ARTICLE IN PRESS

The Science of the Total Environment xx (2003) xxx–xxx

Geographic variation in polychorinated biphenyl and

organochlorine pesticide concentrations in the blubber of
bottlenose dolphins from the US Atlantic coast
Larry J. Hansena, Lori H. Schwackeb,*, Greg B. Mitchumb, Aleta A. Hohnc,
Randall S. Wellsd, Eric S. Zolmanb, Patricia A. Fairb
United States Fish and Wildlife Service, 4001 North Wilson Way, Stockton, CA 95205, USA
a

b
National Oceanic and Atmospheric Administration, National Ocean Service, 219 Fort Johnson Road, Charleston,
SC 29412-9110, USA
c
National Oceanic and Atmospheric Administration, National Marine Fisheries Service, Southeast Fisheries Science Center,
101 Pivers Island Road, Beaufort, NC 28516, USA
d
Chicago Zoological Society, cyo Mote Marine Laboratory, Sarasota, FL 34236, USA

Accepted 23 May 2003

Abstract

Concentrations of polychorinated biphenyls (PCBs) and other organochlorine contaminants (OCs) were measured
in blubber collected from live bottlenose dolphins (Tursiops truncatus) at three sites along the United States Atlantic
coast. Dolphins were sampled via surgical biopsy during capture–release studies near Charleston, South Carolina and
Beaufort, North Carolina. Additional animals were sampled using remote biopsy techniques in estuarine waters near
Charleston and from the Indian River Lagoon, Florida. Overall concentrations of major contaminant groups were
found to vary between sites and mean concentrations of most OCs from male dolphins in the Indian River Lagoon
were less than half of those measured from Charleston and Beaufort males. Geometric mean total PCB concentrations
were 30, 27 and 14 mgyg lipid for male dolphins sampled in Beaufort, Charleston and the Indian River Lagoon,
respectively. Significant variation related to sex- and age-class, as well as geographic sampling location, was seen in
the PCB congener profiles. The measured PCB concentrations, although lower than those reported for stranded
animals from the 1987y1988 epizootic along the United States mid-Atlantic coast, are sufficiently high to warrant
concern for the health of dolphins from the sampled populations, particularly the animals near Charleston and
Beaufort.
䊚 2003 Elsevier Science B.V. All rights reserved.

Keywords: Bottlenose dolphins; Cetaceans; Organochlorines; Polychorinated biphenyls (PCBs); Marine mammals

*Corresponding author. Tel.: q1-843-762-8541; fax: q1-843-762-8700.

E-mail address: Lori.Schwacke@noaa.gov (L.H. Schwacke).

0048-9697/03/$ - see front matter 䊚 2003 Elsevier Science B.V. All rights reserved.
doi:10.1016/S0048-9697(03)00371-1
 ARTICLE IN PRESS
2 L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx
1. Introduction
An epizootic involving bottlenose dolphins (Tur-
siops truncatus) along the United States mid-
Atlantic coast in 1987–1988 and a finding of high
levels of organochlorine contaminants (OCs) in
the tissues of the recovered carcasses (Kuehl et
al., 1991) raised concerns regarding the potential
impact of contaminants on the health of Atlantic
coastal bottlenose dolphins. Morbillivirus-induced
disease was implicated as the primary cause of the
1987y1988 mass mortality event (Lipscomb et al.,
1994) and although a direct causal link between
OCs and the epizootic was not established, con-
taminant-induced immunosuppression was hypoth-
esized as a contributing factor (Wang et al., 1994).
Some OCs have been clearly demonstrated as
immunotoxicants in experimental studies (for
review see Ross, 2002) and given the high levels
measured from the stranded dolphins recovered
during this event, the role of OCs in facilitating
the emergence and spread of disease could have
been significant. Problematic is the fact that the
high contaminant levels were measured from
stranded animals that were recovered during the
event, therefore, the temporal sequence of exposure
and effect could not be established. It is impossible
to ascertain whether high OC burdens preceded
the epizootic or if OC concentrations in the dis-
eased and dying animals simply increased above
usual levels. Mobilization of lipid stores due to
nutritive stress and decreased metabolic capacity
linked to the animals’ diseased states could have
produced abnormally high OC concentrations in
their tissues (Aguilar et al., 1999).
Subsequent studies of contaminant concentra-
tions in other species of small cetaceans in the
western North Atlantic have been conducted along
the extreme northern US and Canadian coasts
using mass-stranded or by-caught animals (Muir
et al., 1988; Westgate et al., 1997; Weisbrod et al.,
2001). For bottlenose dolphins, one study has
investigated contaminant levels in stranded car-
casses along the coast of Florida (Watanabe et al.,
2000). However, because this analysis was per-
formed on stranded animals, the measured concen-
trations may not be indicative of OC
concentrations of live bottlenose dolphins from the
mid-Atlantic coastal stocks. Because disease is
often the cause of death in stranded animals, and
diseased animals may carry abnormal pollutant
loads, contaminant concentrations measured from
the tissues of stranded animals may not be accurate
indicators of the general population’s exposure
(Aguilar et al., 1999). Also, depending upon the
swimming patterns of the stranded animal prior to
death, and winds and currents after death, carcasses
may or may not have originated near the stranding
site. Additional issues arise from sampling a
decomposing carcass. Tissue decomposition and
subjection of the carcass to sun and wind exposure
may also alter pollutant concentrations. OC con-
centrations were found to decrease significantly in
the blubber of a dolphin carcass subjected to
natural environmental conditions for only a few
days (Borrell and Aguilar, 1990).
Blubber biopsy samples collected from live,
free-ranging cetaceans offer an alternative for
assessing OC concentrations and can provide sam-
ples that are representative of the target population.
Sampling of live animals can be performed by
remote dart biopsy or through capture–release
studies, which provide an additional opportunity
for health diagnostics and age determination. OC
concentrations measured from live animals permit
inferences to be made with regard to the distribu-
tion of contaminant concentrations in the general
population, thus allowing for statistical tests of
geographic and temporal trends. Knowledge of the
distribution of contaminant concentrations in the
general population also provides the necessary
basis for risk analyses to determine the magnitude
and probabilities of adverse health effects. In fact,
earlier capture–release efforts in the coastal waters
of Florida, Texas and North Carolina provided
measures of total PCB burdens, which were used
to estimate risks of reproductive failure in female
bottlenose dolphins (Schwacke et al., 2002).
This study uses OC concentrations obtained
from live, free-ranging bottlenose dolphins to
establish baseline values for a suite of PCB con-
geners and organochlorine pesticides and to deter-
mine differences in OC concentrations among three
distinct geographic sites along the US Atlantic
coast (Fig. 1). Blubber biopsies from bottlenose
dolphins, which were sampled as part of capture–
 ARTICLE IN PRESS
L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx 3

release efforts near Beaufort, North Carolina (NC)

and Charleston, South Carolina (SC), as well as
remote biopsies from dolphins near Charleston and
in the Indian River Lagoon, Florida (FL) were
incorporated into the analysis. Using multivariate
statistical techniques, we examine geographic var-
iation in the measured OC concentrations and
explore sources of variation in PCB congener
profiles.

2. Methods

2.1. Collection of samples

Capture–release studies were conducted in estu-

arine and coastal waters near Beaufort, NC in July,
1995, November, 1999 and April, 2000 and in
estuarine waters near Charleston, SC in October,
1999. These efforts were part of a program aimed
at assessing the health and defining stock structure
of coastal bottlenose dolphins along the southeast
US coast (Hohn, 1997). Methods for capture–
release followed those described by Scott et al.
(1990). Briefly, the animals were captured by
encircling them with a net in shallow waters and
restraining them by hand. Surgical biopsies meas-
uring approximately 2.5=1.5 centimeters (cm)
and weighing between 0.5–1.5 grams (g) were
taken from the left side of each animal. An attempt
was made to obtain a full blubber depth, so depth
of the biopsy varied between individuals, but was
generally between 1.0 and 2.0 cm. The biopsy site
was approximately 10 cm behind and 10 cm below Fig. 1. Map showing sampling locations of bottlenose dolphins
the posterior aspect of the dorsal fin. The site was along the US Atlantic Coast.
prepared with an antiseptic scrub and a local
anesthetic was injected in an ‘L’ block configura- affixed with a video camera to record the reaction
tion, approximately 4.0 cm ventral and anterior to of the animal and to collect images of dorsal fins
the intended biopsy site. A sterile scalpel and for individual identifications. The rifle fired a bolt
forceps were used to cut and extract the sample. which consisted of a thin-walled, hollow arrow
The biopsy samples were stored in pre-cleaned shaft and a hollow cylindrical collecting head
Teflon containers and frozen at y80 8C until approximately 2.5 cm in length and 1.0 cm in
chemical analyses could be performed. A tooth diameter. The sampling location on the body of
was extracted from each animal for age determi- the dolphin varied to some degree between indi-
nation (Hohn et al., 1989). viduals, but all the samples were collected within
Blubber samples from bottlenose dolphins in the an area that measured approximately 50 cm=30
Indian River Lagoon, FL were obtained via dart cm centered below the dorsal fin above the lateral
biopsy in January, 2002. Projectile biopsy samples midline. Prior to each use, the collecting head was
were obtained using a modified 0.22-caliber rifle scrubbed, soaked in a 10% bleach solution, then
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4 L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx

