weizhenmann condensation GBW0011 has the capability of inhibiting the reproduction of streptococcus mutans and porphyromonas gingivalis.
In order to achieve the above purpose, the invention adopts the following technical scheme:
the invention provides a strain of Wettman coagulans GBW0011, which is classified and named as Wettman coagulans Weizmannia coagulans, and has a preservation number of CCTCC NO: m20221404.
Further, the nucleotide sequence of the 16S rRNA of the GBW0011 of the M.coagulans is shown as SEQ ID No. 1; the nucleotide sequence of adk is shown in SEQ ID No. 2.
The invention also provides application of the condensation Wittman GBW0011 in preparation of an oral bacteriostatic agent.
Furthermore, the oral bacteriostat contains the cell-free filtrate of the condensation Wittman's GBW0011 with the concentration of 0.5% -2.0% (v/v).
Further: the concentration of the clear filtrate of the Wittman coagulans GBW0011 is 1% (v/v), and the clear filtrate has a killing effect on streptococcus mutans; the concentration of the Wittman coagulans GBW0011 cell-free filtrate is 1.5% (v/v), and the water-based emulsion has a killing effect on Porphyromonas gingivalis.
Further, the preparation method of the cell-free filtrate of the GbW0011 of the Willebrand-new strain comprises the following steps: the activated Welch mannomyces coagulans is inoculated into a fermentation culture medium at an inoculation ratio of 2% -5% (v/v), and is cultured for 24-36 h at 37 ℃ under anaerobic condition, and the obtained culture solution is centrifuged and filtered to obtain the supernatant, namely the Welch mannomyces coagulans GBW0011 cell-free filtrate.
Further, the formula of the fermentation medium is as follows: 5 to 8 percent of soybean meal, 1 to 2 percent of peptone, 1 to 3 percent of corn flour, 1 to 1.5 percent of glucose, 0.3 to 0.5 percent of dipotassium hydrogen phosphate, 0.02 to 0.1 percent of manganese sulfate and 0.05 to 0.1 percent of sodium nitrate.
Further, the pathogenic bacteria inhibited by the oral bacteriostat are streptococcus mutans and porphyromonas gingivalis.
The invention also provides application of the condensation Wittman GBW0011 in preparation of a medicament for preventing and treating gingival bleeding.
Furthermore, the condensation Wittman GBW0011 can effectively reduce the expression level of inflammatory factors in gingival crevicular fluid and improve the blood coagulation function, thereby achieving the purpose of preventing and treating gingival bleeding.
The invention also provides a liquid containing a rinse, the effective component of the liquid containing the liquid is bacteria content of not less than 1.0X10 6 CFU/mL of GBW0011 bacterial solution of M.Weizhengensis and chlorhexidine.
Compared with the prior art, the invention has the advantages and technical effects that:
the invention provides a Wettman-coagulating GBW0011, which has the capability of inhibiting reproduction of oral pathogenic bacteria streptococcus mutans and Porphyromonas gingivalis, can reduce the expression level of gingival crevicular fluid inflammatory factors IL-1 beta and TNF-alpha mRNA of a gingival bleeding patient when the Wettman-coagulating GBW0011 is combined with chlorhexidine, improves venous blood coagulation function of the gingival bleeding patient, improves treatment effect on the gingival bleeding patient, and provides theoretical basis for practical application of the Wettman-coagulating.
Drawings
FIG. 1 shows colony morphology of M.Weizhengensis GBW0011 on MRS plates.
FIG. 2 shows the morphological characteristics of spores of GbW0011 of M.weiarrowhead under a microscope.
FIG. 3 shows the growth of Streptococcus mutans and Porphyromonas gingivalis in different concentrations of Wittman's coagulation cell-free filtrate.
Detailed Description
The technical scheme of the invention is further described in detail below with reference to specific embodiments. In the following examples, unless otherwise specified, all experimental methods used are conventional and all materials, reagents, etc. are commercially available from biological or chemical reagent companies.
