Chapter 7 Reading Worksheet

Name ___________________________

Define the following terms:

1. Sterilization
- Removal of all microorganisms, including endospores
2. Disinfection
- Any treatment used on inanimate objects to kill or inhibit the growth of
microorganism
- A chemical used is called a disinfectant
3. Antisepsis
- A chemical method for disinfection of the skin or membranes
- The chemical is called an antiseptic
What is the primary difference between the terms disinfection and antisepsis?
- Whether the treatment is being directed at living tissue
PHYSICAL METHODS OF MICROBIAL CONTROL
4. How does moist heat kill microorganisms?
- Primarily by coagulating proteins/enzymes (denaturation), caused by breakage of
hydrogen bonds that hold the proteins in their three-dimensional structure
5. What is the temperature/pressure/time requirement for sterilization using a steam
autoclave (at sea level)?
- 121C
- 15 psi
- 15 minutes
6. How does the sterilization protocol change as the volume of the material being
autoclaved changes?
- Sterilization time increases as volume of material increases
7. a. Define thermoduric and give two examples of thermoduric bacteria.
- Thermoduric: heat resistant
- Bacillus
- Clostridium
- Enterococci
b. Differentiate the terms thermophilic and thermoduric.
- Thermophilic: optimum temperature of about 60C

- Growth temperatures range from 45-85C

- Thermoduric: can survive at extreme temperatures, like pasteurization, but
cannot grow
8. List the time/temp. requirements for the following:
Traditional classic (or batch) pasteurization (try the Internet): 30 min/63 C
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High-temperature short time pasteurization (HTST): 15 sec/72C

Do either of these methods sterilize the material being pasteurized?
- No
What are the two primary goals of pasteurization?
a. Destroy/remove pathogenic microorganisms
b. Extend shelf life by reducing spoilage organisms
What is UHT? Name a product that this is used on.
- Ultra-high-temperature Treatment
- Used on dairy products can be stored in sealed containers at room
temperature
What is accomplished by UHT?
- Sterilizes the product
9. Provide two examples of applications of dry heat sterilization?
- Direct flaming or incineration
- Hot-air sterilization
10. List time/temp. for hot air sterilization:
- 2 hours/170C
11. What is a HEPA filter? How might these be used in a medical application?
- High-efficiency particulate air filter: screen-like material that removes particles
larger than 0.3 m from air
12. What are membrane filters and what kind of materials might they be used to
sterilize?
- Screen-like material with pores small enough to retain microorganisms
- 0.45 m filter retains most bacteria
- Appropriate for heat labile liquids
13. Describe desiccation and how this controls microbial growth:
- Water removal prevents metabolism
- Organisms cannot grow or reproduce but remain viable for years
- When water made available to them resume their growth and division
14. How might osmotic pressure be used to inhibit bacterial growth?
- High concentrations of salts and sugars create hypertonic environment that
causes water to leave microbial cell
- Denies cell moisture needed for growth
14. What is ionizing radiation and how does it damage bacteria?
- High-energy radiation with a wavelength less than 1nm causes ionization
- X rays and gamma rays
- Ionization of water forms highly reactive hydroxyl radicals
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- Radicals kill organisms by reacting with organic cellular components and

