An Overview of Bacterial Source Tracking - Methods and

Applications

Douglas Moyer & Kenneth HyerU.S. Geological Survey

Richmond, Virginia

October 17, 2003

An Overview of Bacterial Source Tracking - Methods and

Applications

Douglas Moyer & Kenneth HyerU.S. Geological Survey

Richmond, Virginia

October 17, 2003

303(D) List of Impaired Waters

Determine the sources ofDetermine the sources offecal coliform bacteriafecal coliform bacteriacausing the violationscausing the violations

• More than 600 stream/riversMore than 600 stream/rivers listed as impairedlisted as impaired

• More than 50% listed for More than 50% listed for violations of the fecal violations of the fecal coliform bacteria standardcoliform bacteria standard

Bacterial Source TrackingBacterial Source Tracking

WaterSamples

SourceSamples

IndicatorOrganism

IndicatorOrganism

patternC

patternB

patternA

patternDog

patternHuman

patternGoose

UNKNOWN KNOWN

BST Methods Comparison Study

• Study involves a direct comparison of 7 BST methods. Each researcher is provided a collection of known library isolates. Evaluation is based on the utility of each method to identify 200

blind isolates (which were from known sources).

• Five Genotypic Methods (and investigators)– Ribotyping using two different enzyme sets

(George Lukasik, Mansour Samadpour)– Pulsed-field Gel Electrophoresis

(West Virginia Department of Agriculture)– rep-PCR using two different primer sets

(Howard Kator, Don Stoeckel)

• Two Phenotypic Methods (and investigators)– Antibiotic Resistance Analysis (Bruce Wiggins)– Carbon Substrate Utilization (Chuck Hagedorn)

Methods Comparison - Conclusions

• In a general sense, we found that:-In this study, under these conditions… These results may not be universal!-All methods did not produce comparable results; some methods were more

successful than others.-Results will be published this winter, likely in Environmental Science and

Technology. A draft manuscript is currently in the review process.• This is one of a few comparison studies being done. Need to see

what some of these other comparison studies determine...• Without stepping through the results, we can offer these

conclusions/recommendations:-Perform considerable QA/QC in your BST work! This may include: (1) analyzing blind collections of known isolates,

(2) use of multiple BST methods, and (3) the use of other tracers to support the BST work.

-Source tracking has tremendous potential, just be cautious in your application of this new technology.

Field Application of BST

• Three Streams included on the DEQ 303(d) list:Accotink Creek, Christians Creek, and Blacks Run

• Field Data Collection– Water-sample collection

• Baseflow - 8 samples every 6 weeks • Stormflow - 10 samples during 5 events

– Source Samples

• Bacteria Source Tracking Analysis – Ribotyping

• TMDL Development Using HSPF

BST Results:BST Results:By Individual ContributorBy Individual Contributor

Blacks RunBlacks Run

Christians CreekChristians Creek

Accotink CreekAccotink Creek

00

55

1010

1515

2020

2525

3030

3535G

oo

seG

oo

se

Hu

ma

nH

um

an

Do

gD

og

Du

ckD

uck

Ca

tC

at

Se

a G

ull

Se

a G

ull

Ra

cco

on

Ra

cco

on

Ca

ttle

Ca

ttle

De

er

De

er

Ho

rse

Ho

rse

Po

ultr

yP

oul

try

Pe

rce

nt o

f Kn

ow

nP

erc

en

t of K

no

wn

Validation of BST:Validation of BST:Human SignatureHuman Signature

Accotink CreekAccotink Creek Christians CreekChristians Creek Blacks RunBlacks Run

CaffeineCaffeine

00

0.050.05

0.10.1

0.150.15

0.20.2

0.250.25

Ca

ffein

e (

µg

/L)

Ca

ffein

e (

µg

/L)

CotinineCotinine

00

0.010.01

0.020.02

0.030.03

0.040.04

Co

tinin

e (

µg

/L)

Co

tinin

e (

µg

/L)

OtherOther21.1%21.1%

OtherOther12.0%12.0%

Accotink Creek, BST ResultsAccotink Creek, BST Results

(N=279)(N=279)

DogDog13.3%13.3%

DogDog9.0%9.0%

DeerDeer1.4%1.4%

DeerDeer10.0%10.0%

Four Mile Run, BST ResultsFour Mile Run, BST Results

(N=278)(N=278)

WaterfowlWaterfowl38.7%38.7%

WaterfowlWaterfowl37.0%37.0%

RaccoonRaccoon5.4%5.4%

RaccoonRaccoon15.0%15.0%

Results of BST:Results of BST:Comparison of Accotink Creek and Four Mile RunComparison of Accotink Creek and Four Mile Run

HumanHuman20.1%20.1%

HumanHuman17.0%17.0%

Fecal Coliform Modeling Process

ShallowShallowSub-SurfaceSub-Surface

StorageStorage

DeeperDeeperSub-SurfaceSub-Surface

StorageStorage

Wash-offWash-off by Overlandby Overland

FlowFlow

Release of Release of Bacteria withBacteria with

InterflowInterflow

PointPointSourceSource

Fecal Coliform/ManureFecal Coliform/ManureApplication/DepositionApplication/Deposition

LandLandSurfaceSurface

Die

-off

Die

-off

Release of Release of Bacteria withBacteria with

BaseflowBaseflow

Blacks R

un

Fecal Coliform ConcentrationFecal Coliform Concentration& BST Data & BST Data

Fecal Coliform Calibration

00

55

1010

1515

2020

2525

3030

3535

4040G

OO

SE

GO

OS

E

HU

MA

NH

UM

AN

DO

GD

OG

DU

CK

DU

CK

CA

TC

AT

RA

CC

OO

NR

AC

CO

ON

DE

ER

DE

ER

MU

SK

RA

TM

US

KR

AT

Observed BSTObserved BST

Initial Model CalibrationInitial Model Calibration

Final Model CalibrationFinal Model Calibration

Per

cent

Sig

natu

reP

erce

nt S

igna

ture

USGS Contact InformationUSGS Contact Information

Watershed ModelWatershed ModelDoug MoyerDoug Moyer1730 E. Parham Rd1730 E. Parham Rd Phone: 804-261-2634Phone: 804-261-2634Richmond, VA 23228Richmond, VA 23228 Email: dlmoyer@usgs.govEmail: dlmoyer@usgs.gov

Bacterial Source TrackingBacterial Source TrackingKen HyerKen Hyer1730 E. Parham Rd1730 E. Parham Rd Phone: 804-261-2636Phone: 804-261-2636Richmond, VA 23228Richmond, VA 23228 Email: kenhyer@usgs.govEmail: kenhyer@usgs.gov

Resources on the WebResources on the Webhttp://water.usgs.gov/owq/microbial.htmlhttp://water.usgs.gov/owq/microbial.html