Transcript lobster shell disease

Is There a Link Between Epizootic Lobster Shell Disease and Contaminants?
Lawrence A. LeBlanc1, L. Brian Perkins, and Deanna Prince1
1School
THE PROBLEM
of Marine Sciences, 2Department of Food Science and Human Nutrition, 3Lobster Institute, University of Maine, Orono, Maine, USA
OBJECTIVES

Determine whether there is co-occurrence
between contaminant concentrations in
hepatopancreas/hemolymph and lobster shell
disease
The Lobster Institute
HYPOTHESES
•There
is a relationship between epizootic shell
disease and contaminant body burdens
Organic Contaminant Analysis
•Hemolymph


Lobsters with severely compromised exoskeletons
are being found from New York to Nova Scotia with
the greatest incidence in Southern New England
Is contaminant exposure correlated to the presence
of 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”
Why Hepatopancreas?
Concentrations of many trace elements are higher
than in muscle tissue
Elevated concentrations of lipophilic organic
contaminants
Why Hemolymph?
CANADA
Easier matrix to work with
Less labor-intensive sample preparation
Canada
can be used as a rapid screening
tool for contaminant body burdens
•Solvent extraction of tissue
•Accelerated solvent extraction
•Acetonitrile extracts analytes while leaving
behind lipids
•Partition against hexane
•Removal of lipids
•Alumina column
•Gel Permeation Chromatography
•Primary secondary amine (PSA)
•Separation of compound classes
•Solvent:Solvent partitioning:
Acetonitrile:Hexane
•Silica Gel Chromatography
•Instrumental Analysis
•Gas chromatography/mass spectrometry
GC/ECD, GC/MS
•Ion specific monitoring
•High performance liquid chromatography
•HPLC, LC-MS
Multiresidue analysis
Hexane:
Acetonitrile
Alumina
Column
C-18 Column
PSA (primary
Secondary amine)
Column
Silica gel
Column
Bisphenol A
Steroidal Estrogens
PCBs/OCs
OCs
Alkylphenolics
APPROACH
Gulf of Maine
Atlantic Ocean
Area of Interest
BACKGROUND
Alkylphenolics
•Laufer et al. (2005) postulate that alkylphenolic compounds
may be a causative agent in the onset of lobster shell
disease:
•High concentrations of alkylphenols detected in
sediments, lobster hemolymph and other tissues
(Biggers and Laufer (2004).
•Alkylphenolic compounds have endocrine-disrupting
properties, including interference with juvenile hormone
in invertebrates.
Contaminant Trace Metal Effects
•e.g., As, Cd, Cu, Cr, Hg, Ni, Pb, Se, Zn
•Lethal to invertebrates at high concentrations
•Sublethal effects at lower doses
•Affect immune & endocrine functions (Correa et al., 2005)
•Provoke oxidative stress (Valko et al., 2005)
Organic Contaminant Effects
•Several classes of organic compounds are known to interfere
with hormonal regulation
•PCBs, OCs (legacy pesticides)
•PBDEs
•Alkylphenols
•Estrogens (synthetic and natural)
•Lethal to invertebrates at high concentrations
•Sublethal effects at lower doses
•Interference with reproduction, thyroid functions
Lobster Collection & Tissue
Sampling
•Lobsters collected throughout the northeast
•Sea Grant
•State Agencies
•Industry contacts
•Matched collections of shell-diseased and healthy
lobsters
•Approximately 185 samples analyzed
CHEMICAL ANALYSES
Trace Metal Analysis
•Acid digestion of tissue:
•Microwave Acceleration
Reaction System (MARS)
Instrumental Analysis:
Metals:multi-elemental
analyses
ICP-AES: inductively
coupled plasma atomic
emission
ICP-MS: 10-100 x greater
sensitivity
•Mercury
•Direct mercury analyzer
Table 1. Hepatopancreas concentrations of different classes of
organic contaminants from lobster 100 samples
resulting from the multiresidue method
Shell Disease
Non- Disease
Reference
(n = 3)
(n = 3)
(n = 3)
ng/g
ng/g
ng/g
SPCB 19
1071 + 192
1040 + 570
86.3 + 22.5
p,p'-DDE
91.7 + 17.7
99.4 + 33.6
41.5 + 18.0
n-octylphenol
171 + 22.7
164 + 48
56.7 + 21.1
n-nonylphenol
429 + 26.6
247 + 65.1
30.0 + 18.0
ND
ND
ND
9.38, 7.51
7.77, 7.76
0.69, 4.59
1450-3500
MWRA
270-400
Ferarra (2005)
SPBDEs
BPA (mg/g)
SPCBs (Boston Hbr)
The effects of combined stressors
(e.g. metals + organics) are unknown
Nonylphenol (Adriatic Sea)