1. Environmental Contaminants in Shell Diseased vs
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, ME
Department of Molecular and Cell Biology, University of Connecticut, Storrs, CT

2. 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

3. 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.)

4. Two Approaches LeBlanc and Prince: Laufer et al.
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

5. Our Hypothesis:
There is a relationship between epizootic shell disease and contaminant body burdens

6. 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)

7. Project Objectives Why Hepatopancreas? Why Hemolymph?
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

8. 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

9. What We Measured: Twenty five different trace elements, including:
Contaminant metals
Arsenic, cadmium, copper, chromium, nickel, lead, mercury
Nutritive trace elements
Calcium, potassium, magnesium
Other elements
Aluminum, iron, manganese

10. 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

11. 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

12. No Difference Between Diseased & Non-Diseased
(RI Hepatopancreas) 12

13. What We Have Learned: Lobster 100
Two metals: Cr, Hg
Diseased > Non- Diseased and Reference
Two-ten fold higher in diseased

14. Lobster 100 Metal Concentrations Similar to Other Impacted SitesCd
(mg/g) Cr Cu Ni Pb SD
(31)
(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
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
0.51 – 0.85
LIS
5.4 – 50.8
0 – 0.75
180 – 3,880
0.90 – 5.97
1.2 – 15
Outfall (GOC)
Maine

15. 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

16. Sediment metals from the lobster 100 site approach and exceed levels of concernAs Cd Cr Cu Ni Pb
(mg/g)
(mg/)
Mean
(n = 5)
15.3
0.2
104 31
420
106
Std. dev.
1.3
0.1
2.2
2.8 25 82
ERL1
8.2
1.2 81 34 30 47
ERM2 70
9.6
370
270 50
218
1ERL = effects range low = the l10th percentile of the concentration range where biological effects were observed in standardized sediment exposures
2ERM = effects range median = the 50th percentile concentration range where biological effects were observed. 16

17. What We Have Learned: Metals Partitioning

18. Hepatopancreas: As, Cd, Cu, Fe
Muscle:
Gills: Cr
Shell: Mn, Ni, Pb
Ovary:
Hemolymph:
Similar partitioning patterns observed between lobsters with and without shell disease
Hepatopancreas generally contained the greatest concentration of metal contaminants
Exoskeleton had highest tissue burden of Cr, Fe, Mn, Ni, Pb
Body burden of some contaminants reduced through molting

19. Hepatopancreas metal concentrations sometimes exceed FDA action levels
Metals
Action Level1 (ppm)
Hepatopancreas
Concentration (ppm)
Muscle Concentration (ppm)
Arsenic 76
13 ( )
10 (6.9 – 14)
Cadmium 3
5.8 ( )
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)
Nickel
70 1
0.82 (0.08 – 15)
1.0 (0.02 – 9.8)
1 FDA Action Levels for Crustacea

20. Organic Contaminants Developed multi-residue method Hepatopancreas
Employed various separation and cleanup technologies
Hepatopancreas
Very lipid rich
Difficult to adequately clean up extract
Preliminary Data [See Poster]

21. Multi-residue Method Hexane: Acetonitrile Alumina Column
Gel Permeation Chromatography
Silica gel
PCBs/OCs
OCs
Alkylphenolics
C-18 Column
PSA (primary
Secondary amine)
Bisphenol A
Steroidal Estrogens
Multi-residue Method
See Poster

22. 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