Method for reducing the transfer of pathogenic microorganisms
A technology of pathogenic microorganisms and microorganisms, applied in the direction of microorganism-based methods, microorganisms, microorganisms, etc., can solve problems such as the difficulty in determining the source of MRSA infection strains
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Embodiment 1
[0233] Example 1 strain screening and identification
[0234] sample
[0235] In order to identify and select bacterial strains according to the invention, a strain collection of lactic acid bacteria (LAB) was established. Samples from different sources such as homemade sauerkraut, kimchi and healthy human donor samples (vaginal, oral, anal, skin) were collected to isolate at least 995 species of lactic acid bacteria. Samples were collected in Man Rogosa Sharp (MRS, Sigma-Aldrich) broth and agar and incubated anaerobically at 37°C overnight or until colonies formed. Isolates were plated and subcultured until pure colonies were obtained. Pure colonies were stored in MRS broth with 25% glycerol at -80 °C for future use. Strains were identified using standard methods of 16SrRNS Sanger sequencing.
Embodiment 2
[0236] Example 2: Co-culture Assay / Competition Assay
[0237] Competition between bacterial strains and Staphylococcus aureus was identified (Dowarah, R., et al. 2018, Selection and characterization of probiotic lactic acid bacteria and its impact on growth, nutrient digestibility, health and antioxidant status inweaned piglets.PLoS ONE, 13(3), Khare, A., & Tavazoie, S. (2015). Multifactorial Competition and Resistance in a Two-Species Bacterial System. PLoS Genetics, 11(12), 1–21.).
[0238] S. aureus subsp. aureus COL (CCOS461), S. aureus CC398 (SSI strain 08S00974) and S. aureus US300 (ATCC BAA-1717) were used as MRSA test organisms. Staphylococcus aureus was cultured in brain heart infusion (BHI) broth.
[0239] Overnight cultures of S. aureus and isolates were adjusted to an optical density at 600 nm (OD600) of 1 and harvested by centrifugation (6.000 rpm for 2 minutes). Cell pellets were washed twice in phosphate buffered saline (1×PBS) and then resuspended in 1×PBS. ...
Embodiment 3
[0241] Example 3: Determining transfers between objects
[0242]The transfer of pathogenic bacteria between two subjects was determined by a co-culture assay. Overnight cultures of Staphylococcus aureus and bacterial strain isolates were adjusted for cell density to an optical density at 600 nm (OD600) of 1 and harvested by centrifugation (6.000 rpm for 2 minutes). Wash the cell pellet twice in phosphate-buffered saline (1×PBS), and resuspend in 1×PBS to make the cell concentration approximately 1×10 4 CFU / ml. The two cell suspensions were mixed 1:1, and 0.2 ml was spread on a BHI agar plate and allowed to dry in sterile air for 5 minutes. Monocultures of MRSA and LAB were used as controls, and for control plates, the cell suspension was mixed 1:1 with sterile PBS buffer, 0.2 ml was spread on BHI agar plates, and allowed to dry in sterile air for 5 min .
[0243] 5 ml sterile PBS was absorbed through a sterile round filter (D 9 cm) type A.G.F 118 and placed on top of the a...
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