Example 1: screening, separation and identification of Wittman coagulans GBW0011
1. Screening and purification of Strain GBW0011
Separating from natural fermented food, further separating and purifying in laboratory to obtain lactic acid strain with excellent performance, named GBW0011.
As shown in fig. 1 and 2, the colony of strain GBW0011 on the MRS plate is milky to pale yellow in surface, smooth, neat or not very neat in edge, raised in the middle, and opaque; the spores are in a match rod shape under a microscope, the spores are born at the end, some of the spores fall off, some of the spores do not fall off, and the spores are arranged singly.
2. Identification of Strain GBW0011
(1) 16S rRNA Gene identification
The DNA of the strain GBW0011 is used as a template, the 16S rRNA universal primers 27F and 1492R are used for amplification, the amplified fragment is sent to Shanghai workers for sequence determination, the obtained nucleotide sequence is shown as SEQ ID No.1, and the obtained sequence is subjected to BLAST comparison, and the result shows that the homology of the strain GBW0011 and the Weizmannia coagulans sequence in GenBank is 99.18%, so that the strain is preliminarily determined to be Weizhmanni coagulans.
(2) Further characterization of other conserved genes
Appropriate primers are designed by using adk genes (adenylate kinase), recF genes (recombination protein F, recombinant protein F), sucC genes (succinyl-CoA synthetase, sulfokinase) and spo0A genes (Sporulation transcription factor, spore transcription factors), PCR amplification is carried out by taking DNA of a strain GBW0011 as a template, amplified fragments are sent to Shanghai workers for sequence determination, the obtained nucleotide sequences are respectively shown as SEQ ID No.2-SEQ ID No.5, and the sequence is found to have the sequence similarity of 99.23% -100% with Weizmannia coagulans sequences in GenBank through BLAST comparison, as shown in Table 1. Therefore, it was finally confirmed that the strain was Weizhmann's condensation.
Table 1: conservative gene name, fragment size and sequence alignment
3. Preservation of Strain GBW0011
Strain GBW0011 was subjected to strain preservation, collection unit of condensation of GBW0011 of manella wegener: china Center for Type Culture Collection (CCTCC); address: chinese university of Wuhan; preservation date: 2022, 09; the preservation number of the Welch's disease condensation Weizmannia coagulans GBW0011 is CCTCC NO: m20221404.
Example 2: antibacterial effect of Wittman coagulans GBW0011
1. Test method
1. Preparation of cell-free filtrate of Welch's disease
The activated Wettman's bacterium is inoculated in 100mL of fermentation medium (5-8% of bean pulp, 1-2% of peptone, 1-3% of corn flour, 1-1.5% of glucose, 0.3-0.5% of dipotassium hydrogen phosphate, 0.02-0.1% of manganese sulfate and 0.05-0.1% of sodium nitrate) at an inoculation ratio of 2% (v/v), cultured for 24 hours under anaerobic condition, and the obtained culture solution is centrifuged at 5000rpm for 30min, and filtered and sterilized by passing through a 0.22 mu m filter membrane as a supernatant.
