Challenges in small cetacean telemetry: an attempt at developing a remotely deployed attachment device for single-pin dorsal fin satellite transmitters.

Satellite telemetry is critical for collecting fine-scale temporal and spatial data on individual animals that has broad-scale applicability at population and species levels. There have been significant advances in the remote deployment of satellite telemetry devices on large cetacean species. However, the development of comparable remote attachment methodologies for small cetaceans is still limited. Currently, satellite tag attachment for small cetaceans requires manual capture that increases the risk to the target animal, can be logistically challenging, and cost prohibitive. The goal of this project was to develop a novel tool to remotely attach single-pin satellite telemetry devices to the dorsal fin of individual small cetaceans. Three different spring-loaded designs and one pneumatic version of the remote attachment device were built in an iterative process to identify a successful deployment methodology. Ultimately, as a result of logistical challenges associated with a Category 5 hurricane, the COVID-19 pandemic, and engineering complexities related to dorsal fin morphology and small cetacean behavior, the objective of this project was not met. However, lessons learned from these attempts to develop this new sampling tool have applicability for future researchers in the successful completion of a safe and effective methodology for remote attachment of satellite tags to small cetacean dorsal fins.

Exploring the Use of SEM-EDS Analysis to Measure the Distribution of Major, Minor, and Trace Elements in Bottlenose Dolphin (Tursiops truncatus) Teeth.

Dolphin teeth contain enamel, dentin, and cementum. In dentin, growth layer groups (GLGs), deposited at incremental rates (e.g., annually), are used for aging. Major, minor, and trace elements are incorporated within teeth; their distribution within teeth varies, reflecting tooth function and temporal changes in an individual's exposure. This study used a scanning electron microscope (SEM) equipped with energy dispersive X-ray spectroscopy (EDS) to determine the distribution of major (e.g., Ca, P), minor (e.g., Cl, Mg, Na), and trace elements (e.g., Cd, Hg, Pb, Zn) in teeth from 12 bottlenose dolphins (Tursiops truncatus). The objective was to compare elemental distributions between enamel and dentin and across GLGs. Across all dolphins and point analyses, the following elements were detected in descending weight percentage (wt %; mean ± SE): O (40.8 ± 0.236), Ca (24.3 ± 0.182), C (14.3 ± 0.409), P (14.0 ± 0.095), Al (4.28 ± 0.295), Mg (1.89 ± 0.047), Na (0.666 ± 0.008), Cl (0.083 ± 0.003). Chlorine and Mg differed between enamel and dentin; Mg increased from the enamel towards the dentin while Cl decreased. The wt % of elements did not vary significantly across the approximate location of the GLGs. Except for Al, which may be due to backscatter from the SEM stub, we did not detect trace elements. Other trace elements, if present, are below the detection limit. Technologies with lower detection limits (e.g., laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS)) would be required to confirm the presence and distribution of trace elements in bottlenose dolphin teeth.

Occurrence of Brucella ceti in stranded bottlenose dolphins Tursiops truncatus coincides with calving season.

Brucellosis is a disease caused by the Gram-negative facultative intracellular bacterium Brucella spp. In terrestrial species, this zoonotic bacterium is a global public health risk, but there is also concern over the zoonotic potential of marine forms, such as B. ceti, which affects cetaceans. Due to the detection of B. ceti in samples from bottlenose dolphins Tursiops truncatus during the 2010-2014 Gulf of Mexico Unusual Mortality Event, a long-term study of the prevalence of Brucella in stranded bottlenose dolphins from South Carolina, USA, was conducted. From 2012 through 2017, 282 stranded bottlenose dolphins were tested for B. ceti via real-time PCR. Nearly 32% of the dolphins tested positive in at least one sample (brain, lung, blowhole swab). Very little information exists in the literature on the occurrence of Brucella spp. in marine mammals, though in terrestrial species, such as cattle and elk, higher prevalence is often reported in spring. Similar results were found in this study with the peak occurrence being between March and June, a known period of calving in South Carolina. Results from this study provide important insights into the occurrence of the marine bacterium B. ceti.

Mercury concentrations in blubber and skin from stranded bottlenose dolphins (Tursiops truncatus) along the Florida and Louisiana coasts (Gulf of Mexico, USA) in relation to biological variables.

