Molecular Tools: Molecular Tools: Engineering Engineering ApplicationsApplications

Environmental Environmental Engineering: State of the Engineering: State of the ArtArt ““Sanitary Engineering”Sanitary Engineering” ““Environmental Engineering”Environmental Engineering”

– Biological Processes:Biological Processes: WastewaterWastewater

– BOD removalBOD removal– Nutrient RemovalNutrient Removal

BioremediationBioremediation– Primary Substrate Degradation: ex: oil spillsPrimary Substrate Degradation: ex: oil spills– Co-metabolism: ex: chlorinated solventsCo-metabolism: ex: chlorinated solvents– Contaminant immobilization: ex: Heavy Contaminant immobilization: ex: Heavy

MetalsMetals

Other Engineering Other Engineering Fields:Fields: Biomedical Engineering: Biomedical Engineering:

– Targeted Drug DeliveryTargeted Drug Delivery– Gene TherapyGene Therapy

Chemical Engineering:Chemical Engineering:– Metabolic EngineeringMetabolic Engineering

Example: Heavy Metal Example: Heavy Metal RemediationRemediation

Tailings Dam

http://www.science.uwaterloo.ca/research/ggr/PermeableReactiveBarriers/PRBforAMD/PRBforAMD.html

Ecology of Sulfate-Ecology of Sulfate-Reducing CommunitiesReducing Communities

Polymers

Oligomers

Monomers

Organic Acids

Acetate

CO2

H2

Alcohols

CO2

SulfateReduction

Methanogenesis

CO2

SO42-

H2S

MeS(s)

Me2+

CH4 + CO2

EnzymaticHydrolysis

Fermentation

Hydrolytic processes Fermentative processes Methanogenic processes Sulfate-reduction processes

DGGE of PRBDGGE 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

PMa

DMa

DMb

WLa

WLb

#1a

#1b C5 C6

C17a

C17b

C34a

C34b

PMb

SSCP of PRBSSCP of PRB

B14 B12B29 B18B24B28 B25 B27

Columns 3,4

Real-Time PCRReal-Time PCR

Wetlands

Nor

mal

ized

DN

A C

once

ntra

tion

10-6

10-5

10-4

10-3

10-2

Initial Mixture

Column5

Column6

Column1,7

Column3,4

Desulfotomaculum Real-Time PCR: Eubacteria Normalized

Example: MTBE Example: MTBE BioremediationBioremediation

BTEXBTEXMTBEMTBE

Experimental Design:Experimental Design:

MTBE only

BTEX only

TBA W/

BTEX

TBAonly

MTBE W/

BTEX

Sterile control with

all 6 compounds

pH buffered sterilized nutrient solution

a well-characterized mixed culture

Concentration 50 mg/L

DGGE ProfilesDGGE ProfilesMTBE Only TBA only

Start Mid. End Start Mid. End

STD

Known MTBE degrader Hydrogenophaga

Known MTBE degrader “PM1-like”

Real Time PCR of Real Time PCR of MTBE-DegradersMTBE-Degraders

Eubacteria

Time, days

0 10 20 30 40

Co

nce

ntr

atio

n, (

ug

/uL

)

0

10

20

30

40MTBE onlyMTBE w/ BTEXTBA onlyTBA w/ BTEXBTEX only

"PM1 like"

Time, days

0 10 20 30 40

Co

nce

ntr

atio

n, (

ug

/uL

)

0.00

0.01

0.02

0.03

0.04MTBE onlyMTBE w/ BTEXTBA onlyTBA w/ BTEXBTEX only

Example: Fate of Example: Fate of Antibiotic GenesAntibiotic Genes >50% Antibiotics used in U.S. -> >50% Antibiotics used in U.S. ->

LivestockLivestock Antibiotics demonstrated to be in Antibiotics demonstrated to be in

Poudre River (Yang and Carlson)Poudre River (Yang and Carlson) Effect on Microbial Communities?Effect on Microbial Communities? Antibiotic Resistance Genes?Antibiotic Resistance Genes?

Other Areas:Other Areas:

Drinking Water PathogensDrinking Water Pathogens Activated Sludge UpsetActivated Sludge Upset Biological Phosphorus RemovalBiological Phosphorus Removal Assessment Tools in Assessment Tools in

BioremediationBioremediation

People, Names, Places:People, Names, Places:

Bruce Rittmann, Northwestern U., Bruce Rittmann, Northwestern U., Lutgarde Raskin, UI-UC, Nancy Love, Lutgarde Raskin, UI-UC, Nancy Love, Virginia Tech, Dan Oerther, U. Virginia Tech, Dan Oerther, U. Cincinnati, Dan Noguera, UNC, Francis Cincinnati, Dan Noguera, UNC, Francis de los Reyes, U Wisconsin- Madison, de los Reyes, U Wisconsin- Madison, Pedro Alvarez, U. IowaPedro Alvarez, U. Iowa

Carl Woese- UI-UC, Rudolf Aaman- Max Carl Woese- UI-UC, Rudolf Aaman- Max Planck Institute, Germany, Norm Pace- Planck Institute, Germany, Norm Pace- Boulder, David Stahl-, James Tiedje- Boulder, David Stahl-, James Tiedje- Mich. StateMich. State

Today’s Topics:Today’s Topics:

TCE-degrading CommunitiesTCE-degrading Communities Wastewater MicrobesWastewater Microbes Phosphate RemovalPhosphate Removal Food (beer) microbiologyFood (beer) microbiology

Flow CytometryFlow Cytometry