U.S. Department of the InteriorU.S. Geological Survey

Bioavailability of Metals from Mining and Effects on Aquatic OrganismsChristopher J. Schmitt* U.S. Geological SurveyColumbia Environmental Research CenterColumbia, MO 65201 cjschmitt@usgs.gov

*Acknowledgements

• FWS: D. Heckathorne, D. Martin, S. Dudding, T. Nash, C. Charbonneau, G. Allen, V. Tabor, J. Dwyer

• EPA: M. Doolan• Assistance (2001) from J.

Dalgarn (BIA, Miami, OK); D. Sappington (Quapaw Tribe of OK); and A. Donahue and J. Arms (CERC)

• CERC colleagues and co-authors S. Finger, M. Wildhaber, W. Brumbaugh, T. May, J. Whyte, D.Tillitt

Spring River, Cherokee Co., KS

Spring River, Ottawa Co., OK

Pb-Zn Mining Areas in MO, OK, & KS

MO

KS

OK

From Ferderer, D.A., 1996. U.S. Geological Survey, Open-File Report 96-549

IA

IL

Old Lead Belt

(Pb/Zn; Inactive)

Tri-States District (Zn/Pb;

Inactive)

New Lead Belt

(Pb/Zn; Active)

Prospecting Area

Bioavailability Not all the metals in sediment are available

for uptake by aquatic organisms – two parts• Chemical component

• Metal association (binding) with or to particles• Assessment = sequential / partial extraction

• Particle size component• Aquatic ecosystems are particle-driven; different size

particles are utilized by organisms to differing degrees• Metals not distributed uniformly among particle sizes;

difficult to assess and often ignored• Measure uptake / accumulation by organisms

Sequential Extraction (ca. 1981)

1-N HCl(AVS-SEM)

Modified from Tessier et al. (1979, Anal Chem 51:884)

Sulfide Minerals

Pore Water

Schmitt, C.J., S.E. Finger, T.W. May, and M.S. Kaiser, 1987. Bioavailability of lead and cadmium from mine tailings to the pocketbook mussel (Lampsilis ventricosa). Pp. 115-142 in R.J. Neves, ed. Proceedings of the Workshop on Die-offs of Freshwater Mussels in the United States. U.S. Fish and Wildlife Service and Upper Mississippi River Conservation Committee. Rock Island, IL.

Big R. Pb Fractions, 1981; also Zn

Schmitt, C.J., S.E. Finger, T.W. May, and M.S. Kaiser, 1987. Bioavailability of lead and cadmium from mine tailings to the pocketbook mussel (Lampsilis ventricosa). Pp. 115-142 in R.J. Neves, ed. Proceedings of the Workshop on Die-offs of Freshwater Mussels in the United States. U.S. Fish and Wildlife Service and Upper Mississippi River Conservation Committee. Rock Island, IL.

Big R. Cd Fractions, 1981

Schmitt, C.J., S.E. Finger, T.W. May, and M.S. Kaiser, 1987. Bioavailability of lead and cadmium from mine tailings to the pocketbook mussel (Lampsilis ventricosa). Pp. 115-142 in R.J. Neves, ed. Proceedings of the Workshop on Die-offs of Freshwater Mussels in the United States. U.S. Fish and Wildlife Service and Upper Mississippi River Conservation Committee. Rock Island, IL.

Big R. Crayfish vs. Sediment Pb, 1981

Total Sediment Pb (ug/g dw)

Cra

yfis

h P

b (u

g/g

dw)

Schmitt, C.J., and S.E. Finger, 1982. The transport, fate, and effects of trace metals in the Big and Black River watersheds, Southeastern Missouri. U.S. Fish and Wildlife Service, Columbia National Fisheries Research Laboratory, Columbia, MO. Project Completion Report to the St. Louis District, U.S. Army Corps of Engineers, St. Louis, MO. 167 p.

Orconectes luteus

Uptake of Pb and Cd by LampsilisT

issu

e P

b, u

g/g

dw

Tis

sue

Cd

, ug

/g d

w

Indigenous Mussels Indigenous

Mussels

Schmitt, C.J., S.E. Finger, T.W. May, and M.S. Kaiser, 1987. Bioavailability of lead and cadmium from mine tailings to the pocketbook mussel (Lampsilis ventricosa). Pp. 115-142 in R.J. Neves, ed. Proceedings of the Workshop on Die-offs of Freshwater Mussels in the United States. U.S. Fish and Wildlife Service and Upper Mississippi River Conservation Committee. Rock Island, IL.

