Antibiotics in Our Streams

Antibiotics in Our StreamsAntibiotics in Our StreamsHow Much is There and Where Are They Coming From?How Much is There and Where Are They Coming From?

Colorado State UniversityColorado State University

Kenneth CarlsonKenneth Carlson

(([email protected]@engr.colostate.edu))

SungSung--Chul KimChul KimShinwooShinwoo YangYang

October, 2005October, 2005

Annual Antimicrobials UsageAnnual Antimicrobials Usage2002 Animal Health Industry Sales

Total Pounds Total Pounds

(Thousands)(Thousands)Percent of Total (%)Percent of Total (%)

Human UsesHuman Uses

Disease TreatmentDisease Treatment

Other Human UseOther Human Use

3,0003,000

1,5001,50099

55

Livestock UsesLivestock Uses

Therapeutic Therapeutic –– All SpeciesAll Species

Nontherapeutic Nontherapeutic –– Cattle, Swine, Poultry Cattle, Swine, Poultry

2,0002,000

24,50024,50066

7777

OthersOthers

Companion AnimalsCompanion Animals 1,0001,000 33

Total Antimicrobials UseTotal Antimicrobials Use 32,00032,000 100100

Sources, Distribution and Sinks of Sources, Distribution and Sinks of Pharmaceuticals in the EnvironmentPharmaceuticals in the Environment

Sources from: K Kummerer, Pharmaceuticals in the Environment, SpSources from: K Kummerer, Pharmaceuticals in the Environment, Springerringer

Human Used Pharmaceuticals:Human Used Pharmaceuticals:STP as point sourceSTP as point source

Veterinary Pharmaceuticals:Veterinary Pharmaceuticals:Manure as fertilizer Manure as fertilizer ⇒⇒ Soil Soil ⇒⇒Surface Runoff or Surface Runoff or Leaching through subLeaching through sub--surfacesurface

Risk to Drinking Water andRisk to Drinking Water andWatershed EcologyWatershed Ecology

Main ObjectivesMain Objectives

Monitoring Occurrence of Human and Animal Monitoring Occurrence of Human and Animal

Antibiotics in Water and SedimentAntibiotics in Water and Sediment

Monitoring Occurrence in Animal Waste Lagoons and Monitoring Occurrence in Animal Waste Lagoons and ManureManure

Evaluating Transport Pathways and MechanismsEvaluating Transport Pathways and Mechanisms

Evaluating Fate of Antibiotics and Waste Management Evaluating Fate of Antibiotics and Waste Management

Practice (BMPs)Practice (BMPs)

NH2

OHO

O OHOH

O

OH

NCH3 CH3R1R2

R3R4

12

34

56

78

91011

1214

1516

17ABCD

13

pKaaCompounds R1 R2 R3 R4 pK1 pK2 pK3

Oxytetracycline(OTC) OH OH CH3 H 3.3 7.3 9.1

Chlortetracycline(CTC) H OH Cl

3.3 7.4

9.3

Minocycline(MNC) H H H 2.8 5.0 7.8Demeclocycline(DMC) H OH H Cl

Melocycline(MCC) OH CH2 - ClTetracycline(TC) H OH CH3 H 3.3 7.7 9.7

Doxycycline(DXC) OH H CH3 H 3.5 7.7 9.5

TetracyclinesTetracyclines

Name

R Name R pKa

Sulfathiazole(STZ)

S

N

Sulfametho-xazole(SMX)

ON

CH3

Sulfamerazine(SMR)

N

N

CH3

Sulfachloro-

pyridazine

(SCP)

NN Cl

Sulfamethazine(SMT)

