2120 Lab. Week 11. Experiments 13,14,21. Kirby Bauer, TDT, Chemicals

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1 2120 Lab Week 11 Experiments 13,14,21 Kirby Bauer, TDT, Chemicals

2 Controlling Microorganisms Decontamination: Physical, chemical, and mechanical methods to destroy or reduce undesirable microbes in a given area Primary targets are microorganisms capable of causing infection or spoilage: Vegetative bacterial cells and endospores Fungal hyphae and spores, yeast Protozoan trophozoites and cysts Worms Viruses Prions

3 Microbial Control Terminology

4 The Resistance of Endospores

5 1) Number of microbes Factors That Affect Death Rate 2) Nature of microbes in the population 3) Temperature and ph of environment 4) Concentration or dosage of agent 5) Mode of action of the agent 6) Presence of solvents, organic matter, or inhibitors

6 1. Heat moist and dry Methods of Physical Control 2. Cold temperatures 3. Desiccation 4. Radiation 5. Filtration

7 Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Mode of Action and Relative Effectiveness of Heat Moist heat lower temperatures and shorter exposure time; coagulation and denaturation of proteins Number of cells added to population (growth) Number of Cells that survive Dry heat moderate to high temperatures; dehydration, alters protein structure; incineration Temperature C

8 Comparing Moist Heat to Dry Heat

9 Thermal Death Measurements Bacterial endospores most resistant usually require temperatures above boiling Thermal death time (TDT) shortest length of time required to kill all test microbes at a specified temperature Thermal death point (TDP) lowest temperature required to kill all microbes in a sample in 10 minutes

10 Boiling Water Boiling at 100 o C for 30 minutes to destroy non-spore-forming pathogens Disinfection

11 Relative Resistance of Microbes Highest resistance Prions, bacterial endospores Moderate resistance Pseudomonas sp. Mycobacterium tuberculosis Staphylococcus aureus Protozoan cysts Least resistance Most bacterial vegetative cells Fungal spores and hyphae, yeast Enveloped viruses Protozoan trophozoites

12 Chemical Agents in Microbial Control Disinfectants, antiseptics, sterilants, degermers, and preservatives Some desirable qualities of chemicals: Rapid action in low concentration Solubility in water or alcohol, stable Broad spectrum, low toxicity Penetrating Noncorrosive and non-staining Affordable and readily available

13 Chemical Agents Used in Health Care

14 The Active Ingredients of Various Commercial Antimicrobial Products

15 Terminology

16 Identifying the infectious agent Identification of infectious agent should be attempted as soon as possible. Specimens should be taken before antimicrobials are initiated.

17 Testing for Drug Susceptibility Kirby-Bauer disk diffusion test Medium used: Mueller- Hinton agar OT 30 0mm ENR GN 10 1) Measure radius 2) Multiply by 2 CTX 30 AMP 10 C 30 Measure diameter AMP 5 Antibiotic carrier (disc)imprinted with abbreviation and concentration Zone of Inhibition Region of bacterial growth

18 Comparing MICs for Common Drugs and Pathogens Minimum inhibitory concentration (MIC) smallest concentration of drug that visibly inhibits growth

19 Testing for Drug Susceptibility Essential for groups of bacteria commonly showing resistance Kirby-Bauer disk diffusion test E-test diffusion test Minimum inhibitory concentration (MIC) smallest concentration of drug that visibly inhibits growth (Dilution test) Same inoculum size of test bacteria added Negative mg/ml control Increasing concentration of drug Growth No growth Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

20 The MIC and Therapeutic Index In vitro activity of a drug is not always correlated with in vivo effect If therapy fails, a different drug, combination of drugs, or different administration must be considered Best to chose a drug with highest level of selectivity but lowest level toxicity measured by therapeutic index Therapeutic index: the ratio of the dose of the drug that is toxic to humans as compared to its minimum effective dose High index is desirable Toxic dose= 200 ug/ml TI= 2 MIC = 100 ug/ml Toxic dose= 10 ug/ml TI= 0.1 MIC = 100 ug/ml

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