Pb Cd

Carcass Pb vs. Sediment Pb, 1989

Log Total Sediment Pb

Log Carcass Pb

Schmitt, C.J., M.L. Wildhaber, J.B. Hunn, T. Nash, M.N. Tieger, and B.L. Steadman, 1992. Lead in Missouri streams: Monitoring pollution from mining with an assay for δ-aminolevulinic acid dehydratase (ALA-D) in fish blood. U.S. Fish and Wildlife Service, Columbia, Missouri Field Office. 43 p.

Hypentelium nigricans

Moxostoma duquesnei

Center Creek

Big River

Blood Pb vs. Sediment Pb, 1989

Log Total Sediment Pb

Log Blood Pb

Hypentelium nigricans

Moxostoma duquesnei

Center Creek

Big River

Schmitt, C.J., M.L. Wildhaber, J.B. Hunn, T. Nash, M.N. Tieger, and B.L. Steadman, 1992. Lead in Missouri streams: Monitoring pollution from mining with an assay for δ-aminolevulinic acid dehydratase (ALA-D) in fish blood. U.S. Fish and Wildlife Service, Columbia, Missouri Field Office. 43 p.

Effects Span many levels of biological organization

• Biochemical / physiological to community• Contaminant penetrates to target sites (tissue)• Exposure pathway [Sediment-(Invertebrate?)-Fish]

Documentation = laboratory + field studies• Lab studies with field-collected sediments, spiked

sediments, pore waters, waterborne metals• Isolate effects of contaminants from other factors

• Field studies -- long-term cumulative effects• Other factors difficult to tease out – Neosho madtom• Resilience

Heme Biosynthesis and Pb

Hb Fe

Joselow, M. M. 1980. Blood zinc and lead poisoning. Pp. 171-181 in J. O. Nriagu, ed. Lead in the Environment.

-Aminolevulinic Acid Dehydratase (ALA-D) Catalyzes condensation of porphobilinogen (PBG), a

porphyrin / heme precursor, from ALA Present in most cells; greatest in blood & other heme-

rich tissues (liver, spleen, kidney, etc.) Activity inhibited by Pb; affected at Pb-B ≤0.5 mg/L

(50 µg/dL) in fish Requires Zn for activation; activity inversely correlated

w/ Zn-B in previous fish studies; reactivated by Zn (?) Activity can be measured; assay has a long history

of use as Pb biomarker in humans, birds, and fish

ALA-D Activity vs. Blood Pb in Big R. Suckers, 1981

C. J. Schmitt et al., 1984. Can. J. Fish. Aquat. Sci. 41:1030-1040.

Blood Pb (ug/g ww)

ALA

-D/H

b (n

mol

/mg/

h)

ALA-D Activity vs. Blood-Pb in Big R. Longear Sunfish, 1980

F. J. Dwyer et al., 1988. J Fish Biol 33:307-313.

Blood Pb (ug/g ww)

ALA

-D/H

b (n

mol

/mg/

h)Lepomis megalotis

ALA-D/Hb vs. Sediment Pb, 1989

Log ALA-D/Hb (nmol/mg/h

Log Total Sediment Pb (ug/g dw)

Schmitt, C.J., M.L. Wildhaber, J.B. Hunn, T. Nash, M.N. Tieger, and B.L. Steadman, 1992. Lead in Missouri streams: Monitoring pollution from mining with an assay for δ-aminolevulinic acid dehydratase (ALA-D) in fish blood. U.S. Fish and Wildlife Service, Columbia, Missouri Field Office. 43 p.

Center Creek

Big River

MO

AROK

KS

Picher Field (mines & tailings; Tribal lands)

3

Gauge

Elm Ck.

Neosho R.

Miami

6

3 mi. 5

Grand L.

Tar Ck.

Spring R.

4Gauge

2

1PicherZincville

TSMD-OK 2001

TSMD

Willow Creek

Tar Creek

Center Creek

BR / Desloge, MO

Pb >1200 µg/g dw

LB / Macon, MO

BR

LB

CERCOzCf

112 Fish / 6 Spp. / 10 Sites

SR NR REF BR

Sp 1 2 4 3(R) 5 6 (TC) 7(C) 8(O) 10(L) 9 Σ

Carp 4 4 3 4 3 5 6(b,l) 2 31

ChC 5 1 5 4 2 3 3(b) 12 35

FhC 2 2 4

LmB 2 5 4 12 1 21

SpB 3 3 1 2 9

WhC 3 2 4 3 12

Field Procedures

1

2 3

4

Laboratory Methods (Individual Fish) Age (scales only; cf not aged) Hb by azide-methemoglobin (HemoCue®) Blood metals (Fe, Pb, Cd, Zn)

