9th Annual Ronald C. Baird Sea Grant Science Symposium

Environmental Contaminants in Shell Diseased vs. Non-Diseased American

Lobsters (Homarus americanus)

Lawrence A. LeBlanc, Deanna L. Prince and Hans Laufer

School of Marine Sciences,Department of Food Science and Human Nutrition

University of Maine, Orono, MEDepartment of Molecular and Cell Biology, University of Connecticut,

Storrs, CT

Is contaminant exposure correlated to the presence of lobster shell disease?

Metals & Organics have a variety of toxic effects:– neurotoxicity, endocrine disruption, narcotic

toxicity

– Little is known of the interaction effects of metal and organic contaminants

– A combination of stressors may lead to compromised immune systems

Why Shell Disease and Contaminants?

• Shell disease in crustaceans more prevalent in impacted environments– Lobster- Massachusetts Bay,

Buzzards Bay (Estrella, 1984)

• Evidence of elevated concentrations of trace metals with shell disease – Blue Crabs (Weinstein, et al.

1992)– Lobster (Vassiliev, 2004)

• Alkylphenols are postulated to have a role in shell disease (Laufer et al.)

Two Approaches

• LeBlanc and Prince:– Compare patterns of trace metal and organic

contaminant accumulation in shell-diseased and non-diseased lobsters

• Laufer et al.– Investigations of the effects of alkylphenol

compounds on shell thinning

Our Hypothesis:

• There is a relationship between epizootic shell disease and contaminant body burdens

Project Objectives• Determine whether there is a relationship between

contaminant concentrations in hepatopancreas/hemolymph and lobster shell disease

• Evaluate the use of lobster hemolymph as a rapid screening tool for contaminant exposure

• Develop a multiresidue method for a wide suite of organic contaminants suitable for lobster hepatopancreas and hemolymph tissues”

• Examine the partitioning of metals between various component tissues (including gills, ovaries, muscle, exoskeleton)

Project Objectives

• Why Hepatopancreas?– Concentrations of many trace

elements are higher than in muscle tissue

– Elevated concentrations of lipophilic organic contaminants

• Why Hemolymph?– Easier matrix to work with

Less labor-intensive sample preparation

Lobster Collection• Lobster collections:

– Regional: RI, LIS, ME– Lobster 100: RI

• Shell-diseased and healthy lobsters from the same sites

• Healthy lobsters from a reference site (ME) – lobster 100 study

• Approximately 185 samples analyzed– Hepatopancreas,

hemolymph, other tissues

What We Measured:

• Twenty five different trace elements, including:– Contaminant metals

Arsenic, cadmium, copper, chromium, nickel, lead, mercury

– Nutritive trace elementsCalcium, potassium, magnesium

– Other elementsAluminum, iron, manganese

What We Measured:

• Polychlorinated biphenyls (PCBs)– Selected congeners – 19

congeners

• Legacy organochlorine compounds– DDTs, aldrin, dieldrin,

endosulfan

• Polybrominated diphenyl ethers (PBDEs)

• Selected alkylphenols and bisphenol A See Poster

What We Have Learned:Contaminant Metals

• Regional Collections: RI, LIS, ME• Concentrations are generally similar between

diseased and non-diseased lobsters– Outliers occurred in all groups – Hepatopancreas and hemolymph

No Difference Between Diseased & Non-Diseased(RI Hepatopancreas)

Copper

SD NSD

Con

cent

ratio

n (

g/g

dry

wt.)

0

1000

2000

3000

4000

5000

Cadmium

SD NSD

0

20

40

60

80

100

120

Chromium

SD NSD

0

5

10

15

20

25Nickel

SD NSD

0

10

20

30

40

What We Have Learned:Lobster 100

• Two metals: Cr, Hg– Diseased > Non- Diseased and Reference– Two-ten fold higher in diseased

Lobster 100 Metal Concentrations Similar to Other Impacted Sites

Cd(g/g)

Cr(g/g)

Cu(g/g)

Ni(g/g)

Pb(g/g)

SD(31)

2.8 - 100(4.3)

0.08 – 35(4.23)

200 – 4,300(182)

0.32 – 31(1.28)

0.04 – 3.3(0.24)

NSD(30)

2.6 – 23(4.23)

0.08 – 1.2(0.41)

300 – 3,700(150)

0.27 – 5.1(0.96)

0.04 – 0.72(0.27)

Ref(19)

0.40 – 75(6.23)

0.05 – 1.3(0.28)

15 – 800(24)

0.79 – 3.6(1.39)

0.02 – 0.11(0.13)

Other Studies

Boston Harbor

1.2 – 6.6 0.1 – 2.5 37 - 640 0.51 – 0.85

LIS 5.4 – 50.8 0 – 0.75 180 – 3,880 0.90 – 5.97 1.2 – 15

Outfall (GOC)

11.5 + 7.5 0.9 + 0.5 380 + 500 4.4 + 0.7 350 + 77

Maine 1.8 - 59.8 0.07 - 1.3 12 - 500 0.07 - 2.4 0.02 - 0.46

What We Have Learned:Lobster 100

• Lobster sex (M/F) did not appear to be a significant factor in contaminant metal concentrations

• Sediments from the lobster 100 site had metal concentrations approaching levels of concern

As Cd Cr Cu Ni Pb

g/g) g/) g/g) g/) g/g) g/g)

Mean (n = 5) 15.3 0.2 104 31 420 106Std. dev. 1.3 0.1 2.2 2.8 25 82ERL1 8.2 1.2 81 34 30 47ERM2 70 9.6 370 270 50 2181ERL = effects range low = the l10th percentile of the concentration range where biological effects were observed in standardized sediment exposures2ERM = effects range median = the 50th percentile concentration range where biological effects were observed.

Sediment metals from the lobster 100 site approach and exceed levels of concern

What We Have Learned:Metals Partitioning

Hepatopancreas: As, Cd, Cu, Fe

Muscle:

Gills: Cr

Shell: Mn, Ni, Pb

Ovary:

Hemolymph:

Hepatopancreas metal concentrations sometimes exceed FDA action levels

Metals Action Level1 (ppm)

Hepatopancreas Concentration (ppm)

Muscle Concentration (ppm)

Arsenic 76 13 (0.75 - 170) 10 (6.9 – 14)

Cadmium 3 5.8 (0.08 -51) 0.91 (0 – 8.8)

Chromium 12 0.32 (0.03 – 17) 0.01 (0.01 – 0.02)

Lead 1.5 0.05 ( 0.01 – 0.11) 0.06 (0.01 – 0.57)

Mercury 1.0(methyl)

0.17 (0.04 – 0.49)(total)

0.07 (0.001 – 0.16)(total)

Nickel 70 1 0.82 (0.08 – 15) 1.0 (0.02 – 9.8)1 FDA Action Levels for Crustacea

Organic Contaminants

• Developed multi-residue method– Employed various separation and cleanup

technologies

• Hepatopancreas – Very lipid rich– Difficult to adequately clean up extract – Preliminary Data [See Poster]

Hexane: Acetonitrile

AluminaColumn

Gel Permeation Chromatography

Silica gelColumn

PCBs/OCs OCsAlkylphenolics

C-18 Column

PSA (primarySecondary amine)Column

Bisphenol ASteroidal Estrogens

Multi-residue Method

See Poster

Take Home Messages

• No strong relationship between metal body burden & disease state– Regionally, no differences b/w SD & NSD– Lobster 100 (RI), 2 metals SD> NSD>Ref

• Metal concentrations in hepatopancreas are similar to those of other impacted sites

• Metals in hepatopancreas (tomalley) sometimes exceed FDA action limits