Title page for ETD etd-01072002-093548

Type of Document Master's Thesis
Author Wimmer, Robert Francis
Author's Email Address rwimmer@vt.edu
URN etd-01072002-093548
Title Development of a Biosensor to Predict Activated Sludge Deflocculation, and the Link Between Chlorination and Potassium Efflux
Degree Master of Engineering
Department Environmental Engineering
Advisory Committee
Advisor Name Title
Love, Nancy G. Committee Chair
Love, Brian J. Committee Member
Novak, John T. Committee Member
  • biosensor
  • potassium efflux
  • chlorine
  • microfluidic
  • activated sludge
  • glutathione
Date of Defense 2001-12-21
Availability unrestricted
In an effort to provide wastewater treatment operators with the capability to be proactive in assessing and solving deflocculation events, this study has tested the components of a biosensor to predict deflocculation and investigated the mechanistic cause of deflocculation relating to chlorination of activated sludge cultures. In order to effectively manage upset events, it is necessary to know the source of an upset and the causative mechanism that the source initiates.

The Glutathione-gated potassium efflux (GGKE)induced activated sludge deflocculation biosensor incorporates novel microtechnology with a whole cell biological element to predict deflocculation from electrophilic sources. This sensor utilizes microfluidic channels to conduct influent wastewater across a biofilm of Eschericia coli K 12 and monitors the bacterial response to the influent. The bacterial response, which is efflux of K+ ion from the cytoplasm, is monitored with a fluorescence-based sensor called an optode. The components of the system satisfy the project requirements, which include minimal expense (both operation and manufacture), on-line capability and minimal maintenance. The research conducted to date demonstrates the ability of the components of the biosensor to fulfill the design requirements. The optode K+ detector successfully measured an increase in soluble K+ following the exposure of E. coli K-12 to the electrophile N ethyl malemide. The manufacture of the microfluidic device has been completed and the device has demonstrated the ability to conduct influent under negative pressure across an established biofilm with the optode in place. The establishment of a biofilm under expected hydrodynamic conditions has also been completed. Future research efforts will include integrating the components of the biosensor into a working prototype that will be capable monitoring the reaction of bacteria to the presence of electrophilic compounds in wastewater. Sensors of this nature will provide operators with the early warning necessary to be proactive against toxic upsets rather than reactive.

The knowledge needed to create a biosensor resides in the identification of bacterial response mechanisms that cause upset events in wastewater treatment facilities. The biosensor that has been developed relies on the discovery of the link between electrophile-induced GGKE and activated sludge deflocculation. Research has been concluded, which expands the role of GGKE and activated sludge deflocculation to include chlorine-induced GGKE. Through a series of laboratory-scale reactors, a relationship has been established between chlorine addition to control filamentous bulking, increased soluble K+ levels and an increase in effluent suspended solids . The results demonstrate that the addition of chlorine to control filamentous bulking may elicit the GGKE mechanism, initiating activated sludge deflocculation, similar to observations of chlorination at full-scale activated sludge wastewater treatment facilities. Establishing a mechanistic cause of deflocculation related to chlorination will permit operators to apply chlorine in a manner that may avoid deflocculation, rather than reacting to deflocculation after it has occurred.

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