Due to their long life-span and top trophic position, odontocetes can accumulate high concentrations of mercury (Hg) in their tissues. This study measured the concentration of total Hg (THg) in the blubber and skin of bottlenose dolphins (Tursiops truncatus) that stranded along the Florida (FL) panhandle and Louisiana (LA) coasts and investigated the relationship between total Hg (THg) concentration and sex, body length, age, stranding location, diet/trophic position (δ13C and δ15N, respectively), and foraging habitat (δ34S). Additionally, we compared models using body length and age as explanatory variables to determine which was a better predictor of THg concentration. In both tissues, sex was not an influential predictor of THg concentration and there was a positive relationship between body length/age and THg concentration (p < 0.001). Florida dolphins had greater mean blubber and skin THg concentrations compared to LA dolphins (p < 0.001). There was a modest improvement in model fit when age was used in place of body length. δ13C, δ15N, and δ34S differed between stranding locations and together with age were significant predictors of THg concentrations (R2 = 0.52, P < 0.001). Florida dolphins were δ13C enriched compared to LA dolphins (p < 0.001) and THg concentrations were positively correlated with δ13C (R2 = 0.22, p < 0.001). Our results demonstrate spatial variability in THg concentrations from stranded bottlenose dolphins from the northern Gulf of Mexico; however, future research is required to understand how fine-scale population structuring of dolphins within FL and LA impacts THg concentrations, particularly among inshore (bay, sound, and estuary) stocks and between inshore and offshore stocks, as variations in biotic and abiotic conditions can influence both stable isotope ratios and THg concentrations.

Evidence for cortisol-cortisone metabolism by marine mammal blubber.

Blubber, a specialized hyperdermic adipose tissue found in marine mammals, has been identified as a useful tissue for the assessment of steroid hormone homeostasis in cetaceans. However, blubber cortisol measurements are not quantitatively predictive of circulating cortisol concentrations in bottlenose dolphins. In other mammals, adipose tissue metabolizes steroid hormones. Thus, it is proposed that the disagreement between blubber and blood cortisol in bottlenose dolphins could be due in part to metabolism of corticosteroids in blubber. The purpose of this study is to characterize the ability of blubber to interconvert cortisol and cortisone using an in vitro design. Results demonstrate that bottlenose dolphin blubber microsomes interconvert cortisol and cortisone, an effect that is abated by denaturing the microsomes, indicating this is an enzymatic process. These findings lead to the conclusion that blubber is likely a site of active steroid metabolism, which should be considered in future studies utilizing blubber as a matrix for endocrine assessment.

Selenium protein identification and profiling by mass spectrometry: A tool to assess progression of cardiomyopathy in a whale model.

Non-ischemic cardiomyopathy is a leading cause of congestive heart failure and sudden cardiac death in humans and in some cases the etiology of cardiomyopathy can include the downstream effects of an essential element deficiency. Of all mammal species, pygmy sperm whales (Kogia breviceps) present the greatest known prevalence of cardiomyopathy with more than half of examined individuals indicating the presence of cardiomyopathy from gross and histo-pathology. Several factors such as genetics, infectious agents, contaminants, biotoxins, and inappropriate dietary intake (vitamins, selenium, mercury, and pro-oxidants), may contribute to the development of idiopathic cardiomyopathy in K. breviceps. Due to the important role Se can play in antioxidant biochemistry and protein formation, Se protein presence and relative abundance were explored in cardiomyopathy related cases. Selenium proteins were separated and detected by multi-dimension liquid chromatography inductively coupled plasma mass spectrometry (LC-ICP-MS), Se protein identification was performed by liquid chromatography electrospray tandem mass spectrometry (LC-ESI-MS/MS), and Se protein profiles were examined in liver (n=30) and heart tissue (n=5) by SEC/UV/ICP-MS detection. Data collected on selenium proteins was evaluated in the context of individual animal trace element concentration, life history, and histological information. Selenium containing protein peak profiles varied in presence and intensity between animals with no pathological findings of cardiomyopathy and animals exhibiting evidence of cardiomyopathy. In particular, one class of proteins, metallothioneins, was found to be associated with Se and was in greater abundance in animals with cardiomyopathy than those with no pathological findings. Profiling Se species with SEC/ICP-MS proved to be a useful tool to identify Se protein pattern differences between heart disease stages in K. breviceps and an approach similar to this may be applied to other species to study Se protein associations with cardiomyopathy.

Rapid and reliable steroid hormone profiling in Tursiops truncatus blubber using liquid chromatography tandem mass spectrometry (LC-MS/MS).