Table 1 s. The sample extract was brought to a volume of

Location, date and number of samples collected 100 ml. The percent lipid was then determined
Location Date Number of Sampling Total gravimetrically by drying a 5.0 ml aliquot. The
samples method remaining sample was reduced in volume to 0.5
ml. Lipids were removed from the persistent organ-
Male Female
ic contaminants by gel permeation chromatography
IRL, FL Jan 2002 9 2 Projectile 11 wtwo 300=21.20 mm tandem columns (Pheno-
Charleston, SC Oct 1999 5 6 Surgical menex, Torrance, CA) packed with phenogel, elut-
May 2000 2 Projectile ed with methylene chloride at a rate of 8 mlyminx.
May 2001 1 Projectile The sample was again reduced to 0.5 ml. Interfer-
Jul 2001 1 Projectile
Total 6 9 15
ing polar compounds were removed by Florisil
(US Silica Company, Berkeley Springs, WV) chro-
Beaufort, NC Jul 1995 17 14 Surgical matography w1 g, 6 ml Florisil solid phase extrac-
Nov 1999 4 2 Surgical
Apr 2000 5 5 Surgical tion cartridge (Argonaut Technologies, Foster City,
Total 26 21 47 CA), preconditioned with 5 ml of methylene chlo-
ride and eluted with 5 ml of 20% ethyl ethery80%
petroleum etherx. The sample was reduced to
rinsed twice, first with distilled water and then approximately 100 ml and placed in a GC vial for
with ethanol. Each sample was a cylindrical- analysis. The GC column was a DB-5 capillary
shaped core of skin and blubber, weighing approx- column (0.25 mm film thickness, 0.25 mm ID and
imately 0.5–1.0 g. Samples weighing less than 0.5 30 m in length, J and W ScientificyAgilent). The
g were excluded from the analysis. The sample Hewlett Packard 5972 mass spectrometer detector
was removed from the sampling tip with sterile was operated in the selected ion monitoring mode
forceps and scalpel. The same tools were used to and the samples were identified by retention time,
separate the skin from the blubber. The skin was target ion, and conformation ion ratios as compared
stored in a saturated salt solution of 20% dimethyl to known standards. The analytes were quantified
sulfoxide for determination of gender. The blubber against a 5-point linear curve. Standard reference
was stored in a pre-cleaned Teflon container and material (SRM 1945 Organics in Whale Blubber,
frozen at y808 C until chemical analyses could NIST) was analyzed for each group of eight
be performed. Along with the samples from the samples and compared with reference values for
Indian River Lagoon, four additional samples from quality assurance. The laboratory participated in
estuarine waters of Charleston, SC were obtained the Intercomparison Exercise for Persistent Organ-
using the remote biopsy method. The dates and ochlorine Contaminants in Marine Mammal Blub-
number of samples collected, along with the sam- ber sponsored by the National Institute of
pling method employed, are summarized in Table Standards and Technology to ensure quality control
1. and inter-laboratory comparison capability.
For samples obtained from the earliest live-
2.2. Extraction and gas chromatographymass capture (Beaufort, 1995), 15 PCB congeners were
spectrometry (GCyMS) analysis determined: (International Union for Pure and
Applied Chemistry wIUPACx 噛’s 18, 28, 52, 66,
Blubber samples were macerated with sodium 101y90, 105, 118, 128, 138y163y164, 153, 156,
sulfate and added to a 33 ml Dionex accelerated 180, 187, 206, 209). For subsequent analyses,
solvent extraction (ASE) cell (Dionex, Salt Lake methods were expanded to determine an additional
City, UT). An internal standard mix was added to 13 congeners: (IUPAC 44, 49, 87, 95, 99, 149,
the ASE cell, and the cell was sealed for extraction. 151, 169, 170y190, 183, 194, 195, 201). The DDT
The sample was extracted at 100 8C and 2000 psi compounds (2,49-DDD, 4,49-DDD, 2,49-DDE, 4,49-
and flushed with a 50% volume of methylene DDE, 2,49-DDT, 4,49-DDT) and chlordane-related
chloride for four cycles and finally purged for 80 compounds (heptachlor epoxide, cis-chlordane,
 ARTICLE IN PRESS
L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx
trans-chlordane and trans-nonachlor) were quan-
tified, as well as the chlorinated pesticides aldrin,
dieldrin, alpha- and beta-hexachlorocyclohexane
and hexachlorobenzene. With the exception of the
1995 Beaufort samples, cis-nonachlor, methoxy-
chlor, mirex, endosulfan and endosulfan sulfate
were also quantified.
2.3. Sex and age determination
For all the animals from the two live-capture
sites, sex was determined from direct observation
of the genital region of the animals and age was
estimated from counts of the growth layers in teeth
as described by Hohn et al. (1989). For the remote
(projectile) biopsy samples, sex determination
using DNA from skin was performed using meth-
ods described by Rosel (2003). No age estimates
were possible from the remote biopsy sampling.
2.4. Statistical analyses
Statistical analyses were performed using Statis-
tica (Statsoft, 1999). Contaminant concentrations
on a wet weight basis were divided by the fraction
of lipid in the sample to provide a lipid-normalized
concentration. A value of one half of the detection
limit was assigned for samples that measured
below the limits of detection. Summary variables
were defined for PCBs, DDT and chlordane com-
pounds. For all comparative analysis between geo-
graphic regions, 8PCBs was defined as the sum
of the 15 common PCB congeners. The number
of PCB congeners included in the summation was
limited to these 15 so that the early samples
obtained from Beaufort, NC in 1995 could be
included in the comparative analyses. 8DDT was
defined as the sum of the six DDT group com-
pounds and 8chlordane was defined as the sum
of heptachlor epoxide, cis-chlordane, trans-chlor-
dane and trans-nonachlor. The concentration of
cis-nonachlor was not included in the summation
of chlordane compounds because it was not meas-
ured in the 1995 Beaufort, NC samples.
Although we did not quantify the highly toxic
non-ortho PCBs, toxic equivalents to 2,3,7,8-
TCDD (TEQs) were estimated based on the mono-
ortho PCBs, IUPAC 噛’s 105, 118 and 156. We
5
used toxic equivalency factors (TEFs) of 0.0001,
0.001 and 0.0005 for PCB 105, 118 and 156,
respectively (Van den Berg et al., 1998).
Numerous studies have shown a sharp decrease
in OC burdens for females when they reach repro-
ductive maturity (reviewed by Aguilar et al.,
1999), occurring at approximately 8–10 years of
age (Wells and Scott, 1999). The observed
decrease in OC burdens in reproductively active
females is related to the offloading of contaminants
to their young through parturition and more nota-
bly lactation (Aguilar et al., 1999). Because of the
disparity in OC burdens for sexually mature and
immature females, it is important to stratify female
samples by reproductive status. Unfortunately,
reproductive histories were not known for the
majority of the sampled animals. For this reason,
samples with known ages were stratified into broad
age-classes, with animals less than 10 years being
classified as juveniles (both sexes combined) and
animals 10 years or older classified as adult males
or adult females. Since ages could not be deter-
mined for animals that were sampled via projectile
biopsy, these samples were classified as unknown-
age males and unknown-age females.
Prior to performing geographic analyses, differ-
ences in the major contaminant groups (8PCBs,
8DDT, 8chlordane and dieldrin) were established
between sex- and age- classes using a multivariate
analysis-of-variance (MANOVA) technique. Vari-
ables were log-transformed to meet assumptions
of normality. Because ages were known only for
live-captured animals, this portion of the analysis
included only samples from the Charleston and
Beaufort capture–release studies. Geographic sam-
pling location was also included as a co-factor.
Because of unequal sample sizes, the GT-2 method
(Hochberg, 1974) was used for pairwise compari-
son of the sexyage classes (i.e. adult male vs.
adult female, adult male vs. juvenile and juvenile
vs. adult female). Proportions of each major con-
taminant group were also examined graphically to
determine differences between sexyage classes.
2.4.1. Geographic variation in major contaminant
groups
To examine differences between geographic
regions in the absolute concentrations of major
 ARTICLE IN PRESS
6 L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx
contaminant groups, a MANOVA was conducted
using 8PCBs, 8DDT, 8chlordane and dieldrin.
Because of the strong influence of reproductive
status on OC concentrations in females, and the
fact that the ages of remote-biopsied females could
not be determined, only males were included in
this portion of the analysis. Variables were log-
transformed to meet assumptions of normality. The
MANOVA was followed with univariate F-tests
for each contaminant group and a pairwise com-
parison of means between the three sites using the
GT-2 method (Hochberg, 1974).
2.4.2. Sources of variation in PCB congener
profiles
To investigate sources of variation in PCB con-
gener profiles, two separate analyses were con-
ducted. The first, aimed at establishing differences
in congener profiles between sex and age classes,
used regression analysis of age vs. concentrations
of PCB structural groups. Previous studies have
classified PCB congeners into structural groups
based on substitution patterns (Tanabe et al., 1988;
Boon et al., 1992, 1994) with an underlying
assumption that congeners within the same struc-
tural group exhibit similar metabolic degradation.
PCB group I congeners have G2 ortho-chlorines
and no vicinal hydrogen atoms (IUPAC 153, 180,
187, 206, 209), while PCB group II congeners
have vicinal hydrogen atoms in the ortho and meta
positions in combination with 2–3 ortho-chlorines
(IUPAC 128, 138). Studies have shown that ceta-
ceans exhibit little or no metabolic capability for
congeners belonging to these two structural groups
(Tanabe et al., 1988; Boon et al., 1994). PCB
group III congeners have ortho or meta vicinal
hydrogen atoms in combination with 0–1 ortho-
chlorines and are purportedly metabolized by
CYP1A isozymes (IUPAC 28, 66, 105, 118, 156),
while PCB group IV congeners have vicinal hydro-
gens in the meta and para positions in combination
with two ortho-chlorines and are believed to be
metabolized by CYP2B isozymes (IUPAC 18, 52,
101) (Tanabe et al., 1988; Boon et al., 1994).
These groupings were used to examine relation-
ships between age and PCB burdens. A regression
analysis of age against concentration of each PCB
group was performed, using sampling location
(Beaufort or Charleston) as a binary covariate.
Analysis for males and females was performed
separately.
Secondly, a principal components analysis
(PCA) was conducted to determine structure in
PCB congener profiles and to examine the identi-
fied structure with regard to sampling location.
The proportions of each congener in relation to
the sum of all PCB congeners were used as
variables for the PCA. The 1995 samples from
Beaufort were not included in the analysis so that
the larger suite of PCB congeners could be utilized.
Congeners for which greater than 60% of the
samples measured concentrations below the meth-
od detection limit were excluded from the analysis.
Because of noted differences in concentrations and
accumulation patterns for adult females related to
reproductive state, only males and juveniles were
included in the PCA. The extracted components
from the PCA were examined graphically to look
for differences related to geographic location and
age.
3. Results
3.1. Organochlorine concentrations
PCBs, DDT, chlordane and dieldrin were the
predominant compounds in all three sites (Tables
2–4). In order of decreasing concentration, dol-
phins from all three sites had PCBsyDDT)chlor-
dane)dieldrin, with concentrations of SPCBs and
SDDT being generally very similar.
As anticipated, concentrations of all contami-
nants varied between sexyage classes (P-0.0001)
(Fig. 2). Concentrations measured from adult
females were significantly lower than those meas-
ured from both juveniles (P-0.001 for all contam-
inant groups) and adult males (P-0.001 for all
contaminant groups). Concentrations of SPCBs,
SDDT and Schlordane in adult males were slightly
higher than those measured in juveniles. The dif-
ferences were not statistically significant (SPCBs
Ps0.13, SDDT Ps0.13, Schlordane Ps0.10),
but the lack of significance could be attributable
to the limited sample size for adult males. Dieldrin
concentrations between adult males and juveniles
were very similar (Ps0.97).
Table 2
Geometric mean, minimum and maximum of organochlorine concentrations (ngyg lipid) in blubber biopsies from live-captured bottlenose dolphins sampled near
Beaufort, North Carolina. Juveniles represent both males and females less than 10 years old. Adult males and adult females are 10 years or older