damaging them
What materials might be sterilized using ionizing radiation?
- Pharmaceuticals and disposable dental and medical supplies
- Plastic syringes, surgical gloves, suturing materials, catheters
- Sterilization of certain classes of mail by postal service
Is ionizing radiation ever used on foods? (check out p. 797-798)
- Low doses: kill insects (disinfestation) and inhibit sprouting, delay ripening
of fruits during storage
- Pasteurizing doses: used on meats and poultry to eliminate or critically
reduce numbers of specific bacterial pathogens
- High doses: sterilize or greatly lower bacterial populations in many spices
15. How does UV light damage bacterial cells?
- Damages DNA of exposed cells by causing bonds to form between pyrimidine
bases, usually thymines, in DNA chains
- Thymine dimers inhibit correct replication of the DNA during cell reproduction
What are some practical situations in which UV sterilization or disinfection is used?
- Control microbes in the air hospital rooms, nurseries, operating rooms,
cafeterias
- Disinfect vaccines and other medical products
What is the major problem/consideration with using UV sterilization?
- Non-penetrating: cant pass through paper, liquid, plastic, glass
- Harmful to eyes and skin
- Prolonged exposure can cause burns and cancer in humans
CHEMICAL METHODS OF MICROBIAL CONTROL
What are two ways that chemical agents can be evaluated?
a. Use-dilution test: method of determining effectiveness of disinfectant using serial
dilutions
b. Disk-diffusion method: agar-diffusion test to determine microbial susceptibility to
chemotherapeutic agents
Provide the indicated information for each of the chemical agents listed.
Phenol and Phenolics:
Phenol
Disadvantage:
- Irritates skin and has disagreeable odor
Historical interest:
- First used by Joseph Lister to control surgical infections in the
operating room
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Phenolics
How do they differ from phenol?
- Contain molecule of phenol that has been chemically altered to
reduce its irritating qualities or increase its antibacterial activity in combination with a
soap or detergent
Advantages:
- They remain active in presence of organic compounds, are stable,
persist over long periods after application
Example:
- Lysol
How might a phenolic (cresol) be used?
- As a surface disinfectant
Bisphenol
Example:
- Prescription lotion, pHisoHex, used for surgical and hospital microbial
control procedures
- Triclosan
- Antimicrobial soap
- Toothpaste
- Kitchen cutting boards and handles of knives and other plastic
kitchenware
What group of organisms is it most effective against?
- Gram-positive staphylococci and streptococci
Common use of this agent:
- Plastic: food packaging and storage
Bioguanides:
Chlorhexidine/Alexidine
Advantages:
- Primarily affect bacterial cell membranes
- Particularly effective against gram-positive bacteria
- Also effective against gram-negative bacteria with
exception of pseudomonads
- Not sporicidal but have some activity against enveloped viruses
Common use of this agent:
- Frequently used for microbial control on skin and mucus membranes
- Combined with detergent or alcohol, chlorhexidine often used for
surgical hand scrubs and preoperative skin preparation in patients
- Alexidine is similar and more rapid in action vs. chlorhexidine
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- PerioGard oral rinse

- Hibiclens
Halogens (chlorine and iodine containing compounds)
Iodophors
Define iodophor:
- Complex of iodine and a detergent
- Iodine bound to organic molecule
Common commercial preparation:
- Antiseptic
Advantages:
- Antimicrobial activity of iodine, but do not stain and are less irritating
Common applications of this compound:
- Betadine: providone-iodine
- Improves wetting action and serves as reservoir of free iodine
- Iodines used mainly for skin disinfection and wound treatment
Chlorine compounds
Sodium hypochlorite:
Common name:
- Clorox
Common uses:
- Household disinfectant and bleach
- Disinfectant in dairies, food-processing establishments,
hemodialysis systems
Particularly useful for killing (inactivating) bacteria, fungi, and viruses.
Calcium hypochlorite:
- Disinfect dairy equipment and restaurant eating utensils
Chlorine dioxide:
- Surface disinfectant in food-processing industry
- Sewage water disinfection
- Industrial air treatment
- Fumigate enclosed building areas contaminated with endospores of
anthrax
- Antiseptic at low concentrations
- Gas sterilization of medical equipment
Chloramines:
- Chlorine + ammonia
- Municipal water-treatment systems
- Sanitize glassware and eating utensils
- Treat dairy and food-manufacturing equipment