2. Recovery and culture of Streptococcus mutans
Thawing the Streptococcus mutans strain stored at-80deg.C, dipping the bacterial liquid in the test tube with sterile cotton swab in an ultra clean bench, uniformly coating on LB solid culture medium plate, and culturing in a 37 deg.C incubator for 16 hr. Picking single colony in solid culture medium with sterile fishing ring, inoculating into 100mL LB liquid culture medium, and collecting strainShake culturing at 37deg.C for 16 hr to logarithmic phase, and regulating bacterial liquid concentration to OD 600 The value is 1.0, and the bacterial content is about 1×10 8 CFU/mL。
3. Recovery and culture of Porphyromonas gingivalis
And taking out the strain stored at the temperature of minus 80 ℃, thawing, dipping the bacterial liquid in a test tube in an ultra-clean bench by using a sterile cotton swab, uniformly coating on a BHI solid culture medium plate, then placing the plate in a constant temperature oven at the temperature of 37 ℃ for anaerobic culture, and observing after 4d of culture. The single colony in the solid culture medium is selected by the sterile bacteria fishing ring and put in a centrifuge tube filled with 10mL BHI liquid culture medium, anaerobic culture is carried out for 3d to logarithmic phase at 37 ℃ in an incubator, and the concentration of the bacterial liquid is regulated to OD 600 The value is 1.0, and the bacterial content is about 1×10 8 CFU/mL。
4. Determination of Minimum Inhibitory Concentration (MIC) of Wittsia coagulans cell-free filtrate
Containing 10 of 8 cfu/mL of fresh culture broth of Streptococcus mutans or Porphyromonas gingivalis was inoculated at a ratio of 1% into 10mL of nutrient broth (concentrations of 0.5%,1.0%,1.5% and 2.0%, respectively) containing 50,100,150,200. Mu.L of the cell-free filtrate of M.coagulans, and cultured at 37℃for 24 hours under aerobic conditions to determine MIC values.
5. Determination of minimum sterilizing concentration (MBC) of Wittman coagulans cell-free filtrate
After the MIC value of the cell-free filtrate of the frozen Wittman bacteria on the strain is measured by an in-vitro bacteriostasis experiment, the cell-free filtrate with the same concentration is mixed with the streptococcus mutans or Porphyromonas gingivalis liquid in a ratio of 1:10, 0.1mL of the mixed filtrate is inoculated on a sterilized LB solid culture medium plate, and then the mixed filtrate is cultured for 24 hours at 37 ℃, and the calculated result is compared with a control treatment group (CFE content is 0%).
2. Test results
Growth of Streptococcus mutans and Porphyromonas gingivalis in the cell-free filtrate of GbW0011 of M.Weizmanni coagulans at different concentrations is shown in Table 2 and FIG. 1, and the inhibition degree of Streptococcus mutans and Porphyromonas gingivalis increases as the concentration of the cell-free filtrate increases after culturing for 24 hours at 37 ℃. Table 3 shows the Minimum Bactericidal Concentration (MBC) of the cell-free filtrate of M.wegener GBW0011 against oral pathogenic bacteria, indicating that S.mutans and Porphyromonas gingivalis can be completely cleared when the concentration of the cell-free filtrate is 1.5%.
MIC and MBC values of the cell-free filtrate of GBW0011 of manella coagulans against streptococcus mutans and porphyromonas gingivalis are shown in table 4: MIC and MBC values of the Wittsia coagulans cell-free filtrate on Streptococcus mutans were 0.5 and 1.0, respectively. When the concentration of the acellular filtrate is 1%, the flat plate does not grow streptococcus mutans, which shows that 1% of the acellular filtrate has a killing effect on streptococcus mutans; MIC and MBC values of the Wittman coagulans cell-free filtrate on Porphyromonas gingivalis were 1.0 and 1.5, respectively. When the concentration of the cell-free filtrate was 1.5%, there was no growth of Porphyromonas gingivalis on the plate, indicating that 1.5% of the cell-free filtrate had a killing effect on Porphyromonas gingivalis.