Analyte Beaufort
Juveniles Adult Males Adult Females
a a
Valid N Mean Min Max Valid N Mean Min Max Valid N Meana Min Max
Lipid (%) 29 28.3 14.0 57.2 5 29.5 19.0 37.7 6 30.0 16.0 54.0

L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx
PCB 18 29 35.9 7.17 168 5 21.3 7.17 168 6 8.14 2.78 16.0
PCB 28 29 2439 8.57 441 5 23.0 8.57 441 6 26.9 13.9 51.9
PCB 44 8 20.9 9.18 65.0 3 14.7 9.18 65.0 2 20.6 12.9 33.0
PCB 49 8 59.9 11.8 245 3 19.3 11.8 245 2 28.1 12.8 61.7
PCB 52 29 1139 319 2348 5 1442 319 2348 6 108 13.3 730
PCB 66 29 140 16.3 406 5 94.5 16.3 406 6 61.2 14.4 183
PCB 87 8 34.9 10.6 117 3 18.2 10.6 117 2 31.0 11.6 82.8

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PCB 95 8 733 357 1472 3 1141 357 1472 2 101 11.9 851
PCB 99 8 2273 1335 4460 3 4265 1335 4460 2 625 125 3127
PCB 101 29 961 208 2495 5 535 208 1495 6 170 62.6 735
PCB 105 29 323 101 604 5 299 101 604 6 55.6 15.6 583
PCB 118 29 1280 640 2560 5 8987 640 2560 6 226 62.5 2028
PCB 128 29 761 168 3067 5 1108 468 3067 6 89.8 40.0 1006
PCB 138 29 3731 388 26341 5 6683 388 26341 6 480 101 2563
PCB 149 8 2236 1372 4432 3 3992 1372 4432 2 724 175 2986
PCB 151 8 875 545 1651 3 1767 545 1651 2 205 40.4 1031
PCB 153 29 8315 2115 38985 5 15732 2115 38985 6 1295 496 11268
PCB 156 29 72.0 15.1 143 5 76.9 15.1 143 6 42.3 13.7 125
PCB 170 8 239 18.9 812 3 1240 18.9 812 2 226 76.1 670
PCB 180 29 3642 1050 18770 5 7201 1050 18770 6 813 328 3606
PCB 183 8 505 190 1248 3 2455 190 1248 2 359 101 1270
PCB 187 29 3750 1468 16911 5 6980 1468 16911 6 768 304 3915
PCB 194 8 371 212 793 3 790 212 793 2 341 144 806
PCB 195 8 138 10.2 364 3 255 10.2 364 2 113 62.8 202
PCB 201 8 209 7.82 588 3 470 7.82 588 2 169 89.4 318
PCB 206 29 613 212 1743 5 795 212 1743 6 540 244 1161
PCB 209 29 193 5.03 517 5 369 5.03 517 6 449 158 1338
sum 15 PCBs 29 27231 11281 110193 5 44152 11281 110193 6 5644 2469 29158
sum 27 PCBs 8 28275 15910 52248 3 53276 15910 52248 2 11631 3332 40596
2,49-DDD 29 209 18.2 520 5 197 18.2 520 6 44.8 18.8 87.5
2,49-DDE 29 277 10.1 660 5 388 10.1 660 6 43.5 9.11 219
2,49-DDT 29 216 20.0 7292 5 624 20.0 7292 6 303 28.2 1639
4,49-DDD 29 3114 1048 5950 5 2987 1048 5950 6 381 125 1842
4,49-DDE 29 23487 9952 80252 5 46338 9952 80252 6 2726 1163 15042
4,49-DDT 29 429 120 1319 5 415 120 1319 6 ND (128)

7
 8
Table 2 (Continued)
Analyte Beaufort
Juveniles Adult Males Adult Females
a a
Valid N Mean Min Max Valid N Mean Min Max Valid N Meana Min Max
sum DDT 29 28849 11311 87281 5 51906 11311 87281 6 3988 1592 18957
cis-chlordane 29 345 14.8 3548 5 158 14.8 3548 6 61.3 33.5 210
heptachlor epoxide 29 182 7.54 730 5 222 7.54 730 6 47.0 9.73 172

L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx
trans-nonachlor 29 3069 1398 8925 5 6406 1398 8925 6 269 160 1882
trans-chlordane 29 49.5 29.3 85.7 5 56.3 29.3 85.7 6 ND (24)
sum chlordane 29 3908 1619 10320 5 7022 1619 10320 6 548 262 2321
cis-nonachlor 8 319 10.1 849 3 534 10.1 849 2 159 52.0 485
methoxychlor 8 346 36.4 8420 3 42.8 36.4 8420 2 ND (31)
mirex 8 310 14.5 1244 3 508 14.5 1244 2 161 148 174
dieldrin 29 968 86 2343 5 991 86 2343 6 139 52.0 530
ND (77.0) ND (77.0) ND (77.0)

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aldrin 29 5 6
alpha HCH 29 ND (40.0) 5 ND (40.0) 6 ND (40.0)
beta HCH 29 ND (38.9) 5 ND (38.9) 6 ND (38.9)
hexachlorobenzene 29 57.1 15.0 129 5 ND (36.0) 6 ND (36.0)
endosulfan 8 ND (39.8) 3 ND (39.8) 2 ND (39.8)
endosulfan sulfate 8 ND (31.9) 3 ND (31.9) 2 ND (31.9)
a
Geometric means (except for lipid %)
Table 3
Geometric mean, minimum and maximum of organochlorine concentrations (ngyg lipid) in blubber biopsies from live-captured and dart-biopsied bottlenose dolphins
sampled near Charleston, South Carolina. Juveniles represent both males and females less than 10 years old. Adult males and adult females are 10 years or older

Analyte Charleston
Juveniles Adult Males Adult Females
a a
Valid N Mean Min Max Valid N Mean Min Max Valid N Meana Min Max
Lipid (%) 2 31.0 25.0 38.0 4 19.9 9.7 33.7 5 29.7 18.6 47.5

L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx
PCB 18 2 18.9 10.7 33.2 4 27.0 12.2 123 5 19.6 8.63 128
PCB 28 2 48.8 46.6 51.2 4 34.9 16.8 132 5 23.5 11.9 88.7
PCB 44 2 40.5 21.3 77.3 4 32.8 23.0 54.0 5 17.6 11.1 28.2
PCB 49 2 62.3 21.1 184 4 66.2 53.5 81.7 5 41.1 23.2 90.4
PCB 52 2 871 384 1976 4 1304 706 2047 5 100 11.4 938
PCB 66 2 136 80.6 230 4 100 60.2 139 5 27.8 12.3 121
PCB 87 2 48.4 19.0 123 4 50.9 43.0 62.3 5 23.2 9.9 59.1

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PCB 95 2 947 428 2093 4 1409 703 2366 5 113 21.5 945
PCB 99 2 2342 961 5709 4 4065 1881 6934 5 447 111 2662
PCB 101 2 595 554 640 4 6785 544 887 5 194 68.2 654
PCB 105 2 85.5 31.2 235 4 675 215 308 5 36.3 16.2 173
PCB 118 2 1030 883 1202 4 1193 974 1667 5 307 100 1180
PCB 128 2 760 266 2170 4 1296 573 2016 5 110 57.9 268
PCB 138 2 660 629 692 4 1490 204 4547 5 122 39.9 688
PCB 149 2 2467 1131 5385 4 3888 1769 6831 5 559 169 2688
PCB 151 2 936 381 2300 4 1652 715 2809 5 142 23.9 1096
PCB 153 2 7355 2898 18664 4 14144 6172 24074 5 1893 560 9706
PCB 156 2 119 79.4 177 4 28.0 16.5 57.1 5 18.7 11.7 29.8
PCB 170 2 515 190 1397 4 1044 445 1646 5 61.7 24.8 255
PCB 180 2 2357 885 6275 4 5670 2409 10010 5 938 337 4375
PCB 183 2 926 321 2668 4 2026 858 3117 5 139 50.8 467
PCB 187 2 2598 1055 6397 4 5586 2424 9650 5 939 326 4154
PCB 194 2 269 100 725 4 930 368 1420 5 213 69.1 658
PCB 195 2 134 60.8 296 4 307 142 543 5 88.0 34.5 257
PCB 201 2 203 81.5 506 4 553 239 936 5 91.2 15.1 415
PCB 206 2 328 167 644 4 1032 472 1698 5 478 246 696
PCB 209 2 107 59.0 195 4 359 152 628 5 233 166 380
sum 15 PCBs 2 18135 8385 39223 4 34285 16347 57873 5 5856 2026 22144
sum 27 PCBs 2 27385 12423 60363 4 50379 23642 84632 5 7981 2708 31190
2,49-DDD 2 202 126 323 4 132 33.6 270 5 35.8 16.1 147
2,49-DDE 2 232 119 453 4 339 191 532 5 39.7 7.79 235
2,49-DDT 2 96.2 29.9 310 4 114 50.2 262 5 90.1 24.1 2956
4,49-DDD 2 2459 1358 4453 4 2910 2095 4702 5 405 46.8 2180
4,49-DDE 2 14344 6057 33968 4 29123 14955 48148 5 3412 914 16397
4,49-DDT 2 346 179 668 4 ND (137) 5 ND (137)