Monitoring of marine mammal steroid hormone status using matrices alternative to blood is desirable due to the ability to remotely collect samples, which minimizes stress to the animal. However, measurement techniques in alternative matrices such as blubber described to date are limited in the number and types of hormones measured. Therefore, a new method using bead homogenization to QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) extraction, C18 post extraction cleanup and analysis by liquid chromatography tandem mass spectrometry (LC-MS/MS) was developed and applied to the measurement of hormone suites in bottlenose dolphin blubber. Validations were conducted in blubber from fresh dead stranded bottlenose dolphin. The final method consisting of two LC separations and garnet bead homogenization was tested for extraction efficiencies. Steroids were separated using a biphenyl column for reproductive hormones and C18 column for corticosteroids. Three hormones previously noted in blubber, testosterone, progesterone, and cortisol, were quantified in addition to previously unmeasured androstenedione, 17-hydroxyprogesterone, 11-deoxycortisol, 11-deoxycorticosterone, and cortisone in a single sample (0.4 g blubber). Extraction efficiencies of all hormones from blubber ranged from 84% to 112% and all RSDs were comparable to those reported using immunoassay methods (< 15%). The method was successfully applied to remote biopsied blubber samples to measure baseline hormone concentrations. Through this method, increased coverage of steroid hormone pathways from a single remotely collected sample potentially enhances the ability to interpret biological phenomena such as reproduction and stress in wild dolphin populations. Graphical abstract The steroid hormone profile is quantifiable from a single sample of bottlenose dolphin blubber using liquid chromatography tandem mass spectrometry. This profile can be applied to remotely collected dart biopsies and be used to determine reproductive or stress status of a wild-living dolphin.

Detection of Brucella spp. in bottlenose dolphins Tursiops truncatus by a real-time PCR using blowhole swabs.

Blowhole swabs are a simple and non-invasive method for collecting samples from cetaceans and can be used for screening large numbers of animals in the field. This study reports a real-time PCR assay for the detection of Brucella spp. using blowhole swab samples from bottlenose dolphins Tursiops truncatus stranded in the coastal region of Virginia, South Carolina and northern Florida, USA, between 2013 and 2015. We used real-time PCR results on lung samples from the same dolphins in order to estimate the relative sensitivity and specificity of real-time PCR of blowhole swabs. Brucella DNA was detected in lung tissue of 22% (18/81) and in blowhole swabs of 21% (17/81) of the sampled dolphins. The relative sensitivity and specificity of real-time PCR on blowhole swabs as compared to the real-time PCR on lung samples was 94% (17/18) and 100% (63/63), respectively. These results indicate that real-time PCR on blowhole swabs may be used as a non-invasive test for rapid detection of Brucella spp. in the respiratory tract of dolphins. To our knowledge, this is the first report on the use of blowhole swabs for detection of bacterial pathogens by real-time PCR in bottlenose dolphins.

A Bayesian mixture model for missing data in marine mammal growth analysis.

Much of what is known about bottle nose dolphin (Tursiops truncatus) anatomy and physiology is based on necropsies from stranding events. Measurements of total body length, total body mass, and age are used to estimate growth. It is more feasible to retrieve and transport smaller animals for total body mass measurement than larger animals, introducing a systematic bias in sampling. Adverse weather events, volunteer availability, and other unforeseen circumstances also contribute to incomplete measurement. We have developed a Bayesian mixture model to describe growth in detected stranded animals using data from both those that are fully measured and those not fully measured. Our approach uses a shared random effect to link the missingness mechanism (i.e. full/partial measurement) to distinct growth curves in the fully and partially measured populations, thereby enabling drawing of strength for estimation. We use simulation to compare our model to complete case analysis and two common multiple imputation methods according to model mean square error. Results indicate that our mixture model provides better fit both when the two populations are present and when they are not. The feasibility and utility of our new method is demonstrated by application to South Carolina strandings data.

A survey of trace element distribution in tissues of the dwarf sperm whale (Kogia sima) stranded along the South Carolina coast from 1990-2011.

Few studies report trace elements in dwarf sperm whale (Kogia sima). As high trophic level predators, marine mammals are exposed through diet to environmental contaminants including metals from anthropogenic sources. Inputs of Hg, Pb, and Cd are of particular concern due to toxicity and potential for atmospheric dispersion and subsequent biomagnification. Liver and kidney tissues of stranded K. sima from coastal South Carolina, USA, were analyzed for 22 trace elements. Age-related correlations with tissue concentrations were found for some metals. Mean molar ratio of Hg:Se varied with age with higher ratios found in adult males. Maximum concentrations of Cd and Hg in both tissues exceeded historical FDA levels of concern, but none exceeded the minimum 100µg/g Hg threshold for hepatic damage. Tissue concentrations of some metals associated with contamination were low, suggesting that anthropogenic input may not be a significant source of some metals for these pelagic marine mammals.

Real-time PCR assays for detection of Brucella spp. and the identification of genotype ST27 in bottlenose dolphins (Tursiops truncatus).

