Molecular Tools: Engineering Applications

Environmental Engineering: State of the Art “Sanitary Engineering” “Sanitary Engineering” “Environmental Engineering” “Environmental Engineering” –Biological Processes: Wastewater Wastewater –BOD removal –Nutrient Removal Bioremediation Bioremediation –Primary Substrate Degradation: ex: oil spills –Co-metabolism: ex: chlorinated solvents –Contaminant immobilization: ex: Heavy Metals

Other Engineering Fields: Biomedical Engineering: Biomedical Engineering: –Targeted Drug Delivery –Gene Therapy Chemical Engineering: Chemical Engineering: –Metabolic Engineering

Example: Heavy Metal Remediation Tailings Dam

Ecology of Sulfate- Reducing Communities Polymers Oligomers Monomers Organic Acids Acetate CO 2 H2H2 Alcohols CO 2 Sulfate Reduction Methanogenesis CO 2 SO 4 2- H2SH2S MeS (s) Me 2+ CH 4 + CO 2 Enzymatic Hydrolysis Fermentation Hydrolytic processes Fermentative processes Methanogenic processes Sulfate-reduction processes

DGGE of PRB PM - standard DM - inocula from dairy manure WL - inocula from wetland #1 - initial mixture C5 - column 5 C6 - column 6 C17 - mixture of column 1 and 7 C34 - mixture of column 3 and 4 PM a DM a DM b WL a WL b #1 a #1 b C5C6 C17 a C17 b C34 a C34 b PM b

SSCP of PRB B14B12B29 B18 B24 B28B25 B27 Columns 3,4

Real-Time PCR

Example: MTBE Bioremediation BTEX MTBE

Experimental Design: MTBE only BTEX only TBA W/ BTEX TBA only MTBE W/ BTEX Sterile control with all 6 compounds pH buffered sterilized nutrient solution a well-characterized mixed culture Concentration 50 mg/L

DGGE Profiles MTBE Only TBA only Start Mid.End Start Mid. End STDSTD Known MTBE degrader Hydrogenophaga Known MTBE degrader “PM1-like”

Real Time PCR of MTBE- Degraders

Example: Fate of Antibiotic Genes >50% Antibiotics used in U.S. -> Livestock >50% Antibiotics used in U.S. -> Livestock Antibiotics demonstrated to be in Poudre River (Yang and Carlson) Antibiotics demonstrated to be in Poudre River (Yang and Carlson) Effect on Microbial Communities? Effect on Microbial Communities? Antibiotic Resistance Genes? Antibiotic Resistance Genes?

Other Areas: Drinking Water Pathogens Drinking Water Pathogens Activated Sludge Upset Activated Sludge Upset Biological Phosphorus Removal Biological Phosphorus Removal Assessment Tools in Bioremediation Assessment Tools in Bioremediation

People, Names, Places: Bruce Rittmann, Northwestern U., Lutgarde Raskin, UI-UC, Nancy Love, Virginia Tech, Dan Oerther, U. Cincinnati, Dan Noguera, UNC, Francis de los Reyes, U Wisconsin- Madison, Pedro Alvarez, U. Iowa Bruce Rittmann, Northwestern U., Lutgarde Raskin, UI-UC, Nancy Love, Virginia Tech, Dan Oerther, U. Cincinnati, Dan Noguera, UNC, Francis de los Reyes, U Wisconsin- Madison, Pedro Alvarez, U. Iowa Carl Woese- UI-UC, Rudolf Aaman- Max Planck Institute, Germany, Norm Pace- Boulder, David Stahl-, James Tiedje- Mich. State Carl Woese- UI-UC, Rudolf Aaman- Max Planck Institute, Germany, Norm Pace- Boulder, David Stahl-, James Tiedje- Mich. State

Today’s Topics: TCE-degrading Communities TCE-degrading Communities Wastewater Microbes Wastewater Microbes Phosphate Removal Phosphate Removal Food (beer) microbiology Food (beer) microbiology

Flow Cytometry