9
 10
Table 3 (Continued)
Analyte Charleston
Juveniles Adult Males Adult Females
a a
Valid N Mean Min Max Valid N Mean Min Max Valid N Meana Min Max
sum DDT 2 17780 7869 40176 4 33102 17691 54474 5 4445 1153 22167
cis-chlordane 2 390 298 510 4 307 237 433 5 61.7 28.5 207
heptachlor epoxide 2 48.8 16.0 149 4 96.0 17.3 445 5 20.0 8.32 128

L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx
trans-nonachlor 2 2599 888 7611 4 5919 2798 9866 5 537 115 3518
trans-chlordane 2 ND (41.4) 4 ND (41.4) 5 ND (41.4)
sum chlordane 2 3379 1389 8221 4 6606 3387 10553 5 744 195 3816
cis-nonachlor 2 689 352 1348 4 812 561 1183 5 125 40.8 537
methoxychlor 2 ND (30.6) 4 ND (30.6) 5 83.1 32.2 618
mirex 2 233 88.0 615 4 663 306 1101 5 166 83.8 592
dieldrin 2 924 661 1291 4 772 582 1173 5 177 65.2 537
ND (45.9) ND (45.9) ND (45.9)

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aldrin 2 4 5
alpha HCH 2 ND (8.90) 4 ND (8.90) 5 ND (8.90)
beta HCH 2 ND (59.4) 4 ND (59.4) 5 ND (59.4)
hexachlorobenzene 2 51.7 34.8 76.8 4 ND (17.2) 5 ND (17.2)
endosulfan 2 ND (39.8) 4 ND (39.8) 5 ND (39.8)
endosulfan sulfate 2 ND (31.9) 4 ND (31.9) 5 ND (31.9)
a
Geometric means (except for lipid %)
 ARTICLE IN PRESS
L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx 11

Table 4
Geometric mean, minimum and maximum of organochlorine concentrations (ngyg lipid) in blubber biopsies from dart-biopsied
bottlenose dolphins sampled in the Indian River Lagoon, Florida

Analyte IRL
Males Females
a
Valid N Mean Min Max Valid N Meana Min Max
Lipid (%) 9 39.4 30.1 46.0 2 46.8 40.6 54.0
PCB 18 9 11.7 10.0 15.3 2 9.8 8.52 11.3
PCB 28 9 47.1 12.9 110 2 26.4 10.5 66.5
PCB 44 9 15.2 11.4 40.7 2 11.2 9.7 12.9
PCB 49 9 78.7 39.2 508 2 160 53.9 476
PCB 52 9 594 478 743 2 257 91.7 722
PCB 66 9 41.8 13.4 140 2 98.1 58.4 165
PCB 87 9 35.0 10.8 61.5 2 16.7 8.70 32.0
PCB 95 9 445 323 577 2 168 65.0 431
PCB 99 9 1794 1442 2566 2 575 236 1402
PCB 101 9 141 11.5 385 2 196 87.4 441
PCB 105 9 156 86.0 270 2 131 57.9 298
PCB 118 9 518 253 976 2 412 168 1011
PCB 128 9 445 342 683 2 152 67.7 341
PCB 138 9 788 78.5 1650 2 419 198 889
PCB 149 9 513 35.3 1368 2 395 186 841
PCB 151 9 488 389 751 2 176 81.5 380
PCB 153 9 6631 5022 9832 2 2408 1209 4796
PCB 156 9 18.2 12.6 77.4 2 11.9 10.3 13.7
PCB 170 9 24.0 14.7 232 2 56.9 16.6 194
PCB 180 9 549 38.5 2854 2 772 502 1185
PCB 183 9 710 457 1172 2 303 195 470
PCB 187 9 2690 1943 4180 2 1158 709 1889
PCB 194 9 540 393 945 2 299 246 363
PCB 195 9 95.3 63.9 131 2 18.8 6.76 52.5
PCB 201 9 318 222 489 2 26.7 5.19 138
PCB 206 9 704 420 1189 2 524 411 667
PCB 209 9 104 71.2 157 2 15.6 3.33 72.9
sum 15 PCBs 9 14443 10203 21639 2 6807 3726 12436
sum 27 PCBs 9 20038 14724 27948 2 9264 5041 17024
2,49-DDD 9 19.4 16.6 25.4 2 31.0 18.8 51.1
2,49-DDE 9 65.4 47.8 118 2 ND (7.40) 6.85 9.11
2,49-DDT 9 60.7 26.1 767 2 146 28.2 754
4,49-DDD 9 451 251 683 2 202 54.8 743
4,49-DDE 9 11598 6630 23141 2 3627 1515 8685
4,49-DDT 9 ND (137) 2 ND (137) 127 169
sum DDT 9 12647 8294 23894 2 4313 1794 10366
cis-chlordane 9 92.4 58.2 138 2 16.8 9.8 28.8
heptachlor epoxide 9 17.6 9.00 166 2 ND (7.90) 7.31 9.73
trans-nonachlor 9 2698 1867 4484 2 860 392 1889
trans-chlordane 9 ND (41.4) 2 ND (41.4) 38.3 51.0
sum chlordane 9 2902 2079 4619 2 967 481 1944
cis-nonachlor 9 332 217 465 2 182 73.6 448
methoxychlor 9 67.6 33.3 5304 2 ND (30.6) 28.3 37.7
mirex 9 61.0 11.9 168 2 20.3 9.9 42.6
dieldrin 9 222 86.5 355 2 ND (61.9) 57.3 76.2
 ARTICLE IN PRESS
12 L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx

Table 4 (Continued)
Analyte IRL
Males Females
a
Valid N Mean Min Max Valid N Meana Min Max
aldrin 9 ND (45.9) 2 ND (45.9) 42.5 56.5
alpha HCH 9 ND (8.90) 2 ND (8.90) 8.24 11.0
beta HCH 9 ND (59.4) 2 ND (59.4) 55.0 73.2
hexachlorobenzene 9 ND (17.2) 2 ND (17.2) 15.9 21.2
endosulfan 9 ND (39.8) 2 ND (39.8)
endosulfan sulfate 9 ND (31.9) 2 ND (31.9)
a
Geometric means (except for lipid %)

SDDT accounted for the greatest proportion of
the measured OCs in juveniles and adult males,
while the greatest proportion in adult females came
from the PCB compounds (Fig. 3).
Concentrations of methoxychlor, an insecticide
used for agriculture as well as for home gardening
and pets, were below detection limits for the
majority of samples. However, a few dolphins
sampled from Beaufort had extremely high
methoxychlor concentrations. Samples from seven
of the Beaufort dolphins measured methoxychlor
concentrations greater than 0.1 mgyg lipid, three
of which were greater than 1.0 mgyg lipid and one
of which was nearly 10 mgyg lipid.
3.2. Differences in major contaminant groups
related to sampling location
The MANOVA revealed a highly significant
effect of sampling location on contaminant con-
centrations (P-0.0001). Follow-up univariate
ANOVAs yielded significant differences between
sampling locations for SPCBs (Ps0.001), SDDT
(Ps0.0003) and dieldrin (P-0.0001), but no

Fig. 2. Geometric mean and 95% confidence interval for contaminant concentrations measured in blubber of dolphins sampled in
Charleston and Beaufort (combined). Juvenile group represents combined males and females 10 years of age or less. Letters (a–h)
represent statistically equivalent (PG0.05) groups as determined by pairwise comparisons of each contaminant group between
sexyage classes.
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L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx 13

Fig. 3. Mean proportion of each major contaminant group measured from juveniles (age-10 years), adult males and adult females.
Samples from Charleston and Beaufort were combined. Whiskers represent standard errors.

difference in Schlordane concentrations was found
between sites (Ps0.22). The results of pairwise
comparisons indicated that the IRL dolphins had
lower concentrations of SPCBs, SDDT and dield-
rin as compared to the Beaufort dolphins (PF0.01
for all three contaminants) (Fig. 4). Differences
between the IRL and Charleston dolphins with
regard to concentrations of SPCBs and SDDT
were only marginally significant (Ps0.05 and Ps
0.06, respectively), likely due to the limited sample
sizes. Charleston dolphins did have much higher
dieldrin concentrations as compared to the IRL