N

N

CH3

CH3

Sulfadimeth-

oxine

(SDM)N

N

CH3O

CH3O

NH2

SO

O

NH

R

SulfonamidesSulfonamides

MacrolidesMacrolides

H

CH3 H

OH

O

CH3

O

CH3

CH3

O

CH3

O

OHCH3

OOH

CH3

OH OCH3

OCH3

OHCH3

NCH3CH3

C

OCH3

H

CH3 H

OH

O

CH3

H3CO-H2C-H2C-O-H2C-O-N

CH3

CH3

O

CH3

O

OHCH3

OOH

CH3

OH OCH3

OCH3

OHCH3

NCH3CH3

C

OCH3

O

C2H3 O

CH3

O

CH3OH

O

CH3

O

O CH3

CHOOH O

NCH3 CH3

OOH

CH3

OH

CH3

OHO

CH3

OCH3OCH3

Erythromycin

Tylosin

Roxithromycin

IonophoresIonophores

HCH3

O

O

OH

O O

O

H5C2

HCH3

CH3

HO

OH

OH

CH3CH3OCH3

H

CH3

CH3

OHCH3

H3C

1

234

5

6

78

91 0

1 1

1 2 1 3

1 4

1 5

1 6

1 7 1 81 9

2 0

2 12 2

2 3

2 4

2 5

2 6

2 7

2 82 9

HCH3

O

O

OH

O O

O

H5C2

HCH3

CH3

HO

OH

OH

CH3CH3O

H

CH3

CH3

OHCH3

H3C

1

23

4

5

6

78

910

11

12 13

14

15

16

17 1819

20

2122

23

24

25

26

27

2829

CH3OH3C

CH3

H

O

OO

OH CH3

CH3

H

O

OH

O

CH3

H

OHOH

CH3

CH3

CH3 CH2OH

12

3

4

56

78

9

10 11

12 13

14 15

16 17

18 19

20

2122

23

2425

Monensin

Salinomycin Naracin

Main ObjectivesMain Objectives

Monitoring Occurrence of Human and Animal Monitoring Occurrence of Human and Animal

Antibiotics in Water and SedimentAntibiotics in Water and Sediment

Monitoring Occurrence in Animal Waste Lagoons and Monitoring Occurrence in Animal Waste Lagoons and ManureManure

Evaluating Transport Pathways and MechanismsEvaluating Transport Pathways and Mechanisms

Evaluating Fate of Antibiotics and Waste Management Evaluating Fate of Antibiotics and Waste Management

PracticePractice

Solid Phase Extraction (SPE) ProcedureSolid Phase Extraction (SPE) Procedure

HLBHLB3mL MeOH

3mL HCL(5%,v/v)3mL DI

3mL MeOH3mL HCL(5%,v/v)

3mL DIFlow Rate2mL/min

Flow Rate2mL/min

3×3mLDI Water3×3mL

DI Water2×2.5mLMeOH

2×2.5mLMeOH

Schematic Diagram of the Sediment Sample PreparationSchematic Diagram of the Sediment Sample Preparation

SampleSample(Air dried)(Air dried)

PrePre--ExtractionExtraction

CleanClean--upup(SPE)(SPE)

Evaporation and ReconstitutionEvaporation and Reconstitution

HPLC/MS/MS AnalysisHPLC/MS/MS Analysis

TCsTCs and and SAsSAs::Mcllvaine Buffer Solution (pH 4.0)Mcllvaine Buffer Solution (pH 4.0)

MLsMLs and and IPsIPs::Ammonium Hydroxide Buffer (pH 10.0)Ammonium Hydroxide Buffer (pH 10.0)

Nitrogen Gas Water Bath (50Nitrogen Gas Water Bath (50°°C)C)5050μμl Sample + 70l Sample + 70μμl mobile phase Al mobile phase A

High Performance Liquid Chromatography Tandem High Performance Liquid Chromatography Tandem Mass Spectrometry (HPLC/MS/MS)Mass Spectrometry (HPLC/MS/MS)

EquipmentEquipment HP 1100 HPLC equipped with HP 1100 HPLC equipped with ThermostattedThermostatted Auto Sampler and Auto Sampler and

variable UV detectorvariable UV detector

ThermoFinniganThermoFinnigan LCQ Duo ion trap mass spectrometerLCQ Duo ion trap mass spectrometer

XterraXterra MS CMS C1818 (2.1(2.1××50mm, 2.550mm, 2.5μμm pore size, endm pore size, end--capped)capped)

Optimized HPLCOptimized HPLC

ConditionConditionColumnColumn

TemperatureTemperature

((°°C)C)

Flow RateFlow Rate

(ml/min)(ml/min)Mobile Phase Conditions:Mobile Phase Conditions:

