Industrial Microbiology

LAB Physical Agents of Control – Electromagnetic Radiation

Objectives
To study the effect of ultraviolet (UV) radiation on microorganism

Introduction
Certain forms of electromagnetic radiation are capable of producing a lethal effect on cells
and therefore can be used for microbial control. Electromagnetic radiations that possess
sufficient energy to be microbicidal are the short-wavelength radiation, that is, below
300nm. These include UV, gamma rays, and x-rays. The high wavelength radiations, those
above 300nm, have insufficient energy to destroy cells.

Materials, Culture, and Media

24 to 48 hours nutrient broth cultures of E. coli; inoculating loop, ultraviolet radiation
source, parafilm, 6 nutrient agar plates for each group.

Methodology
1. All plates were inoculated in a sterile manner, by means of streak inoculation.
2. Cover of each inoculated plate was labeled according to the exposure time to
ultraviolet radiation: 0 (control), 15 seconds, 30 seconds, 1 minute, 3 minutes, 5
minutes, and 5 minutes (irradiated covered control)
3. All inoculated plates were irradiated for the designated period of time by placed at
12 inches below the ultraviolet light sources. All petri dish covers was removed
except that of the 5-minutes irradiated control plate.
4. All plates were incubated in an inverted position for 4 to 5 days at 30°C.

Result
1. All nutrient agar plate cultures were observed for the amount of growth of microbe.
2. Observation was recorded as: 0 = no growth; 1+ = slight growth; 2+ = moderate
growth; 3+ = abundant growth.

Time of Irradiation
Microbial
Seconds Minutes
Species
0 15 30 1 3 5 5*
E. coli. 3+ 3+ 3+ 2+ 2+ 0 0
* irradiated, covered plate

Discussion
Control of microbial growth is accomplished through chemical, physical and
chemotherapeutic agents.

Physical agents of control include heat, freeze-drying, ultraviolet radiation and filtration.
Except for photosynthetic bacteria, most bacteria are harmed by ultraviolet radiation.
Although sunlight contains the complete spectrum of short to long wavelengths of light, it is
only the short invisible ultraviolet wavelengths that are injurious to these bacteria.
Ultraviolet radiation is strongly absorbed by proteins and nucleic acids in cells; hence, the
indications are that cellular damage involves changes in DNA. Depending on the dosage,
ultraviolet radiation may cause enzyme inactivation, genetic mutation or death of the cell.
The longer the exposure, the more microbes will kill.

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Examples: when microbes were treated with ultraviolet radiation, the radiation affects DNA
in their cell. (a) A normal DNA was converted into (b) an abnormal DNA molecule as the UV
light binds adjacent thymine molecules within the DNA to form (c) a thymine dimer. With its
thymine molecules bound in dimers, the DNA molecules cannot function properly and
cannot replicate. The microbe quickly dies.

Other kinds of radiation, includes gamma rays and X-rays which are highly penetrating to
cells and tissues and have potent antimicrobial effects (they are 10000 times more energetic
than UV light). After colliding with a target, ionizing radiation generates ions and other
reactive species from molecules including hydroxyl (free) OH- radicals. These free radicals
can cause irreversible breaks in DNA, proteins and enzymes.

Conclusion
Through this experiment, I am understood and aware of the effect of ultraviolet (UV)
radiation on microorganism.

References
Microbiology: A Laboratory Manual Seventh Edition (International Edition). Cappucino and
Sherman. Pearson. Benjamin Cummings

http://scienceprofonline.googlepages.com/LabExerciseandReport5.doc. (251008)

Microbes and Society: An Introduction to Microbiology. I. Edward Alcamo. Jones & Bartlett
Publishers, 2002. Boston, Massachusetts.

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0 second 15 seconds

30 seconds 1 minute

3 minutes 5 minutes

5 minutes*