• Freeeze-dried and digested in tube (clean!)• Analyzed by ICPMS

Liver (carp only) and carcass metals (Pb, Cd, Zn; not LB) by ICPMS (dw liver only)

ALA-D by microtiter plate method (modified from Granick 1972 and Hodson 1976; described by Whyte et al. 2002)

PBG Standard Curve (X 3)

Blank (X 3)

ALA (X 3))

ALA-D Assay, 96-Well Microtiter Plate (12 samples + 9 standards)

Color results from Ehrlich’s reagent binding with PBG

Plate is scanned with an automated plate reader

Nmol PBG/µL blood/h

Nmol PBG/mg Hb/h

Si t e 1 2 3 45 6 9 10

l pbbd

- 1

0

1

2

l pbl d

- 1. 0 - 0. 8 - 0. 6 - 0. 4 - 0. 2 0. 0 0. 2 0. 4 0. 6 0. 8

BR

LBSite 6

(NR-TC)

Site 6(NR-TC)

Pb

-B (

ug

/g d

w)

NAWQA carp range (0.05-2.20 μg/g dw, n = 182)

5.4 μ/g (n = 1)

Pb-B vs. Pb-L in Carp, 2001*

Pb-L (ug/g dw)

y = x

Site 3(NR-Ref)

n = 30, r = 0.88, p < 0.01

*2002 for LB

Pb*, Cd*, and Zn* in Carp, 2001**

W. G. Brumbaugh et al. 2005, Arch Environ Contam Toxicol (in press).

0.1

1

10

110

1000.1

1

10

Liv

er

Blood

Car

cass

Pb

0.1

1

10

100

1000

0.010.1

10.01

0.1

1

Liv

erBlood

Car

cass

Cd

100

1000

10000

100200

300

100

200

300

Liv

er

Zn

* Blood, liver and carcass; all ug/g dw

** 2002 for LB

R2 = 0.78 – 0.88 (L), 0.49 – 0.72 (C)

R2 = 0.69 (L), 0.61 (C)

NS

SR & NRDischarge,Jun-Nov 2001

Collections originally scheduled for July ’01

Postponed until October due to unusually high flows

High flow may have induced carp to move

Site 3 (SR)

Site 4 (NR)

Speci es Car p ChC FhCLmB SpB WhC

l al ad

- 0. 4

- 0. 3

- 0. 2

- 0. 1

0. 0

0. 1

0. 2

0. 3

0. 4

0. 5

0. 6

0. 7

0. 8

0. 9

1. 0

1. 1

l pbbw

- 3 - 2 - 1 0 1

ALA-D/Hb vs. Pb-B, 2001*

Pb-B (ug/g ww)

All MicropterusALA-D/Hb = 0.258 – 0.219 Pb-B ,

n = 29, r2 = 0.68, p <0.01

BR

AL

A-D

/Hb

(n

mo

l/mg

/h)

All catfish (mostly ChC)ALA-D/Hb = -1.206 – 0.237 Pb-B +

0.868 Zn-B, n = 35, R2 =0.65

C. J. Schmitt et al., 2005. Environ Toxicol Chem 24, in press. *2002 for LB

Speci es Car p ChC FhCLmB SpB WhC

l f ebw

2. 0

2. 1

2. 2

2. 3

2. 4

2. 5

2. 6

2. 7

l pbbw

- 3 - 2 - 1 0 1

Fe-B (or Hb) vs. Pb-B, 2001*

BR

Carp

Pb-B (ug/g ww)

LB

CERC

Fe-

B (

ug

/g w

w)

C. J. Schmitt et al., 2005. Environ Toxicol Chem 24, in press. *2002 for LB

Summary (Bioavailability) Contamination by metals evident in TSMD fish

• Pb concentrations greater than background, but lower than in other historic mining areas (BR, etc.)

• Metals bioavailability issue resolved (blood, liver metals; correlations)• Penetration to active sites / tissues• Redundancy / QA vs. efficiency

• Variable results for carp suggest fish movement, possibly induced by flooding

ALA-D activity differed among sites (some taxa) • Relationship vs Pb-B well documented in catfish and

bass, less so in carp• Differences among taxa apparent• Sensitivity of ictalurids re. Neosho madtom?

• Bioavailability, unique biochemical response • No detectable effects on Hb or Fe-B• Exposure indicator*

Higher level effects also evident (fish, invertebrates; dns)

Future studies: Aquatic effects of Cd, Zn, and ?

Summary (Effects)