Table 2: growth of Streptococcus mutans and Porphyromonas gingivalis in different concentrations of cell-free filtrate of M.Weizhimai
(+) macroscopic bacterial growth (turbidity); (-) bacterial growth invisible to the naked eye (clear)
Table 3: growth conditions of streptococcus mutans and porphyromonas gingivalis cultured in different concentrations of the cell-free filtrate of M.Weizhimai
(++) moderate growth; (+) little growth; (-) no growth
* The tube is only inoculated with oral pathogenic bacteria
Table 4: MIC value and MBC value of Wittman coagulans acellular filtrate on oral pathogenic bacteria
Example 3: clinical test of Wittman coagulans GBW0011 against gingival bleeding
1. Preparation of gargle containing Webster coagulans
Inoculating activated Wittman's bacterium to 100mL fermentation medium (5-8% of bean pulp, 1-2% of peptone, 1-3% of corn flour, 1-1.5% of glucose, 0.3-0.5% of dipotassium hydrogen phosphate, 0.02-0.1% of manganese sulfate and 0.05-0.1% of sodium nitrate) at a ratio of 2% (v/v), anaerobic culturing at 37deg.C for 24 hr, collecting bacterial liquid after fermentation, centrifuging at 4deg.C and 2000rpm to remove supernatant, re-suspending with sterile physiological saline, centrifuging to remove supernatant under the same condition, repeating the above operation for 3 times, adding a certain amount of sterile physiological saline to collect bacterial sludge, re-suspending and diluting to bacterial density of 1.0X10 6 CFU/mL was used as a mouthwash containing coagulated Wittman.
2. Clinical test
Study subject inclusion criteria: (1) all patients refer to the relevant standard of the Ministry of health and Press, namely the practical oral science (essence) (3 rd edition) to diagnose the patients, and then confirm that the symptoms of gingival bleeding and halitosis exist; (2) the patient informed and agreed to sign the study informed instructions. Exclusion criteria: (1) gestational and lactating patients; (2) combining systemic diseases; (3) coagulation dysfunction. Patients participating in the trial were divided into a control group (n=30) and a trial group (n=30) according to the random number table method. 13 men and 17 women in the control group; age 25-49 years, average age (37.65 ±5.14); the course of the disease is 1-4 years, and the average course of the disease (2.51+/-0.24) years. Test group, male 16, female 14; age 24-46 years, average age (34.68+ -3.11); the course of the disease is 1-4 years, and the average course of the disease (2.36+/-0.17) years. The general data of gender, age, disease course and the like of the two groups are compared, and the two groups have no obvious difference (P is more than 0.05) and are comparable.
(1) Treatment with compound chlorhexidine collutory in control group: the mouthwash is carried out for 1 day and 2 times in the morning and evening, the mouthwash time is 1min, and the mouthwash can be eaten after 30min intervals, and the administration is continued for 10d. (2) Test group first given compound chlorhexidine gargle treatment: gargling was carried out 1 day 2 times in the morning and evening for a period of 1min, followed by treatment with gargle containing coagulated Wittman's bacteria: the application method comprises taking chlorhexidine collutory at intervals of 30min for 10d. When the toothbrush is used for washing, the liquid is fully contacted with the affected teeth and surrounding soft tissues in the modes of tongue stirring, cheek swelling and the like. Both groups of patients are treated by taking light and digestible food as main raw materials, and are not spicy and greasy, and rechecked after treatment is finished.
(1) Detecting the index: the patient measured gingival crevicular fluid IL-1 beta and TNF-alpha mRNA expression levels before and after treatment, respectively; the patients respectively draw venous blood before and after treatment on an empty stomach, and a semi-automatic blood coagulation analyzer (Chinese Langpen-2048B) is adopted to detect the change of the blood coagulation function; post-treatment observations compare the gingival bleeding improvement in the two groups of patients. Curative effect judgment standard: (1) the effect is shown: after treatment, the bleeding, redness, swelling and the like in the gingival sulcus of the patient completely disappear; (2) the method is effective: after treatment, the number of gingival sulcus bleeding in the patient is significantly reduced, but there is still slight bleeding and redness; (3) invalidation: after treatment, the symptoms of gingival sulcus bleeding, redness and swelling and the like of patients are not improved or even aggravated at all. Total effective rate= (significant effect + effective)/total case number x 100%.