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Alcohols
Preferred type of alcohol for antisepsis and disinfection:
- Isopropanol
Why is 70% alcohol preferred over 100%?
- Less effective than aqueous solutions because denaturation requires water
What is meant by the term tincture?
- A solution in aqueous alcohol
Heavy Metals
Define: Oligodynamic action:
- The ability of small amounts of a heavy metal compound to exert
antimicrobial activity
Silver nitrate
Historical use:
- Used as antiseptic
- Many states used to require that eyes of a newborn be treated with a
few drops of silver nitrate to guard against infection
Silver sulfadiazine
How and why is this used?
- As a topical cream for burns to use silver ions against bacteria
What is the metal in X-gel and what is X-gel used for?
- Copper is used as a hand sanitizer

Surface Active Agents

Soaps and Detergents:
- Functions in mechanical removal of microbes through scrubbing
- Soap breaks oily film on skin into tiny droplets (emulsification) and water and
soap together lift up emulsified oil and debris and float them away as lather is washed
off
Acid-anionic Sanitizers:
- Important in cleaning food-processing facilities
- Usually combinations of phosphoric acid with a surface-active agent
Quaternary Ammonium Compounds (Quats)
Examples of products that contain these (3):
- Zephiran
- Cepacol
- Mouthwash
- Clorox
- Lysol
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- Cavicide
Common uses of products that contain quats:
- Bactericidal, fungicidal, amebicidal, virucidal
Notable bacterial species that is NOT killed by quats:
- Pseudomonas
Aldehydes
Glutaraldehyde/Ortho-phthalaldehyde (OPA)
Common commercial preparation of 2% glutaraldehyde:
- Cidex
Common uses of glutaraldehyde:
- Disinfect hospital instruments including endoscopes and respiratory
therapy equipment
Does glutaraldehyde sterilize?
- Yes
Ethylene Oxide
How is this used?
- Closed chamber similar to a steam autoclave
Does it sterilize?
- Yes
Advantages:
- Carries out sterilization at ambient temperatures and is highly penetrating
Disadvantages:
- Lengthy exposure period
- Toxic and explosive in its pure form
Plasmas
What is plasma?
- Excited gases used for sterilizing
What is plasma sterilization particularly useful for?
- Tubular instruments/endoscopy
equipment
- Items that are temperature or moisture sensitive
What chemical is used?
- Hydrogen peroxide activated by vacuum and electromagnetic field to form
plasma gas
- Free radicals destroy microbes
Supercritical Fluids
Briefly describe:
- CO2 compressed into a supercritical state, with gaseous (lowered surface
tension) and liquid (increased solubility) properties
What have supercritical fluids been used for?
- Tissue medical implants (bone, tendons, ligaments) taken from donors
Peroxygens
Hydrogen peroxide
Why is H2O2 not a good antiseptic agent for open wounds?
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- Quickly broken down to water and gaseous oxygen by action of

enzyme catalase, present in human cells
What are some appropriate applications of H2O2 for disinfection?
- Inanimate objects
- Contact lenses
How is Bioquell used?
- Hot gaseous H2O2 used for sterilization of atmosphere and surfaces
including hospital rooms
Peracetic acid
List several appropriate uses for this compound:
- Washing fruits and vegetables
- Food processing and medical equipment
Advantages:
- Effective on endospores and viruses within 30 minutes and kills
vegetative bacteria and fungi in less than 5 minutes
- Leaves no toxic residues
Summary:
List 3 chemical sterilants:
1. Ethylene oxide
2. Chlorine oxide
3. Peracetic acid
List 2 common products used as surgical scrubs for skin antisepsis skin prior to
surgery:
1. Chlorhexidine
2. Betadine
List several methods used to improve food safety:

List 3 microorganisms or microbial forms that are a major challenge to disinfection and
antisepsis:
1. Pseudomonas
2. Clostridium difficile
3. Mycobacteria
Prions have provided a major challenge to routine sterilization protocols. What methods
are available for prion inactivation/destruction? (do an Internet search for this one)
- Autoclaving at 121C for 15 mins
- Boiling in sodium dodecyl sulfate (3%)
- Iodophores
- Chlorine dioxide
- Glutaraldehyde

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