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Laboratory Exercise No.

STERILIZATION PROCEDURES

RESULTS

In this experiment, four sterilization techniques namely the moist heat, dry heat, incineration,
and autoclave were introduced. This experiment focused more on the autoclave technique for
sterilization. The different parts of an autoclave are shown in Figure 1.

Figure 1. The basic parts of an autoclave.

DISCUSSIONS

Sterilization plays an essential role in the complete destruction or removal of all microorganisms
that could contaminate materials and thus constitute to a health hazard. Microorganisms vary in their
resistance to destruction by physical or chemical means. Normally, sterilization is best achieved by
physical methods such as moist or dry heat, which requires less time and reliable.

Moist heat in the form of pressurized steam is regarded as the most dependable method for the
destruction of all forms of life, including bacterial spores. This method is incorporated into a device
called the autoclave (Hussain, 1970).

Autoclaving is one of the techniques used in moist heat sterilization. The equipment used is
called an autoclave. An autoclave sterilizes using pressurized steam heat. The principle is to subject the
bacteria to a very high temperature of steam, which can be heated to a much greater temperature than
the normal boiling point of water under pressure, causing the coagulation of cell proteins of the
microorganism rapidly and efficiently. When heated, wet proteins release free -SH groups and form small
peptide chains. These chains are motile and able to establish new bonds between themselves and form
new complex structure different from the original proteins. The higher the pressure created inside the
autoclave, the higher would be the attainable temperature inside it.

Autoclaving is mostly done under 15 lbs per sq. inch pressure at 121 oC for a period of 15-20
mins. At the point of condensation steam liberates thermal energy equal to its heat of vaporization. As
the temperature increases, steam gets energized increasing its penetration capability (Pingali, 2011).

Autoclaving in this experiment was used to sterilize nutrient agar media. It can also be used to
sterilize instruments, dressings, intravenous equipment, applicators, solutions, syringes, transfusion
equipment, and numerous other items that can withstand high temperatures and pressures. The
autoclave also has certain limitations. For example, some plastic ware melts in the high heat, and sharp
instruments often become dull.

Dry heat was the first method of sterilization, and is a longer process than moist heat
sterilization. The destruction of microorganisms through the use of dry heat is a gradual phenomenon.
With longer exposure to lethal temperatures, the number of killed microorganisms increases. In dry
heating, the bacteria are burned to death or oxidized. Dry, hot air is much less effective in transferring
heat than moist heat. This is why microorganisms are much more able to withstand heat in a dry state
(Poonam, 2016).

Dry-heating with an oven works similarly like the autoclave, but without water and pressure.
Basically, the bacteria are subjected to lethal temperatures while also desiccating them. However, this
process takes longer than autoclaving (Emerah, 2017). Sterilization using dry heat is usually done at 160-
180C. Sterilization by dry-heat kills microorganisms by oxidizing their proteins and other chemical
constituents. When dry proteins are heated, the polar groups in their peptide chains are less active due
to absence of water and their motility is also much reduced (Garg, n.d.).

Another technique is incineration. It is a waste treatment process that involves the combustion
of organic substances contained in waste materials. This method burns any organism to ash. It is used to
sterilize medical and other bio hazardous waste before it is discarded with non-hazardous waste.
Bacteria incinerators are mini furnaces used to incinerate and kill off any micro organisms that may be on
an inoculating loop or wire (Poonam, 2016).

Tyndalization, a dry heat technique, it is an outdated and lengthy process designed to reduce the
level of activity of sporulating bacteria that are left by a simple boiling water method. The process
involves boiling for a period at atmospheric pressure, cooling, incubate for a day, then repeating the
process a total of three to four times. The incubation periods are to allow heat-resistant spores surviving
the previous boiling period to germinate to form the heat-sensitive vegetative which can be killed by the
next boiling step. This is effective because many spores are stimulated to grow by the heat shock. The
procedure only works for media that can support bacterial growth, and will not sterilize non-nutritive
substrates like water (Poonam, 2016).

Dry heat sterilization is applicable to materials that are damaged by high pressure or moisture
and can withstand high temperatures. Dry heat does not cause corrosion of metal instruments; thus it is
well suited for the sterilization of metal instruments but cannot be used for sterilization of fabrics,
plastics or rubber-ware. It is also suitable for glassware in the lab. The final category is materials that
steam cannot penetrate or must remain dry: This includes pharmaceutical powders that need to remain
dry, oils and substances that aren't penetrated by steam (Finkiel, 2017).

Limitations of dry heat includes very high temperature requirement, higher power consumption
and cost compared to autoclave, therefore much higher running costs, and it can damage some
instruments.
In sterilization, the higher the temperature the less amount of time is required and vice versa.
Steam or moisture is able to transfer heat far more rapidly than dry heat alone. Under pressure,
moisture will increase the temperature quickly. This high temperature will cause protein denaturation. It
is also more effective than dry heat for killing microorganisms in a sense that it is able to penetrate
microbial cells more efficiently. In addition, moist heat kills cells more rapidly and at lower temperatures.
Surgical instruments are ideally sterilized using moist heat while glassware, metal instruments, paper
wrapped things and syringes are effectively sterilized through dry heat (Cheprasov, n.d.).

In conclusion, the higher the temperature the less amount of time is required for sterilization.
Autoclaving is the most reliable method for the destruction of all forms of life, including bacterial spores.
It coagulates the cell proteins of the microorganism rapidly and efficiently. However, it has certain
limitations like in plastic ware that melts in high heat, and sharp instruments often become dull. In dry
heat technique, takes a longer process than that of the moist heat sterilization. It is applicable to
materials that are damaged by high pressure or moisture and can withstand high temperatures. It is well
suited for the sterilization of metal instruments but cannot be used for sterilization of fabrics, plastics or
rubber-ware.

REFERENCES

Hussain, M. (1970, January 01). Autoclaving: Real Sterilization. Retrieved September 10, 2017, from
http://microbiologyon-line.blogspot.com/2009/08/autoclaving-real-sterilization.html

Ex 10: Moist and Dry Heat. (n.d.). Retrieved September 12, 2017, from https://quizlet.com/73916836/ex-
10-moist-and-dry-heat-flash-cards/

Poonam. (2016, April 20). Discussion about sterilization. Retrieved September 10, 2017, from
http://www.biyanicolleges.org/discussion-about-sterilization/

Finkiel, M., Ph.D. (2017, January 17). Sterilization by Heat. Retrieved September 12, 2017, from
https://tuttnauer.com/blog/sterilization-heat

(n.d.). Retrieved September 12, 2017, from http://study.com/academy/lesson/autoclaves-and-moist-


heat-sterilization-use-with-surgical-tools.html

The Sterilization Process (Autoclaves). (n.d.). Retrieved September 12, 2017, from
http://www.eurotherm.com/sterilization

(n.d.). Retrieved September 12, 2017, from


http://www.biologydiscussion.com/bacteria/sterilization/moist-heat-sterilization-and-dry-heat-
sterilization/55169

Prasad, B. V., & Pingali, S. (2016). What is the principle involved in autoclave ? Retrieved September 12,
2017, from http://www.pharmainfo.net/answers/microbiology/what-principle-involved-autoclave

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