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Citrex 2

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Citrex 2

  1. 1. MINIMUM INHIBITORY CONCENTRATION TEST : MIC TEST
  2. 2. MATERIALS AND METHODS <ul><li>Microorganisms </li></ul><ul><li>Concentration of BIO-CLEANTM </li></ul><ul><li>Procedure and assay </li></ul>
  3. 3. MICROORGANISMS <ul><li>The American Type Culture Collection strain [ATCC] </li></ul><ul><li>Department of Medical Sciences Thailand strain [DMST] </li></ul><ul><li>Escherichia coli ATCC 25922 </li></ul><ul><li>Salmonella Typhimurium DMST 15674 </li></ul><ul><li>Salmonella Enteritidis DMST 15676 </li></ul><ul><li>Enterococcus faecium DMST 17660 </li></ul><ul><li>Enterococcus faecalis ATCC 29212 </li></ul>
  4. 4. MICROORGANISMS (CONT.) <ul><li>Enterococcus faecalis ATCC 51299 </li></ul><ul><li>Enterobacter aerogenes DMST 13048 </li></ul><ul><li>Bacillus cereus ATCC 11778 </li></ul><ul><li>Pseudomonas aeruginosa ATCC 27853 </li></ul>
  5. 5. CONCENTRATION OF BIO-CLEANTM <ul><li>The following BIO-CLEANTM concentrations were tested : </li></ul><ul><li>1,000 ppm </li></ul><ul><li>900 ppm </li></ul><ul><li>800 ppm </li></ul><ul><li>700 ppm </li></ul><ul><li>600 ppm </li></ul><ul><li>500 ppm </li></ul><ul><li>400 ppm </li></ul><ul><li>300 ppm </li></ul><ul><li>200 ppm </li></ul><ul><li>100 ppm </li></ul>
  6. 6. PROCEDURE AND ASSAY <ul><li>Preparation of microorganisms </li></ul><ul><li>Cultivated microorganisms in Tryptic soy broth (TSB) and incubated 37 ° C at 24 hours. </li></ul><ul><li>Microorganisms suspension to McFarland’s pattern of 105 cfu/ml was prepared. </li></ul>
  7. 7. FLOW CHART (PREPARATION OF MICROORGANISM) Compared with 0.5 McFarland = 10 5 cfu/ml ten fold dilution with PSS for Mcfarland Culture in TSB medium Slant agar Streak on NA agar and incubated at 37 o C; 24 hrs Picked 3 colonies 10 -1 10 -2 10 -3 10 -4 10 -5 10 -6 10 -7 10 -8 Incubated at 37 o C ; 24 hrs
  8. 8. PROCEDURE AND ASSAY (CONT.) <ul><li>Preparation of Mcfarland 105 cfu/ml </li></ul><ul><li>To standardized the inoculum density is used (0.5 McFarland standard) </li></ul>
  9. 9. PREPARATION OF 0.5 MCFARLAND
  10. 10. PREPARATION OF 0.5 MCFARLAND
  11. 11. FLOW CHART (PREPARATION OF 0.5 MCFARLAND) OD 625 = 0.8-0.10 0.5 McFarland BaCl 2 + H 2 SO 4 (Dilution) 1:1 1:2 1:3 1:4 1:5
  12. 12. PROCEDURE AND ASSAY (CONT.) <ul><li>Controls </li></ul><ul><li>Control test tube 1 = 1 ml TSB + 1 ml microorganism </li></ul><ul><li>Control test tube 2 = 1 ml TSB + 1 ml BIO-CLEANTM </li></ul><ul><li>Control test tube 3 = 2 ml TSB </li></ul>C1 C2 C3
  13. 13. PROCEDURE AND ASSAY (CONT.) <ul><li>Sample </li></ul><ul><li>1 ml BIO-CLEANTM (start 1,000-100ppm) </li></ul><ul><li>1 ml microorganism (105 cfu/ml) </li></ul>1,000 ppm
  14. 14. PROCEDURE AND ASSAY (CONT.) --- -+- -++ Report C1 C2 C3 1,000 ppm 900 ppm 800 ppm 700 ppm 600 ppm 500 ppm 400 ppm 300 ppm 200 ppm 100 ppm

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