Occurrence, Distribution and Rate of Attachment of E. coli to Natural and Engineered Porous Surfaces

Mentor 1

David Garman

Mentor 2

Marcia Silva

Location

Union Wisconsin Room

Start Date

29-4-2016 1:30 PM

End Date

29-4-2016 3:30 PM

Description

We are developing a novel method for evaluating biofilm growth on the surface of natural and engineered porous particles using environmental Escherichia coli (E. coli) as a model organism. E. coli was grown on modified mTEC agar from return activated sludge samples collected at Milwaukee Metropolitan Sewerage District. Colonies were then picked and inoculated in Luria-Bertani Broth and incubated to form a liquid culture. E. coli cultures were then diluted and introduced to natural and engineered porous particles. Images were taken using a Zeiss LSM 710 Confocal microscope and beginning fluorescence intensity values were observed at 0 hours. The porous particles and E. coli mixture were then incubated at 37°C and images and fluorescence intensity values were obtained 24 hours later and again one week later. By measuring the fluorescence intensity values of a serial dilution of E. coli, we were able to build a calibration curve between concentrations of bacteria using the standard EPA Method 1603 versus the fluorescence intensity. Over the course of a week, the fluorescence intensity values increased from 8 to 250 in the 1:10 dilution, which we correlated to an increase in colony forming units per milliliter present in the growing biofilm that had adhered to the natural porous particle surface. These results indicate that natural porous particles sustained biofilm growth. The same procedure will be performed with engineered porous particles. The understanding of biofilm architecture and growth on the surface of porous materials and new fluorescence microscopy techniques will aide in the development of logical surface design of porous materials with high antibacterial potential.

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Apr 29th, 1:30 PM Apr 29th, 3:30 PM

Occurrence, Distribution and Rate of Attachment of E. coli to Natural and Engineered Porous Surfaces

Union Wisconsin Room

We are developing a novel method for evaluating biofilm growth on the surface of natural and engineered porous particles using environmental Escherichia coli (E. coli) as a model organism. E. coli was grown on modified mTEC agar from return activated sludge samples collected at Milwaukee Metropolitan Sewerage District. Colonies were then picked and inoculated in Luria-Bertani Broth and incubated to form a liquid culture. E. coli cultures were then diluted and introduced to natural and engineered porous particles. Images were taken using a Zeiss LSM 710 Confocal microscope and beginning fluorescence intensity values were observed at 0 hours. The porous particles and E. coli mixture were then incubated at 37°C and images and fluorescence intensity values were obtained 24 hours later and again one week later. By measuring the fluorescence intensity values of a serial dilution of E. coli, we were able to build a calibration curve between concentrations of bacteria using the standard EPA Method 1603 versus the fluorescence intensity. Over the course of a week, the fluorescence intensity values increased from 8 to 250 in the 1:10 dilution, which we correlated to an increase in colony forming units per milliliter present in the growing biofilm that had adhered to the natural porous particle surface. These results indicate that natural porous particles sustained biofilm growth. The same procedure will be performed with engineered porous particles. The understanding of biofilm architecture and growth on the surface of porous materials and new fluorescence microscopy techniques will aide in the development of logical surface design of porous materials with high antibacterial potential.