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Control of Microbial Growth

Surgery in the 19th century was not performed under aseptic conditions. Surgeons often operated wearing their street clothes, without washing their hands. Control of Microbial Growth means to inhibit or prevent growth of microorganisms.

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63 views

Control of Microbial Growth

Surgery in the 19th century was not performed under aseptic conditions. Surgeons often operated wearing their street clothes, without washing their hands. Control of Microbial Growth means to inhibit or prevent growth of microorganisms.

Uploaded by

Umar Kharisma
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Control of Microbial Growth (page 1)

(This chapter has 6 pages)


Kenneth Todar, PhD
Introduction
In the 19th century, surgery was risky and dangerous, and patients undergoing even the most routine operations were at
very high risk of infection. This was so because surgery was not performed under aseptic conditions. The operating room,
the surgeon's hands, and the surgical instruments were laden with microbes, which caused high levels of infection and
mortality.
Surgeons in the mid-1800s often operated wearing their street clothes, without washing their hands. They frequently used
ordinary sewing thread to suture wounds, and stuck the needles in the lapels of their frock coats in between patients.
Surgical dressings were often made up of surplus cotton or jute from the floors of cotton mills. It was against this
background that French scientist Louis Pasteur demonstrated that invisible microbes caused disease.

Louis Pasteur
Pasteur's work influenced the English surgeon Joseph Lister, who applied Pasteur's germ theory of disease to surgery, thus
founding modern antiseptic surgery. To disinfect, Lister used a solution of carbolic acid (phenol), which was sprayed around
the operating room by a handheld sprayer.

Joseph Lister

19th Century surgery using Lister's carbolic acid sprayer.


It was clear that Lister's techniques were effective in increasing the rates of surviving surgery, but his theories were
controversial because many 19th century surgeons were unwilling to accept something they could not see. Also, perhaps
another reason that surgeons were slow to pick up on Lister's methods was the fact that during surgery they were required
to breathe an irritating aerosol of phenol.
Control of Microbial Growth
The control of microbial growth is necessary in many practical situations, and significant advances in agriculture, medicine,
and food science have been made through study of this area of microbiology.
"Control of microbial growth", as used here, means to inhibit or prevent growth of microorganisms. This control is affected
in two basic ways: (1) by killing microorganisms or (2) by inhibiting the growth of microorganisms. Control of growth usually
involves the use of physical or chemical agents which either kill or prevent the growth of microorganisms. Agents which kill
cells are called cidal agents; agents which inhibit the growth of cells (without killing them) are referred to as static agents.
Thus, the term bactericidal refers to killing bacteria, and bacteriostatic refers to inhibiting the growth of bacterial cells. A
bactericide kills bacteria, a fungicide kills fungi, and so on.

In microbiology, sterilization refers to the complete destruction or elimination of all viable organisms in or on a substance
being sterilized. There are no degrees of sterilization: an object or substance is either sterile or not. Sterilization procedures
involve the use of heat, radiation or chemicals, or physical removal of cells.
Methods of Sterilization
Heat: most important and widely used. For sterilization one must consider the type of heat, and most importantly, the
time of application and temperature to ensure destruction of all microorganisms. Endospores of bacteria are considered
the most thermoduric of all cells so their destruction guarantees sterility.
Incineration: burns organisms and physically destroys them. Used for needles, inoculating wires, glassware, etc. and
objects not destroyed in the incineration process.
Boiling: 100o for 30 minutes. Kills everything except some endospores. To kill endospores, and therefore sterilize a
solution, very long (>6 hours) boiling, or intermittent boiling is required (See Table 1 below).
Autoclaving (steam under pressure or pressure cooker)
Autoclaving is the most effective and most efficient means of sterilization. All autoclaves operate on a time/temperature
relationship. These two variables are extremely important. Higher temperatures ensure more rapid killing. The usual
standard temperature/pressure employed is 121C/15 psi for 15 minutes. Longer times are needed for larger loads, large
volumes of liquid, and more dense materials. Autoclaving is ideal for sterilizing biohazardous waste, surgical dressings,
glassware, many types of microbiologic media, liquids, and many other things. However, certain items, such as plastics and
certain medical instruments (e.g. fiber-optic endoscopes), cannot withstand autoclaving and should be sterilized with
chemical or gas sterilants. When proper conditions and time are employed, no living organisms will survive a trip through
an autoclave.

Schematic diagram of a laboratory autoclave in use to sterilize microbiological culture medium. Sterilization
of microbiological culture media is is often carried out with the autoclave. When microbiological media are
prepared, they must be sterilized and rendered free of microbial contamination from air, glassware, hands,
etc. The sterilization process is a 100% kill, and guarantees that the medium will stay sterile unless exposed
to contaminants.

An autoclave for use in a laboratory or hospital setting.


Why is an autoclave such an effective sterilizer? The autoclave is a large pressure cooker; it operates by using steam under
pressure as the sterilizing agent. High pressures enable steam to reach high temperatures, thus increasing its heat content
and killing power. Most of the heating power of steam comes from its latent heat of vaporization. This is the amount of heat
required to convert boiling water to steam. This amount of heat is large compared to that required to make water hot. For
example, it takes 80 calories to make 1 liter of water boil, but 540 calories to convert that boiling water to steam.
Therefore, steam at 100 C has almost seven times more heat than boiling water.
Moist heat is thought to kill microorganisms by causing denaturation of essential proteins. Death rate is directly
proportional to the concentration of microorganisms at any given time. The time required to kill a known population of
microorganisms in a specific suspension at a particular temperature is referred to as thermal death time (TDT).
Increasing the temperature decreases TDT, and lowering the temperature increases TDT. Processes conducted at high
temperatures for short periods of time are preferred over lower temperatures for longer times.
Environmental conditions also influence TDT. Increased heat causes increased toxicity of metabolic products and toxins.
TDT decreases with pronounced acidic or basic pHs. However, fats and oils slow heat penetration and increase TDT. It must
be remembered that thermal death times are not precise values; they measure the effectiveness and rapidity of a
sterilization process. Autoclaving 121C/15 psi for 15 minutes exceeds the thermal death time for most organisms except
some extraordinary sporeformers.
Dry heat (hot air oven): basically the cooking oven. The rules of relating time and temperature apply, but dry heat is not
as effective as moist heat (i.e., higher temperatures are needed for longer periods of time). For example 160 o/2hours or
170o/1hour is necessary for sterilization. The dry heat oven is used for glassware, metal, and objects that won't melt.

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