Mobile Phase A (99.9% DI+ 0.1% Formic Acid)Mobile Phase A (99.9% DI+ 0.1% Formic Acid)

Mobile Phase B (99.9% ACN + 0.1% Formic Acid)Mobile Phase B (99.9% ACN + 0.1% Formic Acid)

Mobile Phase C (100% Mobile Phase C (100% MeOHMeOH))

Tetracyclines (Tetracyclines (TCsTCs))

Sulfonamides (Sulfonamides (SAsSAs))1515 0.320.32 A: 96% + B: 4%: 0 (min) A: 96% + B: 4%: 0 (min) ⇒⇒ A: 70% + B: 30%: 29 (min) A: 70% + B: 30%: 29 (min) ⇒⇒

A: 96% + B: 4%: 30 (min) A: 96% + B: 4%: 30 (min)

Macrolides (Macrolides (MLsMLs)) 4545 0.320.32 A: 80% + B: 20%: 0 (min) A: 80% + B: 20%: 0 (min) ⇒⇒ A: 65% + B: 35%: 13 (min) A: 65% + B: 35%: 13 (min) ⇒⇒A: 80% + B: 20%: 14 (min) A: 80% + B: 20%: 14 (min)

Ionophore PolyethersIonophore Polyethers

((IPsIPs))1515 0.250.25 A: 50% + C: 50%: 0 (min) A: 50% + C: 50%: 0 (min) ⇒⇒ A: 10% + C: 90%: 1 (min) A: 10% + C: 90%: 1 (min) ⇒⇒

A: 50% + C: 50%: 20 (min) A: 50% + C: 50%: 20 (min)

Optimized MSOptimized MS

ConditionConditionNitrogen Gas used for drying and Nitrogen Gas used for drying and nebulizingnebulizing

Spray Voltage Spray Voltage –– 4.5V4.5V

Capillary Voltage Capillary Voltage –– 21V21V

Capillary Temperature Capillary Temperature -- 165165°°CC

Quality Assurance: Quality Assurance: Recovery,Limit of Quantification (LOQ)Recovery,Limit of Quantification (LOQ)

RecoveryRecovery

Range (%)Range (%)TCsTCs SAsSAs MLsMLs IPsIPs

AqueousAqueous

(0.1,1,5 (0.1,1,5 μμg/L)g/L)

102 102 -- 124124 77 77 –– 124124 89 89 –– 114114 82 82 –– 117117

SedimentSediment

(1,30,90 (1,30,90 μμg/kg)g/kg)

33 33 --115115 62 62 -- 108108 53 53 -- 128128 51 51 -- 105105

Lowest calibration curve point based on signal/noise 3 (0.01 Lowest calibration curve point based on signal/noise 3 (0.01 μμg/L and 1 g/L and 1 μμg/kg )g/kg )Standard Deviation Standard Deviation ×× StudentStudent’’s ts t--variate for one sided variate for one sided tt--test at the 95% confidence intervaltest at the 95% confidence interval

LOQLOQ TCsTCs SAsSAs MLsMLs IPsIPs

AqueousAqueous

((μμg/L)g/L)

0.01 0.01 –– 0.040.04 0.01 0.01 –– 0.020.02 0.01 0.01 –– 0.030.03 0.012 0.012 –– 0.0270.027

SedimentSediment

((μμg/kg)g/kg)

0.3 0.3 –– 2.52.5 0.3 0.3 –– 1.71.7 0.5 0.5 –– 1.71.7 0.4 0.4 –– 3.63.6

Detect Response of Spiked Sample after SPEDetect Response of Spiked Sample after SPE

Detect Response of Spiked Sample prior to SPEDetect Response of Spiked Sample prior to SPE×× 100100Recovery Ratio =Recovery Ratio =

Sampling Sites DescriptionSampling Sites DescriptionSampling Site 1:Sampling Site 1:

Pristine watershedPristine watershed

Sampling Site 2:Sampling Site 2:

