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Microbio Lab 5

Microbio Lab 5

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Published by: nkivc on Nov 16, 2009
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MICROBIOLOGY LABORATORY 5USTMED ’07 Sec C – AsM; Photos provided by JV.N. & MeaM.SENSITIVITY TESTINGAntibiotic Sensitivity Testing Using The Kirby-BauerProcedure
Kirby Bauer method. The test, introduced by William Kirby andAlfred Bauer in 1966, consists of exposing a newly-seeded lawnof the bacterium to be tested, growing on a nutrient medium(Mueller-Hinton agar) to filter paper disks impregnated withvarious antibiotics. The culture is incubated for 16 to 18 hoursand then examined for growth. If the organism is inhibited byone of the antibiotics, there will be a zone of inhibition aroundthe disk, representing the area in which the organism wasinhibited by that antibiotic.The diameter of the zone of inhibition around an antibiotic diskis an indication of the sensitivity of the tested microorganismto that antibiotic. The diameter of the zone, however, is alsorelated to the rate of diffusion of the antibiotic in the medium.This fact must be kept in mind then interpreting the zone of inhibition of various antibiotics.MATERIALS:-culture of the organisms in Mueller-Hinton agar platewith antibiotic sensitivity discs-Ruler graduated in millimetersNOTE:-The complete procedure is in you(r) lab [lab u 2!Hahaha!] manual.1.Measure the zone of inhibition and record yourresults.2.Interpret the results based on the table provided
Єtest – The Problem Solver in AntimicrobialSusceptibility Testing
Etest is an antimicrobialgradient strip for thequantitative determinationof susceptibility orresistance omicroorganisms. It is arobust and simpletechnique, minimallyaffected by laboratoryvariations and can be usedto test mostmicroorganisms. Anaccurate and reproducibleMinimum InhibitoryConcentration (MIC) isgenerated for reliableguidance of antimicrobialtherapy. 
Єtest Susceptibility Testing
inoculation of theorganism for testing usingcotton swab by overlappingstreaking using MuellerHinton Agar Plate.2.Overlaying of Єteststrip on the previouslyinoculated culture media3.Reading of the strip and recording the results
Disk Diffusion Method 
Procedure1.dip the sterile swab into bacterial suspensioncompared to 0.5 MF standard then swab onto thesurface of Mueller Hinton Agar using Overlappingtechnique.
Allow the organism to be absorbed by the medium.Place the appropriate antimicrobial (sensitivity) discsusing the dispenser or a sterile forceps. Incubate for24 hours at 37
3.Reading and Interpretation of the results. Measurethe diameter of the Zone of Inhibition (area whereinthere is no growth around the discs) using themillimeter of a ruler. Record your results andInterpret based on the table provided. Determine if the Antibiotic (organism) is Sensitive, Intermediate orResistant. If there is overlapping in the zone of inhibition, you can just measure the radius andmultiply the reading by 2 to get the diameter.E. coli ATCC 25922. Theisolate tested on this Mueller-Hinton agar plate isinterpreted as susceptible (S)to all antimicrobial agents.Reading clockwise from thetop, MZ, AN, AM, CZ, CTX,CXM, CF, GM, NN: the threediscs in the center of theplate are SXT, FOX and TIM.Pseudomonas aeruginosa, aresistant strain. Growth onthis Mueller-Hinton agar plateindicates that the isolate isresistant to six of 12antimicrobial agents andsusceptible to the remaing.The isolate is resistant toSXT, GM, ATM, TIM, TIC andMMZ. The isolate issusceptible to CIP, AN, NN,CA, IPM and PIP.u stmedc3@yahoogroups.coma udrey_cl@yahoo.com- fin -

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