Fig. 4. Geometric means and 95% comparison limits for contaminant concentrations measured in blubber of male dolphins from
the three sampling sites. Letters (a–g) represent statistically equivalent (PG0.05) groups as determined by pairwise comparisons
for each contaminant group between sites.
 ARTICLE IN PRESS
14 L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx
dolphins (Ps0.002). None of the contaminant
concentrations were significantly different between
Beaufort and Charleston dolphins (P)0.1 for all
three contaminants).
The sample of males from the Beaufort site
contained a disproportionate number of young
animals (77% of sampled males were 10 years or
less in age) and there was some concern that this
could cause a negative bias in the mean OC
concentrations for Beaufort males. Since the com-
parison between the Beaufort and IRL dolphins
showed Beaufort animals to have significantly
higher OC concentrations, the potential negative
bias obviously did not significantly influence this
portion of the analysis. However, the null finding
for the comparison of Beaufort and Charleston
concentrations could be erroneous. Only 20% of
the sampled males from Charleston were 10 years
of age or less, compared with 77% of the males
from Beaufort. Because additional information
regarding age was available for the Beaufort and
Charleston males, a follow-up multivariate analysis
of covariance (MANCOVA) was performed, add-
ing age as a continuous covariate. In this analysis,
controlling for age, the difference in overall con-
taminant concentrations between Beaufort and
Charleston males was significant (Ps0.003). Uni-
variate ANOVAs indicated that the concentrations
of SPCBs and SDDT were significantly higher in
Beaufort males (SPCBs Ps0.02, SDDT P-0.01)
as compared to Charleston males, but concentra-
tions of dieldrin and Schlordane did not differ
between the two sites (Ps0.35 and Ps0.16,
respectively).
Mean total TEQ values were 215 (range 84–
339), 172 (range 131–226) and 86 (range 41–
142) pgyg lipid for males from Beaufort,
Charleston and the IRL, respectively. Mean total
TEQ values were lower for females: 148 (range
17–307), 89 (range 18–212) and 83 (range 29–
136) pgyg lipid for Beaufort, Charleston and the
IRL, respectively.
3.2.1. Sources of variation in PCB congener
profiles
The concentration of each individual congener
as a proportion of the total PCB concentration was
graphed for comparison between age- and sex-
classes and sampling locations (Fig. 5). Propor-
tional PCB profiles varied considerably between
age- and sex-classes, particularly between adult
females vs. adult males or juveniles. Adult females
showed an increased proportion of octa-, nona-
and deca-biphenyls and a lesser proportion of PCB
153 as compared to adult males and juveniles.
3.2.2. Concentration of PCB groups vs. age
Polynomial functions were fit to the group III
concentrations separately for males and females
(Fig. 6a). Including population as a binary covar-
iate in the regression tested the effect of differing
sampling locations (Beaufort vs. Charleston), but
this parameter was not found to contribute signif-
icantly to the model (males Ps0.31, females Ps
0.64). Parameters for age and age2 were significant
for both males and females (all cases, P-0.01)
and the overall models were significant for both
males (Ps0.004) and females (P-0.0001).
Similarly, for group IV PCBs (Fig. 6b), the
effect of sampling location was not significant, but
parameters for both age and age2 were significant
for both males and females (all cases, P-0.01).
The overall model for females was highly signifi-
cant (P-0.0001), but due to the high degree of
variability in the males’ concentrations, the model
fit was not as good (Ps0.02).
For group I and II concentrations, a polynomial
function (Fig. 6c) provided an acceptable fit to
the data for females (P-0.0001), and both para-
meters for age and age2 contributed significantly
to the model (P-0.0001 for both). However, a
model for the concentration of group I and II PCBs
in males was difficult to determine due to the high
degree of variability in the measured concentra-
tions. A polynomial function did not provide an
acceptable fit to the data (Ps0.10). A linear
regression model provided an adequate fit for the
data (Ps0.002), however, a large proportion of
the variation was left unexplained (R 2s0.33) and
the model residuals were unequally distributed,
with much greater variance for younger animals.
Concentrations in males under the age of 10–12
did not exhibit a clear relationship with age.
Around the age of 12–15, concentrations did
appear to rise, but the number of data points in
this age range was insufficient to provide a stable
 ARTICLE IN PRESS
L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx 15

Fig. 5. Mean proportion of each PCB congener measured from bottlenose dolphins sampled near Beaufort, Charleston and the Indian
River Lagoon. Bars represent mean proportion of total PCBs accounted for by each congener. Whiskers represent minimum and
maximum values.
 ARTICLE IN PRESS
16 L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx
Fig. 6. Age vs. concentration of summed congeners of various PCB groups. Hollow squares represent females and filled circles
represent males.
model fit. Due to these statistical issues, we did
not provide a fitted function for group I and II
concentrations in males.
3.2.3. Principal components analysis for PCB
congener profiles
The PCA extracted four components that cumu-
latively accounted for 78% of the total variance.
The first principal component, PC1, accounted for
33% of the variance, while PC2 accounted for
23%. Scores for the first two components were
graphed in order to compare scores between the
three populations (Fig. 7). PC1 completely sepa-
rated the IRL dolphins from the Beaufort and
Charleston dolphins, with all IRL samples having
positive scores, and all Beaufort and Charleston
dolphins having negative scores. PC1 had the
highest positive loadings for the heptachlorobi-
phenyl 187, the octachlorobiphenyls 194 and 201,
and the nonachlorobiphenyl 206 (Fig. 8a), and the
highest negative loading for the hexachlorobiphen-
yls 149 and 151.
Differences in PC2 scores were not readily
explained by sampling location, but did appear to
be related to age. For samples for which age could
be determined, negative PC2 scores were from
dolphins under the age of 7 years, while all of the
samples with positive PC2 scores were from dol-
phins 9 years of age or greater. Given the results
of the PCB group regressions against age, it was
anticipated that differences between adult and
juvenile males would be a source of variation in
the analysis. The most significant loadings for PC2
were PCB 183, a group I congener, and PCB 138
and 128, both group II congeners (Fig. 8b). The
loadings for these three congeners were all posi-
tive, therefore, a higher score for PC2 would
indicate a higher proportion of these congeners.
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L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx
Fig. 7. Scores for first two principal components. Only males were included in the principal components analysis. Samples within
the oval are all from dolphins less than 7 years.
The scores for PC2 appeared to increase with age,
consistent with the hypothesis of increasing con-
centrations of the non-metabolizable group I and
II congeners with age.
4. Discussion
Consistent with numerous prior studies, our data
showed the lowest concentrations of OCs in
females of reproductive age. Adult males and
juveniles were found to have the highest concen-
trations, with values measured from adult males
being slightly higher than those from juveniles.
Differences in summary OC concentrations were
not statistically significant between adult males
and juveniles, but this would likely change if
larger sample sizes were analyzed.
Proportions of each major contaminant group
varied between sexyage classes, with a higher
proportion of SDDT seen in juveniles and adult
males, and a higher proportion of PCBs seen in
adult females (Fig. 3). Prior studies have also
shown higher ratios of SDDTySPCBs in adult
males as compared to adult females (summarized
by Aguilar et al. (1999)). Selective partitioning
during reproductive transfer has been suggested by
several studies of pinnipeds (Addison and Brodie,
1987; Beckmen et al., 1999) and is believed to be
due to the differing physical and chemical prop-
17
erties of the various OC compounds. An early
study of pinnipeds suggested that DDT groups are
more efficiently transferred from blubber to milk
as compared to the PCBs (Addison and Brodie,
1987). A later study by Salata et al. (1995) found
a higher proportion of maternal SDDT as com-
pared to SPCBs in an unborn fetus collected from
a single female bottlenose dolphin that stranded
with a full term calf. Our data also provide
evidence for selective partitioning during repro-
ductive transfer.
4.1. Geographic variation
Overall concentrations of OCs were similar
between dolphins sampled near Beaufort and
Charleston, although the Beaufort concentrations
were slightly higher once age of the animals was
considered. The lowest OC concentrations were
measured from dolphins within the IRL. In fact,
mean concentrations measured from IRL males
were less than half of those measured from both
Charleston and Beaufort males for all major OC
groups except for the chlordane compounds.
There were variables, such as sampling season
and sampling method, which could be considered
as confounding factors between sites. For example,
all of the samples from the IRL dolphins were
taken during the winter (January), while samples
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18 L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx

Fig. 8. Factor loadings for first two principal components. ‘*’ indicates loadings greater than 0.70.