Rural landscapeRural landscape


Urban influencedUrban influenced

Wastewater Reclamation FacilityWastewater Reclamation Facility


Heavily agricultural influenced areaHeavily agricultural influenced area


Both urban and agricultural Both urban and agricultural

influenced area influenced area

+ Sampling Sites • Dairy ■ Feedlots ▲ Ranches ★USGS Gages

Site 2 Site 2 -- AqueousAqueous

Site 2

Tetracyclines

TC CTC OTC DMC MCC DXC

Con

cent

ratio

n ( μ

g/L)

0.00

0.01

0.02

0.03

0.04

0.05

0.06

May-03April-04August-04February-05

Site 2

Sulfonamides

STZ SMR SMT SCP SMX SDM

Con

cent

ratio

n ( μ

g/L)

0.00

0.01

0.02

0.03

0.04

0.05


Site 2

Macrolides

ETM-H2O RTM TLS

Con

cent

ratio

n ( μ

g/L)

0.00

0.01

0.02

0.03

0.04


Site 2

Ionophore Polyethers

MNS SLM NRS

Con

cent

ratio

n ( μ

g/L)

0.000

0.005

0.010

0.015

0.020

0.025

0.030


No observedNo observedconcentrationconcentrationin May and Aprilin May and Aprilexcept except NarasinNarasin

Local small farmLocal small farmmight contributemight contributethe detection of the detection of NarasinNarasin

Higher concentrationHigher concentrationin February within February withhigh detect frequencyhigh detect frequencyFor For TCsTCs and and SAsSAs


Site 3

Tetracyclines


Con

cent

ratio

n (μ

g/L)

0.00

0.02

0.04

0.06

0.08

0.10

0.12


Site 3

Sulfonamides

STZ SMR SMT SCP SMX SDM

Con

cent

ratio

n ( μ

g/L)

0.00

0.05

0.10

0.15

0.20

0.25

0.30

0.35

Site 3

Macrolides

ETM-H2O RTM TLS

Con

cent

ratio

n ( μ

g/L)

0.0

0.1

0.2

0.3

0.4

0.5


Site 3


MNS SLM NRS

Con

cent

ratio

n ( μ

g/L)

0.00

0.01

0.02

0.03

0.04


Higher concentrationHigher concentrationof human usedof human usedantibiotics orantibiotics orantimicrobialsantimicrobials

High concentration ofHigh concentration ofETMETM--H2O in low flowH2O in low flowand cold temperatureand cold temperature


Site 4

Tetracyclines


Con

cent

ratio

n ( μ

g/L)

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4


Site 4

Sulfonamides

STZ SMR SMT SCP SMX SDMC

once

ntra

tion

( μg/

L)0.00

0.02

0.04

0.06

0.08

Site 4

Macrolides

ETM-H2O RTM TLS

Con

cent

ratio

n ( μ

g/L)

0.00

0.02

0.04

0.06

0.08

0.10


Site 4


MNS SLM NRS

Con

cent

ratio

n ( μ

g/L)

0.000

0.005

0.010

0.015

0.020

0.025

0.030

0.035


3 3 IPsIPs were detectedwere detected

Site 5 Site 5 -- AqueousAqueousSite 5

Tetracyclines


Con

cent

ratio

n ( μ

g/L)

0.0

0.1

0.2

0.3

0.4

Site 5

Sulfonamides


once

ntra

tion

( μg/

L)0.00

0.02

0.04

0.06

0.08

0.10

0.12

0.14

0.16

Site 5

Macrolides

ETM-H2O RTM TLS

Con

cent

ratio

n ( μ

g/L)

0.00

0.05

0.10

0.15

0.20

0.25


Site 5


MNS SLM NRS

Con

cent

ratio

n ( μ

g/L)

0.00

0.01

0.02

0.03

0.04


Higher concentrationHigher concentrationof MNS in Mayof MNS in May

Site 2 Site 2 -- SedimentSedimentSite 2

Tetracyclines


Con

cent

ratio

n ( μ

g/kg

)

0

10

20

30

40

50


Site 2

Sulfonamides


once

ntra

tion

( μg/

kg)

0

1

2

3

4

5

6

7


Site 2

Macrolides

ETM-H2O RTM TLS

Con

cent

ratio

n ( μ

g/kg

)

