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Chapter 5.9 : Agar Dilution MIC Test for Anaerobic Bacteria

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Abstract:

The anaerobe agar dilution MIC method is a semiquantitative method for the determination of in vitro activity of an antimicrobial agent against anaerobic bacterial isolates. A series of agar plates, each containing a unique concentration of an antimicrobial agent, are inoculated with up to 36 isolates. After 48 h of incubation at 35°C in an anaerobic environment, the MIC is determined by observing the lowest concentration of antimicrobial agent which will inhibit visible growth of the test isolate. MICs obtained are interpreted as susceptible, intermediate, or resistant based on criteria defined by the NCCLS ( ).

Citation: Garcia L. 2010. Agar Dilution MIC Test for Anaerobic Bacteria, p 77-88. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch5.9
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Anaerobe Agar Dilution MIC QC

Citation: Garcia L. 2010. Agar Dilution MIC Test for Anaerobic Bacteria, p 77-88. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch5.9
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References

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1. Jousimies-Somer, H. R.,, P. Summanen,, D. M. Citron,, E. J. Baron,, H. M. Wexler,, and S. M. Finegold. 2002. Wadsworth-KTL Anaerobic Bacteriology Manual, 6th ed. Star Publishing Co., Belmont, Calif.
2.NCCLS. 2001. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Anaerobically, 5th ed.Approved standard M11-A5. NCCLS, Wayne, Pa.
3. Citron, D. M.,, and D. W. Hecht,. 2003. Susceptibility test methods: anaerobic bacteria, p. 11411148. In P. R. Murray,, E. J. Baron,, J. H. Jorgensen,, M. A. Pfaller,, and R. H. Yolken(ed.), Manual of Clinical Microbiology, 8th ed. ASM Press, Washington, D.C.
4. Finegold, S. M.,, and W. L. George. 1989. Anaerobic Infections in Humans. Academic Press, Inc., San Diego, Calif.
5. Holdeman, L. V.,, E. P. Cato,, and W. E. C. Moore (ed.). 1977>. Anaerobe Laboratory Manual, 4th ed. Virginia Polytechnic Institute and State University, Blacksburg.
1. Jousimies-Somer, H. R.,, P. Summanen,, D. M. Citron,, E. J. Baron,, H. M. Wexler,, and S. M. Finegold. 2002. Wadsworth-KTL Anaerobic Bacteriology Manual,6th ed. Star Publishing Co., Belmont, Calif.
2.NCCLS. 2001. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Anaerobically, 5th ed. Approved standard M11-A5. NCCLS, Wayne, Pa.

Tables

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Citation: Garcia L. 2010. Agar Dilution MIC Test for Anaerobic Bacteria, p 77-88. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch5.9
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Preparation of antimicrobial dilutions from stock solutions

Reproduced with permission from 2003. Thirteenth informational supplement M100-S13. NCCLS, Wayne, Pa. Copies of the current edition may be obtained from NCCLS, 940 West Valley Rd., Suite 1400, Wayne, PA 19087-1898. also 2001. 5th ed. Approved standard M11-A5. NCCLS, Wayne, Pa. The antimicrobial agent is diluted 10-fold when it is added to the agar. Therefore, the intermediate antimicrobial concentration is 10 times the final concentration of the antimicrobial agent in the agar.

Begin with stock solution at a concentration of 5,120 or 2,000 μg/ml.

Citation: Garcia L. 2010. Agar Dilution MIC Test for Anaerobic Bacteria, p 77-88. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch5.9
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Volumes of components required for preparation of agar dilution plates when using round or square petri plates

Citation: Garcia L. 2010. Agar Dilution MIC Test for Anaerobic Bacteria, p 77-88. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch5.9
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Patterns for arrangement of isolates in Steers replicator

Citation: Garcia L. 2010. Agar Dilution MIC Test for Anaerobic Bacteria, p 77-88. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch5.9
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Agar Dilution MIC Worksheet

Record growth (+ or -) for each organism (each drug concentration and growth control plates).

*Pre: growth control plates inoculated immediately preceding first dilution of antimicrobial-containing plates

**Post: growth control plates inoculated immediately following last dilution of antimicrobial-containing plates

Citation: Garcia L. 2010. Agar Dilution MIC Test for Anaerobic Bacteria, p 77-88. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch5.9

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