from Charleston and Beaufort were taken during
spring, summer and fall months. Unfortunately, the
number of samples from each site and each sexy
age class were insufficient to thoroughly test the
effect of season, but for juveniles sampled from
the Beaufort site, OC concentrations were lower
in animals sampled in the early spring (April), as
compared to those sampled in both July and
November (data not shown). Lower concentrations
of OCs might be expected in the winter and early
spring months, because at this time of the year,
the dolphins have increased their insulating blub-
ber layer, thereby possibly decreasing the concen-
tration of contaminants. In the Beaufort juveniles,
lipid concentrations were higher in samples taken
in April (43% lipid) and November (41% lipid)
as compared to those taken in July (25% lipid). It
is unlikely that the degree of difference in OC
concentrations seen between the Beaufort and the
IRL dolphins could be attributable to seasonal
 ARTICLE IN PRESS
L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx
variation, but the confounding effect of season
should be quantified for future studies.
The possibility of sampling method as a con-
founding factor should also be considered. Careful
interpretation of OC concentrations measured via
remote biopsies has been suggested due to concern
that lipid amount and composition may be strati-
fied vertically within the cetacean blubber layer
(Koopman et al., 1996; Tilbury et al., 1997; Hobbs
et al., 2003). Vertical stratification of lipid types
has been noted in the blubber of cetaceans (Koop-
man et al., 1996) and other marine mammals (Best
et al., 2003). If OC concentrations and profiles
vary along with the lipid composition and only the
superficial blubber layer is sampled using remote
biopsy, then a bias could be introduced. Tilbury et
al. (1997) found higher OC concentrations in the
innermost subsample as compared to the middle
and outermost subsamples of blubber from two
stranded harbor porpoises. However, these results
were not consistent for a third harbor porpoise in
the same study, which showed no substantial dif-
ferences among OC concentrations measured in
inner, middle and outer blubber subsamples. A
study using the carcasses of three stranded bottle-
nose dolphins from Charleston, South Carolina
also divided blubber into three layers and exam-
ined differences in OC concentrations and lipid
composition between the layers. In contrast to the
study of harbor porpoise, the study of bottlenose
dolphins found higher OC concentrations in the
outermost blubber layer, although the differences
were slight and varied between individuals (Fair,
P., unpublished data). Results from studies of St.
Lawrence beluga whales have also been inconsis-
tent, with some whales showing lower concentra-
tions in the outermost blubber and others showing
the reverse (reviewed by Hobbs et al., 2003).
Unfortunately, all of the aforementioned studies
have been conducted using stranded animals and
therefore, may not be indicative of variation in the
blubber layers of live cetaceans. In view of the
conflicting information and the uncertainty with
regard to the validity of generalizations from
stranded to live animals, it would be preferable to
obtain core samples that are a full blubber depth
to accurately represent OC concentration in blub-
ber. Alternatively, relative measures for geographic
19
comparisons involving full depth and projectile
biopsy samples might be obtained by subsampling
the outer portion of the full depth sample to match
the projectile biopsies.
In this study, the surgical biopsies obtained from
live-captured dolphins were full blubber depth
samples, although the same could not be confirmed
for most of the remote-biopsy samples. Smaller
remote-biopsy samples (less than 0.5 g weight)
were excluded from the analysis so that all blubber
samples ranged from 0.5 g to just over 1.0 g,
regardless of the sampling method. Although hav-
ing a larger amount of blubber does not necessarily
guarantee a full blubber depth, it increases the
likelihood and would exclude samples of only the
outermost blubber layer that might be obtained
from a remote-biopsy if the dart were to penetrate
at an angle, thus giving only a ‘glancing’ sample.
For the Charleston site, both remote and surgical
biopsy samples were obtained. Although the num-
ber of remote biopsy samples (4) was insufficient
for a rigorous statistical analysis of sampling meth-
od differences, some cursory comparisons can be
made. OC concentrations measured in the few
remote-biopsy samples from dolphins in the
Charleston estuaries were comparable to concen-
trations measured from surgical biopsies obtained
from captured dolphins in the same area and were
much higher than remote biopsies taken from
dolphins in the IRL. Specifically, the total PCB
concentration measured from the single remote-
biopsied male in Charleston was 48.3 mgyg lipid.
This is in complete concordance with the median
total PCB concentration for live-captured males
(all ages combined) from Charleston, which was
53.0 mgyg lipid (quartiless23.6–61.0 mgyg lip-
id), but is nearly twice the maximum concentration
measured from remote-biopsied males in the IRL
(28 mgyg lipid). Comparison of the samples from
females is more tenuous because there is disparity
between mature and immature females, and age or
reproductive status could not be definitively deter-
mined for the remote-biopsied females. However,
one of the remote-biopsied females was observed
with a calf, estimated to be 1.5–3 years of age
based on its size. The total PCB concentration
measured from this female’s blubber (3.96 mgyg
lipid) was very similar to concentrations measured
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20 L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx

Fig. 9. Comparison of PCB blubber concentrations from this study with previous study of bottlenose dolphins obtained after the
1987y1988 unusual mortality event (UME) (Kuehl et al., 1991). Filled squares and hollow triangles represent median and whiskers
represent minimum and maximum values. Values represent samples from males and juveniles only. For comparability between
studies, PCBs are represented by the sum of the five individual PCB congeners as reported by Kuehl et al. (1991).