0.00

2.00

4.00

6.00

8.00

10.00

12.00

14.00


Site 2


MNS SLM NRS

Con

cent

ratio

n ( μ

g/kg

)

0.0

0.5

1.0

1.5

2.0

2.5

3.0


No measurement forNo measurement forMLsMLs and and IPsIPsin Februaryin February

More frequent detectionMore frequent detectioncompared to aqueouscompared to aqueous


Tetracyclines


Con

cent

ratio

n ( μ

g/kg

)

0

20

40

60

80

100

120


Site 3

Sulfonamides


once

ntra

tion

( μg/

kg)

0

2

4

6

8

Site 3

Macrolides

ETM-H2O RTM TLS

Con

cent

ratio

n ( μ

g/kg

)

0

5

10

15

20

25

30


Site 3


MNS SLM NRS

Con

cent

ratio

n ( μ

g/kg

)

0

1

2

3

4


Higher concentrationHigher concentrationIn February for In February for TCsTCs andandSAsSAs

No No IPsIPs except except NarasinNarasin


Tetracyclines


Con

cent

ratio

n ( μ

g/kg

)

0

5

10

15

20

25

30


Site 4

Sulfonamides


once

ntra

tion

( μg/

kg)

0

2

4

6

8

10

12

14

16

Site 4

Macrolides

ETM-H2O RTM TLS

Con

cent

ratio

n ( μ

g/kg

)

0

2

4

6

8

10

12

14

16

18


Site 4


MNS SLM NRS

Con

cent

ratio

n ( μ

g/kg

)

0

5

10

15

20

25

30

35


Higher concentrationHigher concentrationin August for SMT,in August for SMT,ETMETM--HH22O, and O, and IPsIPs

High concentrationHigh concentrationOf CTC, STZ, and TLSOf CTC, STZ, and TLS


Tetracyclines


Con

cent

ratio

n ( μ

g/kg

)

0

20

40

60

80

Site 5

Sulfonamides


once

ntra

tion

( μg/

kg)

0

2

4

6

8

Site 5

Macrolides

ETM-H2O RTM TLS

Con

cent

ratio

n ( μ

g/kg

)

0

2

4

6

8

10

12

14


Site 5


MNS SLM NRS

Con

cent

ratio

n ( μ

g/kg

)

0

5

10

15

20

25

30

35


Combination of human usedCombination of human usedAnd veterinary medicinesAnd veterinary medicines

Summary of Measured Concentration at watershedSummary of Measured Concentration at watershedWaterWater SedimentSediment

Detected Concentration (Detected Concentration (μμg/L)g/L) Detected Concentration (Detected Concentration (μμg/kg)g/kg)CompoundsCompounds Frequency of Frequency of