Determination of gingival crevicular fluid IL-1 beta and TNF-alpha mRNA expression levels: extracting total RNA of gingival crevicular fluid sample of patient by using RNA Extraction Kit extraction kit (TakaRa) according to the requirement of the kit, detecting the purity and concentration of the total RNA by using 2.2% formaldehyde denaturing gel electrophoresis and NanaDrop-1000 micro nucleic acid determinator, and absorbance D 260nm /D 280nm >1.80 indicates purity compounding test requirements. cDNA Synthesis reaction reference PrimeScript TM The RT reagent kit was carried out as required by the reverse transcription kit (TakaRa). RT-PCR amplification reaction reference
Premix Ex Taq TM II, the fluorescent quantitative kit (TakaRa) is required to be carried out, and the reaction system is 20uL; cDNA 2uL (1200 ng/uL), upstream and downstream primers 0.8uL (10 umol/L), 2X SYBR Premix Ex Taq II 10uL,RNase free H each 2 O6.4 uL; PCR reaction conditions: pre-denaturation at 95 ℃ for 30s; denaturation at 95℃for 10s, annealing at 60℃for 30s, annealing at 72℃for 10s,44 cycles. Analysis of the dissolved Curve after amplificationThe relative expression levels are shown as the ratio of the target gene to the reference beta-actin, and the immune factor primer sequences are shown in Table 5.
Table 5: real-time quantitative PCR Using primer sequences
(2) Detection result:
the effect of Wittman coagulans in combination with chlorhexidine on the IL-1. Beta. And TNF-alpha mRNA expression levels in gingival crevicular fluid is shown in Table 6: before treatment, the two groups of patients had no significant difference in the expression level of gingival crevicular fluid IL-1 beta and TNF-alpha mRNA (P > 0.05); post-treatment, control and test group patients had significantly lower gingival crevicular fluid IL-1 beta and TNF-alpha mRNA expression levels than pre-treatment (P < 0.05), and post-treatment test group IL-1 beta and TNF-alpha mRNA expression levels significantly lower than control group (P < 0.05).
Table 6: comparison of IL-1 beta and TNF-alpha mRNA expression levels before and after treatment in two groups of patients
The effect of Wittman coagulans in combination with chlorhexidine on venous blood clotting function is shown in Table 7: there was no significant change in clotting function (P > 0.05) before and after treatment in the control group, indicating that chlorhexidine mouthwash had no significant effect on clotting function of venous blood. The prothrombin time, the activated partial thromboplastin time and the thrombin time after treatment of the test group were significantly better than before treatment (P < 0.05), indicating that the combination of wizhima coagulans and chlorhexidine improved venous blood clotting function.
Table 7: comparison of coagulation function before and after treatment of two groups of patients
The effect of Wittman coagulans in combination with chlorhexidine on the therapeutic effect of patients with gingival bleeding is shown in Table 8: the total effective rate of the treatment of the test group is significantly higher than that of the control group, and the difference has statistical significance (P < 0.05).
Table 8: the treatment effect of two groups of gingival bleeding patients is compared
In conclusion, the Wittman's coagulation GBW0011 has obvious capability of inhibiting reproduction of oral pathogenic bacteria, namely streptococcus mutans and porphyromonas gingivalis, and the combination of the bacteria and chlorhexidine can effectively reduce the expression level of gingival crevicular fluid inflammatory factors IL-1 beta and TNF-alpha mRNA of a patient suffering from gingival bleeding, improve venous blood coagulation of the patient suffering from gingival bleeding, and further improve the treatment effect on the patient suffering from gingival bleeding. In addition, when chlorhexidine is used, partial beneficial bacteria can be killed while bacteria inhibition is performed, which is also the root cause of repeated bleeding of gum of a patient, and after chlorhexidine is used, the invention can recover the oral microbiota disorder caused by chlorhexidine by using the condensation Wittman bacterium GBW0011.
The above embodiments are only for illustrating the technical solution of the present invention, and are not limiting; although the invention has been described in detail with reference to the foregoing embodiments, it will be apparent to one skilled in the art that modifications may be made to the technical solutions described in the foregoing embodiments, or equivalents may be substituted for some of the technical features thereof; such modifications and substitutions do not depart from the spirit and scope of the corresponding technical solutions.