from surgical biopsy samples taken from other
Charleston females that were captured with calves
within the same age range (2.71–5.60 mgyg lipid).
Concentrations measured from the other two
remote-biopsied females (17.49 and 32.40 mgyg
lipid) were within the range of those measured
from immature females from the Charleston live-
capture group (15.52–60.36 mgyg lipid). These
two females were not in the company of a calf at
the time that they were sampled, although this
does not necessarily imply that they were not
reproductively mature. In any case, these two
samples from remotely-biopsied females in
Charleston were higher than even the maximum
concentration measured from remotely-biopsied
females in the IRL. The fact that these few remote
biopsy samples from Charleston were comparable
with other surgical biopsies from Charleston yet
higher than the maximum values from IRL remote
biopsies, suggest that the concentration differences
between sites were likely attributable to location
rather than sampling method.
Other studies have suggested that remote biopsy
sampling may provide an inaccurate profile of OCs
present (Hobbs et al., 2003). However, results of
our PCA indicate no substantial difference in the
congener profile for the remote-biopsied male as
compared to the other males from Charleston,
which were sampled via surgical biopsy. In the
graph of scores for the first two principal compo-
nents (Fig. 7), all of the Charleston samples,
including the remote-biopsy sample, were closely
aligned on both axes and very easily differentiated
from the remote-biopsy samples from the IRL. In
short, although the number of samples for com-
parison was limited, the concentrations measured
from remote-biopsied vs. the surgical-biopsied dol-
phins in Charleston provided no evidence to sug-
gest that the alternate biopsy methods introduced
bias into the measurement of OC concentrations.
Although the potential confounding of sampling
method cannot be ruled out in the comparison of
concentrations between the IRL (remote-biopsied)
dolphins vs. the Charleston and Beaufort (surgical-
biopsied) dolphins, the influence is likely minimal.
4.2. Local sources
There were seven dolphins sampled near Beau-
fort that were found to have unusually high
methoxychlor concentrations. Of these seven, three
have been sighted since the captures in northern
Pamlico Sound, north of Beaufort and their move-
ment patterns suggest that they may be residents
 ARTICLE IN PRESS
L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx
of the Sound (Rich Mallon-Day, Nags Head Dol-
phin Watch, pers. communication). The Environ-
mental Protection Agency has listed a site along
the adjacent Pamlico River on its National Priori-
ties List (NPL) based on pesticide contamination
in soil, sediment, surface water and groundwater.
One of the particular contaminants of concern for
the NPL site is methoxychlor. Further research,
looking at OC concentrations in prey species from
the Pamlico Sound and other nearby estuarine
systems, is needed to determine if there is a point
source of contamination linked to the high meth-
oxychlor concentrations from the blubber of these
few dolphins.
4.3. Sources of variation in OC proportions and
PCB congener profiles
Analysis of variation in contaminant patterns
has been suggested as a supplemental tool, to be
combined with genetic and morphological analy-
ses, for discriminating stocks of marine mammals
(reviewed by Aguilar, 1987). Prior studies have
demonstrated significant geographic variation in
organochlorine and trace element concentrations
from cetacean blubber and skin. A study of minke
whales in the southern hemisphere suggested the
usefulness of trace elements in skin to elucidate
population structure (Kunito et al., 2002). West-
gate et al. (1997) found regional differences in
PCB congener profiles from the blubber of harbour
porpoises, suggesting that contaminant concentra-
tions might be useful in discriminating stocks of
this species in the northwest Atlantic. Similarly,
our results suggest that OC patterns could be useful
in discriminating stocks of coastal bottlenose
dolphins.
Our results showed higher proportions of the
superhydrophobic octa-, nona- and deca-biphenyls
and a lower proportion of the recalcitrant congener
PCB 153 in adult females as compared to adult
males and juveniles (Fig. 5). Congener 153 is not
readily metabolized by marine mammals but has
been shown to be a dominant component of their
milk (Pomeroy et al., 1996; Beckmen et al., 1999).
The elevated ratios of the superhydrophobic con-
geners measured in adult females could be due to
the offloading of PCB 153, but not of the octa-,
21
nona- and deca-biphenyls, through lactation. Dif-
fering reproductive states of females would explain
the high degree of variability in these ratios
between individuals. Prior studies have suggested
that the food chain transfer of superhydrophobic
congeners is less effective due to steric hindrance
and membrane permeation resistance (Kannan et
al., 1998; Maruya and Lee, 1998). The same
principles would likely apply for transfer of these
superhydrophobic compounds from blubber to
milk, making them more resistant to reproductive
transfer. Our observed pattern in proportional PCB
congener profiles for adult females provides evi-
dence to support this type of selective partitioning.
The PCA, which included only males, showed
a distinct separation of IRL samples from the
Beaufort and Charleston samples related to PCBs
187, 194, 201 and 206. A high proportion of these
particular congeners is characteristic of Aroclor
1268, a highly chlorinated PCB mixture that has
been found in relatively few US sites (Kannan et
al., 1997). Other major PCB formulations contain
relatively minor amounts of these highly chlori-
nated congeners. Although the IRL males main-
tained a higher proportion of these particular
congeners, Charleston males actually had higher
absolute concentrations. Beaufort males had the
lowest concentrations of these congeners. This
could indicate that both IRL and Charleston ani-
mals have some source of exposure to Aroclor
1268, but that Charleston animals are exposed to
additional PCB sources that contribute to the lower
chlorinated congeners. The estuary where the
Charleston animals were sampled is more open to
the adjacent ocean environment and is under a
much greater tidal influence as compared to the
IRL. Therefore, dolphins residing in these Charles-
ton waters could be consuming prey items that
have been exposed to the lower chlorinated PCBs
transported via atmospheric or oceanic currents.
A Superfund site on the southeastern coast of
Georgia has been determined to be a major source
of Aroclor 1268 contamination (Kannan et al.,
1997). The Superfund site is approximately 180
miles southwest of Charleston and 280 miles
northwest of our IRL sampling site, following the
coastline. It is possible that Aroclor 1268 from
this site near Brunswick, Georgia contributed to
 ARTICLE IN PRESS
22 L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx
the PCB exposure profiles noted in the Charleston
and the IRL dolphins, although generally such
highly chlorinated biphenyls tend to be less mobile
and remain close to the source of the contamination
(Wania and Mackay, 1996). Further research and
additional samples are needed to examine potential
PCB sources for these coastal areas.
4.3.1. Concentration of PCB groups vs. age
By separating PCB congeners into structural
groups, we found a strong relationship between
age and tissue concentration for both males and
females. Our results for PCB groups III and IV
are consistent with a model predicting a decline in
PCB concentrations with age related to the increas-
ing mass of the animal. Hickie et al. (1999)
developed a physiologically based pharmacokinet-
ic (PBPK) model for total PCB concentrations in
marine mammals and applied their model to data
for beluga whales (Delphinapterus leucas) from
West Greenland and the St. Lawrence Estuary.
Their model suggested that from birth through the
nursing period, a sharp increase in PCB body
burden could be expected as a result of the high
doses of PCBs that calves receive via their moth-
ers’ lipid-rich milk. Weaning, which takes place
around the age of 1.5 years in beluga whales and
1.5–2 years in bottlenose dolphins, represents a
transition from the highly contaminated lipid-rich
milk to a fish diet containing lower concentrations
of lipophilic contaminants. The period of rapid
growth that follows, along with the lowered intake
of contaminants, purportedly results in a dilution
effect that works to decrease overall contaminant
burdens. The dilution effect diminishes as the
growth rate eventually declines toward zero, at
which time PCB concentrations are predicted to
rise again with age, depending on the degree of
contamination in prey. Females are an exception
to this pattern once the age of reproductive matur-
ity has been reached. Sharp declines in female
PCB burdens are predicted following the birth of
their first calf and the subsequent period of lacta-
tion. Levels are then predicted to remain low
throughout the female’s reproductive lifespan.
The minimum age of the sampled individuals in
our study was 1.5 years so patterns of accumula-
tion in newborns and calves could not be con-
firmed from our data. Our data did show group III
and IV congener concentrations decreasing from
the minimum sample age (1.5 years) up to approx-
imately 10–12 years of age in both males and
females (Fig. 6a,b). Growth models have shown
that it is approximately at this same age that
bottlenose dolphins reach asymptotic length indi-
cating the cessation of growth (Stolen et al., 2002).
Therefore, it is likely that this decrease in juvenile
concentrations is to some degree linked to a
dilution process resulting from the addition of
mass, as suggested by Hickie et al.’s PBPK model.
The dilution effect is likely enhanced by the
capacity, albeit limited, of individuals to metabo-
lize group III and IV PCBs via CYP1A and
CYP2B enzymes, respectively. Past the age of
growth cessation, concentrations in males began
to rise and eventually reached levels equivalent to
those seen in the very young animals. Because the
oldest male in our study was 25 years of age, it is
impossible to say whether the increasing trend
would continue or whether concentrations would
eventually reach saturation and begin to level off.
For females, group III and IV congener concentra-
tions dropped dramatically around the age of 10
at the time of first reproduction and then remained
at a plateau. One older female, age 49 years, had
a very high concentration of both group III and
IV PCBs, similar to levels seen in very young
animals. This would suggest that at some point,
concentrations in females also began to increase,
similar to bioaccumulation patterns observed in
the males. Such an increase could be related to an
extended time between births in older females,
however, such inferences from the current data,
which have no samples from animals between the
ages of 30–49, would be questionable.
The age-related patterns for the non-metaboliz-
able group I and II PCBs were not so clear (Fig.
6c). In females, a sharp decrease near the age of
first reproduction was noted, consistent with other
PCB groups and almost certainly related to the
offloading of burdens through lactation. For males,
concentrations appear to rise once growth ceases,
but it is unclear whether or not concentrations
undergo a decline prior to this age. It is probable
that the ‘dilution’ effect is not as strong for this
PCB group because it is based only on the increase
 ARTICLE IN PRESS
L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx
in mass and possibly a decrease in dose, but is not
supplemented by any elimination process.
4.4. Comparison with prior analyses and threshold
values
In order to compare PCB concentrations meas-
ured from this study with those measured from
stranded animals during and after the 1987y1988
mass mortality event, we calculated the sum of the
five PCB congeners (IUPAC 噛’s 118, 128, 138,
153 and 180), which were reported by Kuehl et
al. (1991). Median concentrations of the summed
five congeners measured from all three sites in
this study were lower than concentrations reported
from the stranded bottlenose dolphins collected
from the 1987y1988 epizootic (Fig. 9). Concentra-
tions of 4,49-DDE were fairly comparable between
the Beaufort, Charleston and the epizootic animals.
The OC concentrations reported by Kuehl et al.
(1991) were from bottlenose dolphin carcasses
collected from North Carolina, Virginia and Mar-
yland, so it is possible that some of the disparity
in PCB loads is due to geographic variation.
However, as previously discussed, OC loads meas-
ured from stranded animals suffer from a number
of issues, making comparisons over time or geo-
graphic location questionable.
Ross et al. (2000) suggest a toxic threshold
value of 255 pgyg lipid total TEQ based on an
immunotoxicological study of pinnipeds (Ross et
al., 1995). The mean total TEQ values from this
study were below this threshold, although the TEQ
values we report are an underestimate of the true
total TEQ because we did not quantify concentra-
tions of non-ortho PCBs. Non-ortho PCBs were
found to contribute minimally (less than 1% in
adult males) to the total TEQ s calculated for
bottlenose dolphins recovered from the 1987y1988
mortality event (Kuehl et al., 1994), so it is
unlikely that the magnitude of the underestimation
is large. Some individual dolphins (approx. 35%
of Beaufort males) did exceed the threshold TEQ
even though non-ortho PCBs were not included in
our TEQ calculation.
Alternatively, examining total PCBs, mean con-