Detection (%)Detection (%) MeanMean Standard Standard DeviationDeviation

MaximumMaximum MinimumMinimum

Frequency of Frequency of

Detection (%)Detection (%) MeanMean Standard Standard DeviationDeviation

MaximumMaximum MinimumMinimum

TCTC 4242 0.020.02 0.010.01 0.030.03 0.010.01 8080 17.917.9 24.324.3 102.7102.7 1.11.1

CTCCTC 4545 0.080.08 0.070.07 0.210.21 0.010.01 8080 10.810.8 9.29.2 30.830.8 1.11.1

OTCOTC 3737 0.180.18 0.400.40 1.211.21 0.010.01 8080 14.814.8 13.913.9 56.156.1 2.42.4

DMCDMC 3030 0.030.03 0.010.01 0.050.05 0.020.02 8080 6.96.9 5.55.5 23.623.6 2.12.1

MCCMCC 4747 0.030.03 0.030.03 0.100.10 0.010.01 8080 24.324.3 21.521.5 72.072.0 4.34.3

DXCDXC 3333 0.020.02 0.010.01 0.050.05 0.010.01 6565 15.715.7 10.110.1 38.938.9 2.22.2

STZSTZ 3333 0.010.01 0.010.01 0.030.03 0.010.01 7070 3.33.3 2.32.3 5.45.4 1.31.3

SMRSMR 3030 0.020.02 0.020.02 0.060.06 0.010.01 1515 4.84.8 2.32.3 6.86.8 2.32.3

SMTSMT 1010 0.020.02 0.000.00 0.020.02 0.020.02 2525 4.74.7 5.25.2 13.713.7 1.01.0

SCPSCP 55 0.030.03 0.000.00 0.030.03 0.030.03 2525 2.72.7 0.50.5 3.23.2 1.91.9

SMXSMX 6060 0.110.11 0.090.09 0.320.32 0.040.04 2525 1.61.6 0.30.3 1.91.9 1.21.2

SDMSDM 3333 0.020.02 0.020.02 0.040.04 0.010.01 3030 3.83.8 2.12.1 6.86.8 1.71.7

ETMETM--H2OH2O 6565 0.120.12 0.130.13 0.450.45 0.020.02 7575 10.010.0 7.67.6 25.625.6 1.31.3

RTMRTM 00 0.000.00 0.000.00 0.000.00 0.000.00 3030 2.12.1 1.91.9 5.95.9 1.11.1

TYLTYL 55 0.050.05 0.000.00 0.050.05 0.050.05 5353 3.03.0 2.82.8 9.39.3 1.11.1

MNSMNS 5050 0.0090.009 0.0010.001 0.0360.036 0.0020.002 4040 13.813.8 11.211.2 31.531.5 2.42.4

SLMSLM 4343 0.0020.002 0.0020.002 0.0070.007 0.0010.001 4040 9.19.1 11.011.0 30.130.1 1.81.8

NRSNRS 3737 0.0180.018 0.0160.016 0.0380.038 0.0010.001 4040 6.46.4 4.24.2 16.316.3 2.82.8

Pseudo Partitioning CoefficientPseudo Partitioning Coefficient

Sediment Concentration (Sediment Concentration (μμg/kg)g/kg)Pseudo Partitioning Coefficient (L/kg) =Pseudo Partitioning Coefficient (L/kg) =

Aqueous Concentration (Aqueous Concentration (μμg/L)g/L)

CompoundsCompounds RangeRange AverageAverage ReferenceReference

CTCCTC 623 623 -- 11601160 891891

2424

243243

15251525

670 670 –– 16201620

SMXSMX 20 20 -- 2828 0.6 0.6 –– 4.94.9

ETMETM--H2OH2O 72 72 -- 397397 8.3 8.3 –– 128128

MNSMNS 1000 1000 -- 20502050 --

PPC: MNS > CTC > ETMPPC: MNS > CTC > ETM--H2O > SMXH2O > SMX

Sources from: J Tolls, Environmental Science and Technology (200Sources from: J Tolls, Environmental Science and Technology (2001), 35, 3397 1), 35, 3397 -- 34063406

Animal Waste OperationsAnimal Waste Operations

Operation Lagoon Water Manure

Dairy 7 7

Beef 6 5

Hog 10 5

Sheep 2 4

Turkey 0 2

Animal Waste OperationsAnimal Waste OperationsConcentration in lagoon water

Samples OTC CTC TC MNSug/L ug/L ug/L ug/L

MDL 1.79 1.33 2.09 0.86

Dairy A 5.04 1.71 9.89Dairy B 17.46 4.78 11.48Dairy C 10.62 1.56Dairy D 2.25Dairy EDairy F 2.95 12.45Dairy G 2.41Beef A 3.27 51.80 7.44 101.34Beef B 7.85 54.56Beef C 2.08 6.94 502.34Beef D 3.53 3.02 8.61 75.32Beef E 2.31 43.27Beef F 2.52 395.76Hog A 7.53 12.93Hog B 2.36 6.23Hog C 11.30 1193.00 6.47Hog D 12.77 2.54Hog E 135.43 16.95Hog F 99.52 14.40Hog G 1524.82 9498.69 26.47Hog H 1310.23 9450.37 24.83Hog I 6.65 2.58Hog J 8.05 2.40

