Topic 1- Control of Microbial Growth (Introduction
Considerations in Microbial Control
Controlling microbial growth is important in many
fields but the degree of acceptable microbial levels
can be quite different. Ever since microbes were
shown to cause diseases, people have invented
different techniques to control their spread.
Controlling microbial growth is important in the
medical field, pharmaceutical and biotechnology
industries, academic research, and food industry.
Each antimicrobial substance or agent achieves a
different level of microbial elimination by a certain
mechanism.
TYPES OF MICROBIAL CONTROL
Sterilization (or sterilisation ) is a term referring to
any process that eliminates (removes) or kills all
forms of microbial life, including transmissible
agents (such as fungi, bacteria, viruses, and spore
forms) present on a surface, contained in a fluid, in
medication, or in a compound. Sterilization can be
achieved by applying the proper combinations of
heat, chemicals, irradiation,
high pressure, and filtration.
Chemical agents that can eliminate or suppress
microbial life are separated in different groups
based on their use.
Disinfectants are substances that are applied to
non-living objects to destroy microorganisms that
are living on them. Disinfection does not
necessarily kill all microorganisms, especially
resistant bacterial spores, so it is less effective than
sterilisation. Disinfectants are different from other
antimicrobial agents such as antibiotics, which
destroy microorganisms within the body.
Disinfectants are also different from biocides, as
these are intended to destroy all forms of life, not
just microorganisms. Disinfectants work by
destroying the cell wall of microbes or
interfering with their metabolism
Antiseptics are antimicrobial substances that are
applied to living tissue or skin to reduce the
possibility of infection, sepsis, or putrefaction.
Antiseptics are generally distinguished from
antibiotics by the latter’s ability to be transported
through the lymphatic system to destroy bacteria
within the body, and from disinfectants, which
destroy microorganisms found on non living
objects.
The term antibiotic was first used in 1942 by
Selman Waksman and his collaborators in journal
articles to describe any substance produced by a
microorganism that is antagonistic to the growth of
other microorganisms in high dilution. This
definition excluded substances that kill bacteria, but
are not produced by microorganisms (such as
gastric juices and hydrogen peroxide). It also
excluded synthetic antibacterial compounds such
as the sulfonamides. With advances in medicinal
chemistry, most of today’s anti-bacterials chemically
are semisynthetic modifications of various natural
compounds. Many antibacterial compounds are
classified on the basis of chemical or biosynthetic
origin into natural, semisynthetic, and synthetic.
Another classification system is based on biological
activity. In this classification, antibacterials are
divided into two broad groups according to their
biological effect on microorganisms: bactericidal
agents kill bacteria; and bacteriostatic agents slow
down or stall bacterial growth. Microbicides which
destroy virus particles are called viricides or
antivirals.
LEVEL OF MICROBIAL PRESENCE
Joseph Lister: Joseph Lister was one of the first to
use aseptic techniques during surgeries.
The degree of acceptable microbial presence can
differ based on the circumstances. Sterilization as a
definition means that all life was terminated,
whereas sanitization and disinfection terminates
selectively and partially. Both sanitization and
disinfection reduce the number of targeted
pathogenic organisms to what are considered
―acceptable‖ levels – levels that a reasonably
healthy, intact body can deal with. In general,
surgical instruments and medications that enter an
already aseptic part of the body (such as the
bloodstream, or penetrate the skin) must be
sterilized to a high sterility assurance level (SAL).
Examples of such instruments include scalpels,
hypodermic needles, and artificial pacemakers. For
example, medical device manufacturers design
their sterilization processes for an extremely low
SAL. Their ―one in a million‖ devices should be
non-sterile. This is also essential in the
manufacture of parenteral pharmaceuticals.
Preparation of injectable medications and
intravenous solutions for fluid replacement therapy
requires not only a high sterility assurance level,
but also well-designed containers to prevent entry
of adventitious agents after the initial product
sterilization.
