Quantification of Escherichia coli in Purified Water

Using ATP Bioluminescence, Vibrio fischeri, and Heterotrophic Plate Count

Megan RosenbergerPhotos taken by student

Every year, more than 3.4 million people die as a result of water related diseases mainly caused by raw sewage contamination: Escherichia coli.

Africa Development Project

Carolina Biological

Chlorine Dioxide Iodine

Saris Activated Carbon

Vibrio fischeri ScaleAcceptable

Purify Water Source Again NOT Drinkable

Chlorine Dioxide

Purifies via oxidation and penetrates bacterial cell walls, killing the organism.

The purification takes four hours.

Potable Aqua

Iodine

Release the iodine bound in the tetraglycine hydroperiodide molecule

and penetrates the cell wall of the microorganism, rendering it inactive.

Potable Aqua

Saris

Can filter particles 0.2-1.0 μmNo research for these claims concerning E. coli

Africa Development Project

Activated Carbon

Has not been widely tested against E. coli

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Vibrio fischeri

Carolina Biological

Correlation of HPC with E. coli and ATP, Vibrio fischeri

PurposeThe purpose of my research is to determine which suggested and most economic water purification methods are most effective in removing TOP10 Escherichia coli from water, as well as to determine whether the quantification of the E. coli can be observed using an ATP bioluminescence, Vibrio fischeri.

Carolina Biological

Hypothesis

It is hypothesized that chlorine dioxide will perform the best because it has shown to be

effective against Cholera bacteria, which is similar to E. coli.

Research Institution

Center for Environmental Research and Education at Duquesne UniversityBiosafety Level 2 Lab

Photo taken by sponsor

Chlorine DioxideFor each 1mL of ultrapure water vial

prepared for chlorine dioxide, 2µL of the dissolved solution was added.

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IodineFor each 1mL of ultrapure water vial prepared for iodine, 1µL of the dissolved solution was added.

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Activated CarbonFifteen to twenty granules of activated carbon were added to each 1mL ultrapure water vial prepared.

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SarisUsing a syringe (without needle), 1mL was extracted from the vial and while holding one layer of the saris over the

syringe, it was released and spewed into the plate.

The quantification of the Vibrio fischeri was successful in detecting E. coli. All plates that were not inoculated with

TOP10 did emit light. However, all plates with the E. coli did not emit

light.

Photo taken by student

cfu/ml

Chlorine Dioxide

4.9

Iodine

1.5

Saris

Activated Carbon

1715.0

715.5Control

2675.9

Heterotrophic Plate Count Method Quantification

Chlorine Dioxide Iodine

Saris Activated Carbon

Iodine Chlorine Activated Carbon

Cloth0

0.05

0.1

0.15

0.2

0.25

0.3

0.35

0.4

0.45

0.5

Purification

Pric

e/un

it (c

ents

)

Conclusion

This study demonstrates that ATP bioluminescence is an effective and expedient method for detecting E. coli in contaminated

water. In correlation with HPC methods, it can be concluded that chlorine and iodine were both

significantly more effective in filtering E. coli than the carbon, cloth, and control.

Future Investigation

For future investigation, additional research should be made regarding the duration of

purification.

Carolina Biological

Works CitedSDI-Strategic Diagnostic, Inc. 2011. Luminescent Bacteria About Vibrio fischeri. Retrieved on 2011 From SDIX. Website: sdix.com. Hunter, Paul R.. 2003. Drinking Water and Diarrhoeal disease due to Escherichia coli. Journal of Water and Health. 01.2: 65-72. WHO-World Health Organization. 2011. Rainwater Harvesting. Retrieved on 9/11/11. From WHO. Website: who.int. Flinn Scientific, INC. Online Catalog. 2011. Bacterial Cultures, Vibrio fischeri. Retrieved on 9/12/11. From Flinn. Website flinnsci.com. Bartram, J., J. Cotruvo, M. Exner, C. Fricker, A. Glasmacher. Report. 2003. Heterotrophic Plate Counts and Drinking-Water Safety: The Significance of HPCs for Water Quality and Human Health.

AcknowledgementsDr. John Stolz, Director

Center for Environmental Research and Education at Duquesne University

 Lucas Eastham

Duquesne University Mrs. Catherine Greco

High School Sponsor Mr. Bob Wesolowski

Statistics Teacher My Parents