Sheep 1 1.60 6.29Sheep 2


Samples STZ SCP SDM SMR SMT SMXug/L ug/L ug/L ug/L ug/L ug/L

MDL 0.81 0.87 1.03 1.02 0.94 0.58

Dairy A 3.83 4.41 0.95 0.61Dairy B 1.01 16.80 1.02 2.91 0.59Dairy C 1.76 2.34Dairy DDairy EDairy F 28.04Dairy G 1.82Beef A 1.44 2.43 0.64Beef B 1.17Beef CBeef D 3.83Beef E 2.14 0.96 0.70Hog A 2.07Hog B 4.43Hog C 1941.53 66.07Hog D 1.77Hog E 3.80 2.96 1.12Hog F 3.88 4.53 2.70 1.26Hog G 2079.27 270.73 1.59 9.44Hog H 2430.30 280.58 2.31 8.05Hog IHog J 1.51 1.19

Sheep 1 64.23 2.47 0.60Sheep 2 132.68


Samples TLS ETM-H2O RTMug/L ug/L ug/L

MDL 0.46 0.65 0.59

Dairy A 1.16Dairy B 4.05 3.05 19.20Dairy C 0.51Dairy D 9.60 0.74Dairy E 19.53 7.95 4.58Dairy F 5.88Dairy G 4.84Beef A 102.52 7.39 43.66Beef B 0.90 0.68Beef C 11.69Beef D 27.88Beef E 37.59Beef F 36.78 0.67Hog A 0.69 0.63Hog BHog C 12.58 3.76 1.09Hog D 60.01 4.25Hog E 5.04Hog F 2.27Hog G 0.46Hog HHog IHog J

Sheep 1Sheep 2 1.80 1.40 1.06

Animal Waste OperationsAnimal Waste OperationsConcentration in liquid manure

Samples OTC CTC TC MNSug/kg ug/kg ug/kg ug/kg

MDL 2.43 1.93 2.27 1.96

Dairy/Fresh 16.32 8.87 45.00Dairy/Fresh 42.43 33.38Dairy/Fresh 38.62 34.56Dairy/Fresh 15.73 356.62Dairy/Fresh 5130.42 2.47 1045.52Dairy/Fresh 5.58 39.01 78.53Dairy/Fresh 620.92 31.34 83.00Beef/Fresh 4.58 345.67Beef/Fresh 20.59 380.77 585.34 542.57Beef/Fresh 6.05 10.28 983.56Beef/Fresh 22.93 234.49 184.97 456.30Beef/Fresh 8.29 30.71 36.78Hog Fresh 11.94 23.88 128.44Hog Fresh 91.73 23138.27 1115.56Hog Fresh 1076.27 634.89 394.06Hog Fresh 372.78 148.60 284.15Hog Fresh 176779.21 22837.04 808.12

Sheep Fresh 172.33 3622.96 343.53Sheep Fresh 5.90 5.07 26.45Sheep Fresh 34.04 9990.72 3071.49Sheep Fresh 175832.67 9.64 10896.72Turkey/Old 160.53 26.91Turkey/Old 18.69 308.70 37.78

Animal Waste OperationsAnimal Waste OperationsActual concentration in liquid manure

Samples STZ SCP SDM SMR SMT SMXug/kg ug/kg ug/kg ug/kg ug/kg ug/kg

MDL 1.35 1.43 1.54 1.17 1.62 1.08

Dairy/Fresh 16.51 7.85 1.75Dairy/Fresh 7.04 1.67Dairy/Fresh 11.49 1.89Dairy/Fresh 8.57Dairy/Fresh 24.32 3.04 647.85 2.64 3.57Dairy/Fresh 46.39 9.53 3.01 1.47Dairy/Fresh 2.74 11.67Beef/Fresh 12.77 3.25Beef/Fresh 127.47 2.18 12.11 5.30 7.28Beef/Fresh 19.70 6.31 2.82Beef/Fresh 257.78 8.43 2.49Beef/Fresh 7.88 5.97Hog Fresh 14.75 1.72 3.07 3.59Hog Fresh 17.40 3.19 2.71Hog Fresh 4.76 5.73Hog Fresh 36.69 38.38 1.81 1.50Hog Fresh 1.74

Sheep Fresh 1.37Sheep Fresh 12.96 419.21 1.96Sheep Fresh 4.19 1.82 2.01Sheep Fresh 10.19 248.42 2.73 1.80Turkey/Old 31.35 69.86 3.18 6.47Turkey/Old 5.51 6.17 1.68