Claims (10)
1. A strain of vigilant-condensation GBW0011, characterized in that it is classified and named as vigilant-condensation Weizmannia coagulans, and has a preservation number of CCTCC NO: m20221404.
2. The condensation of wegener 'S disease GBW0011 according to claim 1, wherein the nucleotide sequence of 16S rRNA of the condensation of wegener' S disease GBW0011 is shown in SEQ ID No. 1; the nucleotide sequence of adk is shown in SEQ ID No. 2.
3. Use of GBW0011 of the condensation of mansoni wegener in the preparation of an oral bacteriostatic agent according to claim 1.
4. The use of GBW0011 of w.coagulans according to claim 3 for the preparation of an oral bacteriostat, wherein the oral bacteriostat comprises GBW0011 cell-free filtrate of w.coagulans at a concentration of 0.5% -2.0% v/v.
5. The use of the condensation of the wizmann bacteria GBW0011 in the preparation of an oral bacteriostat according to claim 4, wherein the preparation method of the condensation of the cell-free filtrate of the wizmann bacteria GBW0011 is as follows: the activated Welch mannomyces coagulans is inoculated into a fermentation culture medium at an inoculation ratio of 2-5% v/v, cultured for 24-36 h at 36-38 ℃ under anaerobic condition, and the obtained culture solution is centrifuged and filtered to obtain the supernatant, namely the cell-free filtrate of the Welch mannomyces coagulans GBW0011.
6. The use of GBW0011 of the species weemagglutination as claimed in claim 5 for the preparation of an oral bacteriostat, wherein the fermentation medium has the formula: 5 to 8 percent of soybean meal, 1 to 2 percent of peptone, 1 to 3 percent of corn flour, 1 to 1.5 percent of glucose, 0.3 to 0.5 percent of dipotassium hydrogen phosphate, 0.02 to 0.1 percent of manganese sulfate and 0.05 to 0.1 percent of sodium nitrate.
7. The use of GBW0011 of the condensation of wegener's disease according to claim 3 for the preparation of an oral bacteriostatic agent, wherein the pathogenic bacteria inhibited by said oral bacteriostatic agent are streptococcus mutans and porphyromonas gingivalis.
8. Use of GBW0011 of the condensation of mansoni wegener in the manufacture of a medicament for the prevention and treatment of gingival bleeding according to claim 1.
9. The use of the condensation Wittman's GBW0011 for preparing a medicament for preventing and treating gingival bleeding according to claim 7, wherein the condensation Wittman's GBW0011 can effectively reduce the expression level of gingival crevicular fluid inflammatory factors and improve the coagulation function, thereby achieving the purpose of preventing and treating gingival bleeding.
10. A liquid containing a bacterium, wherein the liquid containing a bacterium comprises at least 1.0X10 as an active ingredient 6 CFU/mL of GBW0011 bacterial solution of M.Weizhengensis and chlorhexidine.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117264806A (en) * | 2023-07-21 | 2023-12-22 | 无锡爱科派生物科技有限公司 | Wettman's bacterium coagulans and application thereof in inhibiting oral pathogenic bacteria |
| CN118345015A (en) * | 2024-06-18 | 2024-07-16 | 青岛诺和诺康生物科技有限公司 | Fermented lactobacillus mucilaginosus and application thereof in preparation of oral health improving products |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117264806A (en) * | 2023-07-21 | 2023-12-22 | 无锡爱科派生物科技有限公司 | Wettman's bacterium coagulans and application thereof in inhibiting oral pathogenic bacteria |
| CN117264806B (en) * | 2023-07-21 | 2024-05-14 | 无锡爱科派生物科技有限公司 | Wettman's bacterium coagulans and application thereof in inhibiting oral pathogenic bacteria |
| CN118345015A (en) * | 2024-06-18 | 2024-07-16 | 青岛诺和诺康生物科技有限公司 | Fermented lactobacillus mucilaginosus and application thereof in preparation of oral health improving products |
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