Mariela Cassandra L.

Mendoza
MBB 110: Exercise 1
STERILIZATION AND THE ASEPTIC TECHNIQUE
1. Define sterilization, disinfection, and decontamination. What are their differences and when are
they performed?
Sterilization is a set of procedures done to eliminate all forms of life, mainly microorganisms from
any material particularly glassware, equipment and solutions to be used in the laboratory. Heat,
ethylene oxide gas, hydrogen peroxide gas, plasma, ozone and radiation are used in sterilization
procedures. The sterility assurance level is attained if the probability of a microorganism
surviving on an item subjected to the procedure is less than one in one million (Chosewood &
Wilson, 2009).
Disinfection is a process wherein all recognized pathogenic microorganisms are eliminated from
an item. Unlike sterilization, disinfection does not necessarily eliminate all forms of microbial life
(e.g. bacterial spores) (Chosewood & Wilson, 2009). The effectiveness of a disinfection procedure is
dependent on factors such as the nature and number of contaminating microorganisms (especially
the presence of bacterial spores), amount of organic matter present (e.g., soil, feces and blood), type
and condition of materials to be disinfected and the temperature. Chosewood and Wilson (2009)
also stated that disinfection procedures are classified into three levels in terms of the effectiveness
of the procedure in reducing the level of microbial contamination in an object:
High-level disinfection process eliminates vegetative microorganisms and inactivates viruses.
This process can also kill bacterial spores but disinfectants used would require a long period of
time (e.g., 6 to 10 hours) to create sterilization. Such disinfectants are generally used on medical
devices but not on surfaces such as floors and benches. A much more similar process to this is
Intermediate-level disinfection except in the sense that the process is not capable of sterilization
even when disinfectants are applied for long period of time. This process is more commonly used in
the laboratory and for housekeeping purposes. Lastly, Low-level disinfection process inactivates
some viruses and kills most vegetative bacteria except M. tuberculosis. Disinfectants of this level
are generally used as hospital disinfectants or sanitizers.
Decontamination is the use of sterilization and disinfection techniques to free a person, object or
an area from potentially harmful microorganisms and render it safe for use (Block, 2001). In the
microbiology laboratory, decontamination aims to protect the laboratory worker, the environment
or anybody who had physical contact to the laboratory and its equipment (Chosewood & Wilson,
2009)..
2. There are types of solutions or reagents that cannot be heated but instead filtered to make them
devoid of contaminants. Give examples of these types of solutions.
Solutions that cant be autoclaved but can be filtered are mostly heat sensitive liquids such as
serum, antibiotic solutions, sugar solutions and urea solutions (Rao, 2008). Aside from this, special

culture media that are also prone to heat damage are also decontaminated through filtration which
can successfully isolate microbes but not mycoplasmas and viruses (Collins, Lyne, & Grange, 1995).

3. What are biosafety cabinets? How do they differ from laminar flow hoods? When are they used?
Biosafety cabinets (BSC) are entirely enclosed environments that are available in positive or
negative airflow (Elsevier Health Sciences, 2014). These are also known as glove boxes or isolator
hoods which differ from laminar flow hoods (LFH) in a sense that BSCs are applicable for both
infectious and non-infectious work whereas LFHs are only used for non-infectious work. In
particular, (University of Massachusetts Amherst, 2015) stated that BSCs are used when working
with potentially infectious and pathogenic microorganisms, as well as cell cultures and drug
formulations which should never be performed inside a laminar flow hood. While open face LFHs
only protect the product inside the hood, BSCs protect both the user and the items inside the
cabinet as it has a sash in front. Lastly, air in LFHs are blown towards the user while air in BSCs are
drawn away from the uses.

4. What are the modes of action of alcohol and Lysol? What other reagents maybe used to disinfect
surfaces?
According to Harisha (2006), Lysols active agent is orthophenylphenol, a phenolic compound
which has germicidal properties due to its ability to alter membrane permeability and protein
structure which can result to denaturation. Similar to Lysol, alcohols (either ethyl or isopropyl) also
cause denaturation of proteins as stated by Cappuccino & Sherman (2007). It also works as a lipid
solvent and a wetting agent (in tinctures) to increase wetting ability of other reagents Aside from
lysol and alcohol, other reagents that are used as disinfectants are chemicals like formaldehyde,
ethylene oxide, and halogens namely sodium hypochlorite and chloramine.

REFERENCES
[1] Chosewood, L. C & Wilson, D.E. (2009). Biosafety in Microbiological and Biomedical
Laboratories (5th ed). USA: Centers for Disease Control and Prevention, National Institutes of
Health
[2] Block, S.S. (2001). Disinfection, Sterilization and Preservation. Philadelphia: Lippincott Williams
and Wilkins
[3] Rao, S. (2008, June). Sterilization and Disinfection. Retrieved January 30, 2015, from
http://www.microrao.com/micronotes/sterilization.pdf
[4] Collins, C., Lyne, P. & Grange, J. (1995) Collins and Lyne's microbiological methods (7th ed.)
London: Butterworth-Heinemann.

[5a] Elsevier Health Sciences (2014). Mosby's Pharmacy Technician: Principles and Practice (4th
ed). Canada: Elsevier, Inc.
[5b] University of Massachusetts Amherst. (2015). Environmental Health & Safety Fact Sheet:
Biological Safety Cabinets vs. Laminar Flow Hoods. Retrieved January 30, 2015, from
https://www.ehs.umass.edu/sites/default/files/Fact-Sheet-BSC-vs-LFH.pdf
[6] Harisha, S. (2006). An Introduction to Practical Biotechnology (1st ed). New Delhi: Laxmi
Publications Ltd.
[7] Cappuccino, J., & Sherman, N. (2007). Microbiology: A Laboratory manual (8th ed). San
Francisco: Benjamin Cummings Publishing Company