Food preservation is another field where the
presence of microorganisms is taken under
consideration. The process usually involves
preventing the growth of bacteria, fungi (such as
yeasts), and other microorganisms (although some
methods work by introducing benign bacteria or
fungi to the food).
Rate of Microbial Death
The rate of microbial death is used to develop
standard protocols for sterilization in many
industries. The rate of microbial death can be
determined. It is important in order to develop
standard protocols for disinfection which will
facilitate the sterilization routine in many industries.
The goal is to find out what is the minimum time
needed to achieve an acceptable level of
sterilization for a specific purpose. The killing agent
can be different (e.g., heat, chemical with certain
concentration) depending on the specific
application.
When the killing factor is heat, the phrase thermal
death can be used. Thermal death time is a
concept used to determine how long it takes to kill
specific bacteria at a specific temperature. It
was originally developed for food canning and has
found applications in cosmetics, and in producing
salmonella-free feeds for animals (e.g. poultry, and
pharmaceuticals).
Relative Resistance of Microbes
Different microbial structures and types of microbial
cells have different level of resistance to
antimicrobial agents.
Different microbial structures and types of microbial
cells have different level of resistance to
antimicrobial agents used to eliminate them.
Endospores are considered the most resistant
structure of microbes. They are resistant to most
agents that would normally kill the vegetative cells
from which they formed. Nearly all household
cleaning products, alcohols, quaternary ammonium
compounds and detergents have little effect.
However, alkylating agents (e.g. ethylene oxide),
and 10% bleach are effective against endospores.
Endospores are able to survive boiling at 100°C for
hours. Prolonged exposure to ionizing radiation,
such as x-rays and gamma rays, will also kill most
endospores.
Certain bacterial species are more resistant to
treatment than others. Mycobacterial infections are
notoriously difficult to treat. The organisms are
hardy due to their cell wall, which is neither truly
Gram negative nor positive. In addition, they are
naturally resistant to a number of antibiotics that
disrupt cell-wall biosynthesis, such as penicillin.
Due to their unique cell wall, they can survive long
exposure to acids, alkalis, detergents, oxidative
bursts, lysis by complement, and many antibiotics.
Most mycobacteria are susceptible to the antibiotics
clarithromycin and rifamycin, but antibiotic-resistant
strains have emerged.
Protozoa cysts are quite hard to eliminate too. As
cysts, protozoa can survive harsh conditions, such
as exposure to extreme temperatures or harmful
chemicals, or long periods without access to
nutrients, water, or oxygen for a period of time.
Being a cyst enables parasitic species to survive
outside of a host, and allows their transmission
from one host to another. Protozoa cells are also
hardy to eliminate. Gram-negative bacteria have
high natural resistance to some antibiotics.
Examples include Pseudomonas spp. which are
naturally resistant to penicillin and the majority of
related beta-lactam antibiotics. This ability to thrive
in harsh conditions is a result of their hardy cell
wall that contains porins. Their resistance to most
antibiotics is attributed to efflux pumps, which pump
out some antibiotics before the antibiotics are able
to act. Staphylococcus aureus is one of the major
resistant pathogens. Found on the mucous
membranes and the human skin of around a third
of the population, it is extremely adaptable to
antibiotic pressure. It was one of the earlier bacteria
in which penicillin resistance was found—in 1947,
just four years after the drug started being
mass-produced. Methicillin-resistant
Staphylococcus aureus (MRSA) was first detected
in Britain in 1961, and is now ―quite common‖ in
hospitals. A recent study demonstrated that the
extent of horizontal gene transfer among
Staphylococcus is much greater than previously
expected—and encompasses genes with functions
beyond antibiotic resistance and virulence, and
beyond genes residing within the mobile genetic
elements. Fungal cells as well as spores are more
susceptible to treatments. Vegetative bacterial and
yeast cells are some of the easiest to eliminate with
numerous agents and methods. Viruses, especially
enveloped ones, are relatively easy to treat
successfully with chemicals due to the presence of
lipids