Column Leaching ExperimentsColumn Leaching Experiments

PurposesPurposesEvaluating potential mobility of antibioticsEvaluating potential mobility of antibiotics

Estimating partition coefficient (Estimating partition coefficient (KdKd))

Comparing surface runoff field experimentComparing surface runoff field experiment

Expected ResultsExpected ResultsMobility: Sulfonamides > Mobility: Sulfonamides > MacrolidesMacrolides > > IonophoresIonophores > >

TetracyclinesTetracyclines

KdKd values: values: TetracyclinesTetracyclines > > IonophoresIonophores > > MacrolidesMacrolides > > SulfonamidesSulfonamides

Mass Loading From Local Irrigation Ditches Mass Loading From Local Irrigation Ditches to Watershedto Watershed ID Site name or

descriptionAveraged MeanStream Flow (cfs) a

A Eaton Canal 195

B Local Ditch -

C Eaton Canal 195

D Graham Seep Ditch 3

E Greeley #2 Canal 119

F Graham Seep Ditch 3

G Eaton Canal 195

H Greeley #2 Canal 119

I Greeley #2 Canal 119

M Agricultural Area 45

N Urban and agricultural combined area

a Averaged mean stream flow of canal or ditch was provided by Cache la Poudre River water commitionerand river mean stream flow was adapted from U.S. Geological Survey (USGS). Unknown data is not shown.

Inflow Inflow TC: 153.4TC: 153.4

CTC: 266.4CTC: 266.4OutflowOutflowTC: 2.5TC: 2.5

CTC: 2.9CTC: 2.9Mass Flux Mass Flux

TC: 1.6TC: 1.6CTC: 1.1CTC: 1.1

Inflow Inflow STZ: 6.2STZ: 6.2

SMT: 151.3SMT: 151.3OutflowOutflowSTZ: 0.3STZ: 0.3SMT: 3.0SMT: 3.0

Mass Flux Mass Flux STZ: 4.6STZ: 4.6

SMT: 2.0SMT: 2.0

Inflow Inflow ETMETM--HH22O: 48.2O: 48.2

TLS: 118.3TLS: 118.3OutflowOutflow

ETMETM--HH22O: 2.1O: 2.1TLS: 4.2TLS: 4.2

Mass FluxMass FluxETMETM--HH22O: 4.4O: 4.4

TLS: 3.5TLS: 3.5

InflowInflowMNS: 13.0MNS: 13.0SLM: 6.0SLM: 6.0NRS: 12.6NRS: 12.6OutflowOutflowMNS: 0.3MNS: 0.3SLM: 0.3SLM: 0.3NRS: 0.6NRS: 0.6

Mass FluxMass FluxMNS: 1.9MNS: 1.9SLM: 5.0SLM: 5.0NRS: 4.5NRS: 4.5

Unit of Inflow and Outflow: ton/yr, Mass Flux: %Unit of Inflow and Outflow: ton/yr, Mass Flux: %

ConclusionsConclusions

•• Occurrence frequency high in semiOccurrence frequency high in semi--arid watershedarid watershed

•• SedimentSediment--partitioned concentrations much higherpartitioned concentrations much higher

•• With different site characteristics and land use, With different site characteristics and land use, concentration of human and animal antibiotics varies concentration of human and animal antibiotics varies significantly significantly –– BMPs can be developedBMPs can be developed

•• Surface runoff may contribute to the release of Surface runoff may contribute to the release of antibiotics or antimicrobials to the environmentantibiotics or antimicrobials to the environment

•• Local irrigation ditches might act as intermediate Local irrigation ditches might act as intermediate transport pathwaystransport pathways

AcknowledgementAcknowledgement

Colorado State UniversityColorado State University

Dr Amy Dr Amy PrudenPruden

Dr Jessica DavisDr Jessica Davis

Kathy Kathy DoeskenDoesken

Support:Support:

United States Department of Agriculture (USDA)United States Department of Agriculture (USDA)

National Integrated Water Quality ProgramNational Integrated Water Quality ProgramIntegrated Research, Education, and Extension Competitive GrantsIntegrated Research, Education, and Extension Competitive Grants ProgramProgram

Documents

Antibiotics in Our Streams