centrations for animals sampled from both Beau-
fort and Charleston exceeded toxic threshold
23
values of 17 mgyg lipid weight proposed by
Kannan et al. (2000) and 14.8 mgyg proposed by
Schwacke et al. (2002). Fitting lognormal distri-
butions to total PCB loads measured from males
at each site as described by Schwacke et al.
(2002), we found that approximately 90% of males
from both the Charleston and Beaufort sites and
79% of males from the IRL exceeded the 17 mgy
g toxic threshold. This suggests that the popula-
tions sampled from this study are at high risk for
adverse health effects related to PCB exposure.
Furthermore, given that many of the organochlo-
rine pesticides, such as DDT and chlordane, have
been linked with deleterious health effects similar
to those associated with PCBs (e.g. reproductive
or immunological impairment, Lahvis et al., 1995),
there may be additive or even synergistic effects
to consider.
5. Conclusions
Using blubber biopsies, we have provided bas-
eline organochlorine measurements for live bottle-
nose dolphins from three sites along the US
Atlantic coast. The measured PCB concentrations,
although lower than those reported for stranded
animals from the 1987y1988 epizootic, are suffi-
ciently high to warrant concern for the health of
dolphins from the sampled populations, particular-
ly the animals near Charleston and Beaufort.
Because of the potential for downward bias in OC
concentrations measured from dolphins in the IRL
due to both sampling season (winter) and sampling
method (remote-biopsy), the possibility of under-
estimation of risk for IRL dolphins cannot be ruled
out. Additional sampling of dolphins in the IRL
to explore the influence of these confounding
variables is warranted.
We demonstrated differences in sexyage classes
that emphasize the importance of having informa-
tion on age, sex and reproductive status for the
interpretation of OC concentrations. Our results
indicate some differences in overall concentrations
of the major OC groups in dolphin blubber, as
well as substantial variation in PCB congener
profiles, related to geographic sampling location.
Although larger samples would be required, our
 ARTICLE IN PRESS
24 L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx
findings support the use of contaminant profiles as
a potential tool for stock discrimination.
Acknowledgments
The authors would like to thank the researchers
who participated in the capture–release efforts in
Charleston and Beaufort, particularly Larry Fulford
who safely captured the animals for sampling, and
Forrest Townsend and Jay Sweeney who per-
formed the surgical biopsy and tooth extraction
procedures. The DNA analysis to determine sex
for the remote-biopsied animals was performed by
Patricia Rosel and Sarah Kingston of the Marine
Mammal Molecular Genetics Lab at the National
Marine Fisheries Service (NMFS) Southeast Fish-
eries Science Center. We would like to express our
gratitude to Marilyn Mazzoil, Robin Friday, Rene
Varela, Steve McCulloch and Greg Bossart of
Harbor Branch Oceanographic Institute and to
Tony Martinez and Jenny Litz of the NMFS
Southeast Fisheries Science Center for their time
and effort in collecting remote biopsies in the IRL.
We thank Rich Mallon-Day for providing infor-
mation on sightings of freeze-branded animals in
Pamlico Sound via the Nags Head Dolphin Watch
website (www.dolphin-watch.com) and through
personal communications. We are also grateful to
John Kucklick for reviewing this manuscript and
for providing valuable insight for the interpretation
of our results. Samples were collected under Sci-
entific Research Permit 噛 960 issued to NMFS
Southeast Fisheries Science Center for the live-
captures and Scientific Research Permit 噛 779–
1339, also issued to NMFS Southeast Fisheries
Science Center, for the projectile biopsy sampling.
Funding was provided by the National Marine
Fisheries Service, the National Ocean Service and
the Chicago Zoological Society. The National
Ocean Service (NOS) does not approve, recom-
mend or endorse any proprietary product or mate-
rial mentioned in this publication. No reference
shall be made to NOS or to this publication
furnished by NOS in any advertising or sales
promotion which would indicate or imply that
NOS approves, recommends or endorses any pro-
prietary product or proprietary material mentioned
herein or which has as its purpose any intent to
cause directly or indirectly the advertised product
to be used or purchased because of NOS
publication.
References
Addison R, Brodie P. Transfer of organochlorine residues from
blubber through the circulatory system in milk in the
lactating grey seal, Halichoerus grypus. Can J Fish Aquat
Sci 1987;44:782 –786.
Aguilar A. Using organochlorine pollutants to discriminate
marine mammal populations: a review and critique of the
methods. Mar Mamm Sci 1987;3(3):242 –262.
Aguilar A, Borrell A, Pastor T. Biological factors affecting
variability of persistent pollutant levels in cetaceans. In:
Reijnders P, Aguilar A, Donavan G, editors. Chemical
pollutants and cetaceans, vol. special issue 1, Journal of
Cetacean Research and Management. Cambridge, UK; 1999.
pp. 83–116.
Beckmen K, Ylitalo G, Towell R, Krahn M, O’Hara T, Blake
J. Factors affecting organochlorine contaminant concentra-
tions in milk and blood of northern fur seal (Callorhinus
ursinus) dams and pups from St. George Island, Alaska. Sci
Total Environ 1999;231:183 –200.
Best N, Bradshaw C, Hindell M, Nichols P. Vertical stratifi-
cation of fatty acids in the blubber of southern elephant
seals (Mirounga leonina): implications for diet analysis.
Comp Biochem Physiol B 2003;134:253 –263.
Boon JP, Arnhem EV, Jansen S, Kannan N, Petrick G, Schulz
D, Duinker JC, Reijnders PJH, Goksoyr A. The toxicoki-
netics of PCBs in marine mammals with special reference
to possible interactions of individual congeners with the
cytochrome P450-dependent monooxygenase system: an
overview. In: Walker CH, Livingstone DR, editors. Persist-
ent pollutants in marine ecosystems. Tarrytown, NY, USA:
Pergamon Press, 1992. p. 119 –159.
Boon P, Oostingh I, Meer Jvd, Hillebrand MTJ. A model for
the bioaccumulation of chlorobiphenyl congeners in marine
mammals. Eur J Pharm 1994;270:237 –251.
Borrell A, Aguilar A. Loss of organochlorine compounds in
the tissues of a decomposing stranded dolphin. Bull Environ
Contam Toxicol 1990;45:46 –53.
Hickie B, Mackay D, Koning JD. Lifetime pharmacokinetic
model for hydrophobic contaminants in marine mammals.
Environ Toxicol Chem 1999;18(11):2622 –2633.
Hobbs K, Muir D, Michaud R, Beland P, Letcher R, Norstrom
R. PCBs and organochlorine pesticides in blubber biopsies
from free-ranging St. Lawrence River Estuary beluga whales
(Delphinapterus leucas), 1994–1998. Environ Pollut
2003;122:291 –302.
Hochberg Y. Some generalizations of the T-method in simul-
taneous inference. J Multivar Anal 1974;4:224 –234.
Hohn A, Scott M, Wells R, Sweeney J, Irvine A. Growth
layers in teeth from free-ranging, known-age bottlenose
dolphins. Mar Mamm Sci 1989;5(4):315 –342.
 ARTICLE IN PRESS
L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx
Hohn AA. Design for a multiple-method approach to determine
stock structure of bottlenose dolphins in the mid-Atlantic.
NOAA Technical Memorandum. NMFS-SEFSC-401, 1997,
p. 22.
Kannan K, Blankenship AL, Jones PD, Giesy JP. Toxicity
reference values for the toxic effects of polychlorinated
biphenyls to aquatic mammals. Hum Ecol Risk Assess
2000;6(1):181 –201.
Kannan K, Maruya K, Tanabe S. Distribution and characteri-
zation of polychlorinated biphenyl congeners in soil and
sediments from a Superfund site contaminated with Aroclor
1268. Environ Sci Technol 1997;31:1483 –1488.
Kannan K, Nakata H, Stafford R, Masson G, Tanabe S, Giesy
JP. Bioaccumulation and toxic potential of extremely hydro-
phobic polychlorinated biphenyl congeners in biota collected
at a Superfund site contaminated with Aroclor 1268. Environ
Sci Technol 1998;32:1214 –1221.
Koopman H, Iverson S, Gaskin D. Stratification and age-
related differences in blubber fatty acids of the male harbour
porpoise (Phocoena phocoena). J Comp Physiol
1996;165(8):628 –639.
Kuehl DW, Haebler R, Potter C. Chemical residues in dolphins
from the US. Atlantic coast including Atlantic bottlenose
obtained during the 1987y88 mass mortality. Chemosphere
1991;22(11):1071 –1084.
Kuehl DW, Haebler R, Potter C. Coplanar PCB and metal
residues in dolphins from the US Atlantic coast including
Atlantic bottlenose obtained during the 1987y88 mass mor-
tality. Chemosphere 1994;28(6):1245 –1253.
Kunito T, Watanabe I, Yasunaga G, Fujise Y, Tanabe S. Using
trace elements in skin to discriminate the populations of
minke whales in southern hemisphere. Mar Environ Res
2002;53:175 –197.
Lahvis G, Wells R, Kuehl D, Stewart J, Rhinehart H, Via C.
Decreased lymphocyte responses in free-ranging bottlenose
dolphins (Tursiops truncatus) are associated with increased
concentrations of PCBs and DDT in peripheral blood.
Environ Health Perspect 1995;103:67 –72.
Lipscomb T, Schulman F, Moffett D, Kennedy S. Morbilliviral
disease in an Atlantic bottlenose dolphin (Tursiops trunca-
tus) from the 1987–88 epizootic. J Wildl Dis 1994;30:567 –
571.
Maruya K, Lee R. Biota-sediment accumulation and trophic
transfer factors for extremely hydrophobic polychorinated
biphenyls. Environ Toxicol Chem 1998;17(12):2463 –2469.
Muir D, Wagemann R, Grift N, Norstrom R, Simon M, Lien
J. Organochlorine chemical and heavy metal contaminants
in white-beaked dolphins (Lagenorhynchus albirostris) and
pilot whales (Globicephala melaena) from the coast of
Newfoundland, Canada. Arch Environ Contam Toxicol
1988;17:613 –629.
Pomeroy P, Green N, Hall A, Walton M, Jones K, Harwood J.
Congener-specific exposure of grey seal (Halichoerus gry-
pus) pups to chlorinated biphenyls during lactation. Can J
Fish Aquat Sci 1996;53:1526 –1534.
Rosel P. PCR-based sex determination in Odontocete cetace-
ans. Conservation Genetics 2003 (accepted).
25
Ross P. The role of immunotoxic environmental contaminants
in facilitating the emergence of infectious diseases in marine
mammals. Hum Ecol Risk Assess 2002;8(2):277 –292.
Ross P, Swart RD, Reijnders P, Loveren HV, Vos J, Osterhaus
A. Contaminant-related suppression of delayed-type hyper-
sensitivity and antibody responses in harbor seals fed herring
from the Baltic Sea. Environ Health Perspect 1995;103:162 –
167.
Ross PS, Ellis GM, Ikonomou MG, Barrett-Lennard LG,
Addison RF. High PCB concentrations in free-ranging Pacif-
ic killer whales, Orcinus orca: effects of age, sex and dietary
preference. Mar Pollut Bull 2000;40(6):504 –515.
Salata GG, Wade TL, Sericano JL, Davis JW, Brooks JM.
Analysis of Gulf of Mexico bottlenose dolphins for organ-
ochlorine pesticides and PCBs. Environ Pollut 1995;88:167 –
175.
Schwacke LH, Voit EO, Hansen LJ, Wells RS, Mitchum GB,
Hohn AA, Fair PA. Probabilistic risk assessment of repro-
ductive effects of polychorinated biphenyls on bottlenose
dolphins (Tursiops truncatus) from the southeast United
States coast. Environ Toxicol Chem 2002;21(12):2752 –
2764.
Scott M, Wells R, Irvine A. A long-term study of bottlenose
dolphins on the west coast of Florida. In: Leatherwood S,
Reeves R, editors. The Bottlenose Dolphin. San Diego, CA,
USA: Academic Press, 1990. p. 235 –244.
Stolen M, Odell D, Barros N. Growth of bottlenose dolphins
(Tursiops truncatus) from the Indian River Lagoon System,
Florida, USA. Mar Mamm Sci 2002;18(2):348 –357.
Tanabe S, Watanabe S, Kan H, Tatsukawa R. Capacity and
mode of PCB metabolism in small cetaceans. Mar Mamm
Sci 1988;4(2):103 –124.
Tilbury K, Stein J, Meador J, Krone C, Chan S. Chemical
contaminants in harbor porpoise (Phocoena phocoena) from
the north Atlantic coast: tissue concentrations and intra- and
inter-organ distribution. Chemosphere 1997;34(9y10):2159 –
2181.
Van den Berg M, Birnbaum L, Bosveld ATC, Burnstrom B,
Cook P, Feeley M, Giesy JP, Hanberg A, Hasegawa R,
Kennedy SW, Kubiak T, Larsen JC, van Leeuwen FXR,
Liem AKD, Nolt C, Peterson RE, Poellinger L, Safe S,
Schrenk D, Tillitt D, Tysklind M, Younes M, Waern F,
Zacharewski T. Toxic equivalency factors (TEFs) for PCBs,
PCDDs, PCDFs for humans and wildlife. Environ Health
Perspect 1997;106(12):775 –791.
Wang KR, Payne PM, Thayer VG. Coastal stock(s) of Atlantic
bottlenose dolphin: status review and management. NOAA
Technical Memorandum. NMFS-OPR-4, 1994, p. 121.
Wania F, Mackay D. Tracking the distribution of persistent
organic pollutants. Environ Sci Technol 1996;30(9):A 390 –
A 396.
Watanabe M, Kannan K, Takahashi A, Loganathan BG, Odell
DK, Tanabe S, Giesy JP. Polychorinated biphenyls, organo-
chlorine pesticides, tris(4-chlorphenyl)methane, and tris(4-
chlorophenyl)methanol in livers of small cetaceans stranded
along Florida coastal waters, USA. Environ Toxicol Chem
2000;19(6):1566 –1574.
 ARTICLE IN PRESS
26 L.J. Hansen et al. / The Science of the Total Environment xx (2003) xxx–xxx
Weisbrod AV, Shea D, Moore MJ, Stegeman JJ. Species, tissue
and gender-related organochlorine bioaccumulation in white-
sided dolphins, pilot whales and their common prey in the
northwest Atlantic. Mar Environ Res 2001;51(1):29 –50.
Wells R, Scott M. Bottlenose dolphin Tursiops truncatus
(Montagu, 1821). In: Ridgeway S, Harrison R, editors.
Handbook of marine mammals, the second book of dolphins
and porpoises, vol. 6. San Diego, CA, USA: Academic
Press, 1999. p. 137 –182.
Westgate A, Muir D, Gaskin D, Kingsley M. Concentrations
and accumulation patterns of organochlorine contaminants
in the blubber of harbour porpoises, Phocoena phocoena,
from the coast of Newfoundland, the Gulf of St. Lawrence
and the Bay of FundyyGulf of Maine. Environ Pollut
1997;95(1):105 –119.