The Control of Microbial Growth

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1 11/10/2016 PowerPoint Lecture Presentations prepared by Bradley W. Christian, McLennan Community College CHAPTER 7 The Control of Microbial Growth The Terminology of Microbial Control Sepsis refers to bacterial contamination Asepsis is the absence of significant contamination Aseptic surgery techniques prevent the microbial contamination of wounds 1

2 The Terminology of Microbial Control Sterilization: removing and destroying all microbial life Commercial sterilization: killing C. botulinum endospores from canned goods Disinfection: destroying harmful microorganisms Antisepsis: destroying harmful microorganisms from living tissue The Terminology of Microbial Control Degerming: the mechanical removal of microbes from a limited area Sanitization: lowering microbial counts on eating utensils to safe levels Biocide (germicide): treatments that kill microbes Bacteriostasis: inhibiting, not killing, microbes 2

3 The Rate of Microbial Death Learning Objective 7-2 Describe the patterns of microbial death caused by treatments with microbial control agents. Figure 7.1a Understanding the Microbial Death Curve. 3

4 Physical Methods of Microbial Control Learning Objectives 7-4 Compare the effectiveness of moist heat (boiling, autoclaving, pasteurization) and dry heat. 7-5 Describe how filtration, low temperatures, high pressure, desiccation, and osmotic pressure suppress microbial growth. 7-6 Explain how radiation kills cells. Moist Heat Sterilization Moist heat denatures proteins Boiling Free-flowing steam 4

5 Moist Heat Sterilization Autoclave: steam under pressure 121 C at 15 psi for 15 min Kills all organisms and endospores Steam must contact the item's surface Figure 7.2 An autoclave. Exhaust valve (removes steam Steam to after sterilization) chamber Safety valve Pressure gauge Operating valve (controls steam from jacket to chamber) Steam Door Steam chamber Air Perforated shelf Steam jacket Sediment screen Thermometer To waste line Automatic ejector valve (thermostatically controlled; closes on contact with pure steam when air is exhausted) Steam supply Pressure regulator for steam supply 5

6 Moist Heat Sterilization Large containers require longer sterilization times Test strips are used to indicate sterility Table 7.4 The Effect of Container Size on Autoclave Sterilization Times for Liquid Solutions* 6

7 Figure 7.3 Examples of sterilization indicators. Heat Pasteurization reduces spoilage organisms and pathogens Equivalent treatments 63 C for 30 min High-temperature short-time (HTST): 72 C for 15 sec Ultra-high-temperature (UHT): 140 C for 4 sec Thermoduric organisms survive 7

8 Dry Heat Sterilization Kills by oxidation Flaming (liekitys) Incineration (poltto) Hot-air sterilization Filtration Passage of substance through a screenlike material Used for heat-sensitive materials High-efficiency particulate air (HEPA) filters remove microbes >0.3 µm Membrane filters remove microbes >0.22 µm 8

9 Figure 7.4 Filter sterilization with a disposable, presterilized plastic unit. Flask of sample Cap Membrane filter Cotton plug in vacuum line ensures sterility Sterile filtrate Vacuum line Physical Methods of Microbial Control Low temperature has a bacteriostatic effect Refrigeration Deep-freezing Lyophilization (freeze drying) High pressure denatures proteins Desiccation (kuivatus): absence of water prevents metabolism Osmotic pressure uses salts and sugars to create hypertonic environment; causes plasmolysis 9

10 Radiation Ionizing radiation (X rays, gamma rays, electron beams) Ionizes water to create reactive hydroxyl radicals Damages DNA by causing lethal mutations Nonionizing radiation (UV, 260 nm) Damages DNA by creating thymine dimers Microwaves kill by heat; not especially antimicrobial Figure 7.5 The radiant energy spectrum nm 10 3 nm 1 nm 10 3 nm 10 6 nm 1 m (10 9 nm) 10 3 m Gamma rays X rays UV Infrared Microwaves Radio waves Bactericidal Tanning Ultraviolet (UV) light UV in sunlight Visible light 200 nm 250 nm 300 nm 350 nm 400 nm 450 nm 500 nm 550 nm 600 nm 650 nm 700 nm 750 nm 280 nm 295 nm 330 nm Wavelength increases Energy increases 10

11 Alcohols Denature proteins and dissolves lipids No effect on endospores and nonenveloped viruses Ethanol and isopropanol Require water Table 7.6 Biocidal Action of Various Concentrations of Ethanol in Aqueous Solution against Streptococcus pyogenes 11

12 Heavy Metals and Their Compounds Oligodynamic action very small amounts exert antimicrobial activity Denature proteins Ag, Hg, Cu, Zn Silver nitrate is used to prevent ophthalmia neonatorum (vastasyntyneiden silmätulehdus) Mercuric chloride prevents mildew (homesienityyppi) in paint Copper sulfate is an algicide Zinc chloride is found in mouthwash Figure 7.8 Oligodynamic action of heavy metals. 12

13 Figure 7.10 A comparison of the effectiveness of various antiseptics. The Disk-Diffusion Method Evaluates efficacy of chemical agents Filter paper disks are soaked in a chemical and placed on a culture Look for zone of inhibition around disks Millaisia kemikaaleja voi testata? What kinds of chemical substances can be tested? 13

14 Figure 7.6 Evaluation of disinfectants by the disk-diffusion method. Zone of inhibition Chlorine Chlorine Chlorine O-phenylphenol Hexachlorophene Hexachlorophene O-phenylphenol Hexachlorophene O-phenylphenol Quat Quat Quat Staphylococcus aureus (gram-positive) Escherichia coli (gram-negative) Pseudomonas aeruginosa (gram-negative) 14

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