Patel 1

Concentration and Duration of Ultraviolet Light Exposure on E. Coli

Aashka Patel aashka615@gmail.com
Marine Academy of Technology and Environmental Science, NJ
Introduction:
Nearly every 4-20 minutes bacteria reproduces (PNNL, 2010). Bacteria reproduce
asexually through a process called binary fission. Binary fission is when a single cell bacteria
divides into two identical daughter cells. The bacterial cell that grows longer, then splits into two
more daughter cells that each have the same DNA as the parent cell. Escherichia coli or E. coli
normally is found in an animal's intestinal tract. E. coli is able to sustain itself with and without
oxygen and comes in virulent and non-virulent strains. Virulence factors related to the
pathogenicity of ExPEC are numerous from those related to bacteria colonization to those related
to virulence (A. Alkeskas et al. , 2019). The virulence of E. coli is affected by many different
factors. Including adhesins, toxins, iron acquisition factors ( A. Alkeskas et. al, 2019). E. coli is
mostly found in contaminated food and water and produces a toxin that can damage the lining of
the small intestine.
Ultraviolet (UV) radiation is used in the medical field to kill bacteria. Ultraviolet
radiation is used for killing bacteria, phototherapy, suntanning, and a variety of other reasons
(Botzenhart et al., 2017). Different UV wavelengths and intensities are used for different
purposes. The different types of ultraviolet radiation are UVA, UVB, and UVC. UVA rays have
the longest wavelength, then UVB rays. UVC rays have the shortest wavelength. High doses of
UVA and UVB rays damages skin and can cause skin cancer. UVB is the cause of sunburns, it
harms the epidermis of your skin. UVB rays age skin overtime. UVB (290-320 nm) most
effectively produces skin cancer and sunburn (J. Jagger, 1983). UVA radiation causes skin
aging, like spots and wrinkles. UVC rays do not penetrate the Earth’s atmosphere naturally, but
there are still inorganic forms. UVC radiation can cause severe eye damage and severe burns on
the skin. UVB and UVC induced delayed apoptosis (Godar et al. , 2000). UVC rays are used to
kill bacteria. In this study, E. coli viability was measured to observe the effects of UV intensity
and exposure time. This study differs from other studies because the intensity and duration of
ultraviolet light are both being altered to see E. coli viability after being exposed to UVC light.
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Methodology:
1. Bacteria Quantity and Growth
a. 36 plates of E. coli are cultured using Coliscan and a sample of E. coli.
b. 0.5 mL of the E. coli sample is placed in the Coliscan bottle
c. Swirled the Coliscan bottle for 15 seconds, and then poured into the petri dish
(Figure 1).

Figure 1: 36 bottles and 36 petri

dishes were used to culture the E. coli.

d. Placed the petri dishes in the incubator for 48 hours at 37℃.

2. Bacteria Groups and Usage
a. The E. coli cultures were divided into 3 groups. Each group consisted of 12 plates
each.
b. Each group was subdivided into sections labeled : “A” or “B”. Each section had 6
plates.
c. Placed Group 1A under the ultraviolet lights for 5 minutes at 10 centimeters.
d. Placed Group 1B under the ultraviolet lights for 10 minutes at 10 centimeters.
e. Placed Group 2A under the ultraviolet lights for 5 minutes at 20 centimeters.
f. Placed Group 2B under the ultraviolet lights for 10 minutes at 20 centimeters.
g. Placed Group 3A under the ultraviolet lights for 5 minutes at 30 centimeters.
h. Placed Group 3B under the ultraviolet lights for 10 minutes at 30 centimeters.
3. Growth measurements
a. Recorded and analyzed data according to scale.
b. Created a scale 1-5. 1 being that the UV lights were the least effective, and 5
being the most effective (Figure 2).
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c. Each section per each group was scaled from 1-5 to determine the optimal use for
ultraviolet light on E. coli.

Figure 2: The scales used to measure how effective the UV

light was to the E. coli culture.
Results:
From the 36 E. coli cultures, 18 of the cultures visibly shrunk due to the ultraviolet light
(Table 2). According to the scale, Group 1B had the greatest change showing that all of the E.
coli bacteria was gone. The lowest amount of change happened in Group 3A. The graph shows
the groups and sections and how effective the ultraviolet was on the E. coli (Figure 3). In Group
2A and Group 2B both landed on 3 according to the scale because some of the plates showed
some change, but others showed no change.
Table 1: The values from the scale measured the viability of E. coli after the cultures were
placed under the UVC lights.
Distance (cm) Distance (cm)
Group 1 Group 2 Group 3 Group 1 Group 2 Group 3 (30
(10 cm) (20 cm) (30 cm) (10 cm) (20 cm) cm)
Time A A A Time B B B
5 minutes 4 3 1 10 minutes 5 3 2
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Figure 3: The efficiency of ultraviolet light in certain restraints. 5 being the most
efficient, and 1 being the least effective.
Table 2: Table used to describe if the E. coli bacteria shrunk
due to the ultraviolet light.

Group E. Coli shrink due to UV lights (yes/no)

1A yes
1B yes
2A yes
2B no
3A no
3B no
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Discussion:
The color change from purple to yellow in Group 1 occured because the UVC light was
placed the closest to the E. coli bacteria. Ultraviolet light is most effective when placed very
close to the E. coli. The E. coli in Group 3 had the least change because it was farthest from the
E. coli. The closer the E. coli culture was to the UVC light the more E. coli died. Eggs have
bacteria on the shells. To remove the bacteria. The eggs were placed under ultraviolet light
(Coufal et al., 2019). Similarly, the UVC lights were able to kill the E. coli bacteria. The UVC
lights killed the majority of the E. coli bacteria, when the E. coli was placed closely to the UVC
light.
Once the cultures were removed from the incubator after 48 hours, E. coli grew
everywhere and not in clusters or colonies. It was difficult to see and count how many colonies
were present in the culture. Instead of being able to see which colonies shrunk, the color changed
from purple to yellow signifying that the E. coli bacteria died. A scale was made instead to
determine the change in the cultures after placed under the ultraviolet light. It was also difficult
to turn off the UVC lights exactly at the specified time, which could have affected the results by
allowing discrepancies in UV exposure duration.
There was no noticable change in Group 3 because the E. coli culture was too far from
the ultraviolet light and it was not under the ultraviolet light for a long period of time. Ultraviolet
light is most effective when placed closest to the bacteria. Knowledge of UV radiation on
bacteria can contribute to optimize UV-based disinfection strategies (Environ. Sci. Technol.,
2013). UV radiation is used as a method to kill bacteria and disinfect surfaces. The closer the
ultraviolet light is to the bacteria the faster it will die.
Conclusion:
The E. coli culture that was placed under the ultraviolet light at 10 centimeters and spent
the most time under the ultraviolet light, which was for 20 minutes, had the least E. coli bacteria
left. It showed that the most effective way to use ultraviolet light is to keep it close and under the
bacteria for as long as possible to have the most effective way of getting rid of the bacteria. The
least effective way of decreasing the E. coli bacteria was keeping the ultraviolet light at a far
distance, and leaving it under for a short amount of time. The concentration and duration of UVC
light on E. coli bacteria is being tested to find the most successful method of sanitation of
bacteria.
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Acknowledgments:
I would like to acknowledge Dr. Wnek and MATES for providing me with the ability and
opportunity to conduct this research and for assistance throughout the process.
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