Rapid detection of Brucella spp. in marine mammals is challenging. Microbiologic culture is used for definitive diagnosis of brucellosis, but is time consuming, has low sensitivity and can be hazardous to laboratory personnel. Serological methods can aid in diagnosis, but may not differentiate prior exposure versus current active infection and may cross-react with unrelated Gram-negative bacteria. This study reports a real-time PCR assay for the detection of Brucella spp. and application to screen clinical samples from bottlenose dolphins stranded along the coast of South Carolina, USA. The assay was found to be 100% sensitive for the Brucella strains tested, and the limit of detection was 0.27fg of genomic DNA from Brucella ceti B1/94 per PCR volume. No amplification was detected for the non-Brucella pathogens tested. Brucella DNA was detected in 31% (55/178) of clinical samples tested. These studies indicate that the real-time PCR assay is highly sensitive and specific for the detection of Brucella spp. in bottlenose dolphins. We also developed a second real-time PCR assay for rapid identification of Brucella ST27, a genotype that is associated with human zoonotic infection. Positive results were obtained for Brucella strains which had been identified as ST27 by multilocus sequence typing. No amplification was found for other Brucella strains included in this study. ST27 was identified in 33% (18/54) of Brucella spp. DNA-positive clinical samples. To our knowledge, this is the first report on the use of a real-time PCR assay for identification of Brucella genotype ST27 in marine mammals.

Health of common bottlenose dolphins ( Tursiops truncatus ) in Barataria Bay, Louisiana, following the deepwater horizon oil spill.

The oil spill resulting from the explosion of the Deepwater Horizon drilling platform initiated immediate concern for marine wildlife, including common bottlenose dolphins in sensitive coastal habitats. To evaluate potential sublethal effects on dolphins, health assessments were conducted in Barataria Bay, Louisiana, an area that received heavy and prolonged oiling, and in a reference site, Sarasota Bay, Florida, where oil was not observed. Dolphins were temporarily captured, received a veterinary examination, and were then released. Dolphins sampled in Barataria Bay showed evidence of hypoadrenocorticism, consistent with adrenal toxicity as previously reported for laboratory mammals exposed to oil. Barataria Bay dolphins were 5 times more likely to have moderate-severe lung disease, generally characterized by significant alveolar interstitial syndrome, lung masses, and pulmonary consolidation. Of 29 dolphins evaluated from Barataria Bay, 48% were given a guarded or worse prognosis, and 17% were considered poor or grave, indicating that they were not expected to survive. Disease conditions in Barataria Bay dolphins were significantly greater in prevalence and severity than those in Sarasota Bay dolphins, as well as those previously reported in other wild dolphin populations. Many disease conditions observed in Barataria Bay dolphins are uncommon but consistent with petroleum hydrocarbon exposure and toxicity.

Ratiometric Measurements of Adiponectin by Mass Spectrometry in Bottlenose Dolphins (Tursiops truncatus) with Iron Overload Reveal an Association with Insulin Resistance and Glucagon.

High molecular weight (HMW) adiponectin levels are reduced in humans with type 2 diabetes and insulin resistance. Similar to humans with insulin resistance, managed bottlenose dolphins (Tursiops truncatus) diagnosed with hemochromatosis (iron overload) have higher levels of 2 h post-prandial plasma insulin than healthy controls. A parallel reaction monitoring assay for dolphin serum adiponectin was developed based on tryptic peptides identified by mass spectrometry. Using identified post-translational modifications, a differential measurement was constructed. Total and unmodified adiponectin levels were measured in sera from dolphins with (n = 4) and without (n = 5) iron overload. This measurement yielded total adiponectin levels as well as site specific percent unmodified adiponectin that may inversely correlate with HMW adiponectin. Differences in insulin levels between iron overload cases and controls were observed 2 h post-prandial, but not during the fasting state. Thus, post-prandial as well as fasting serum adiponectin levels were measured to determine whether adiponectin and insulin would follow similar patterns. There was no difference in total adiponectin or percent unmodified adiponectin from case or control fasting animals. There was no difference in post-prandial total adiponectin levels between case and control dolphins (mean ± SD) at 763 ± 298 and 727 ± 291 pmol/ml, respectively (p = 0.91); however, percent unmodified adiponectin was significantly higher in post-prandial cases compared to controls (30.0 ± 6.3 versus 17.0 ± 6.6%, respectively; p = 0.016). Interestingly, both total and percent unmodified adiponectin were correlated with glucagon levels in controls (r = 0.999, p < 0.001), but not in cases, which is possibly a reflection of insulin resistance. Although total adiponectin levels were not significantly different, the elevated percent unmodified adiponectin follows a trend similar to HMW adiponectin reported for humans with metabolic disorders.

Influence of mercury and selenium chemistries on the progression of cardiomyopathy in pygmy sperm whales, Kogia breviceps.

More than half of pygmy sperm whales (Kogia breviceps) that strand exhibit signs of cardiomyopathy (CMP). Many factors may contribute to the development of idiopathic CMP in K. breviceps, including genetics, infectious agents, contaminants, biotoxins, and dietary intake (e.g. selenium, mercury, and pro-oxidants). This study assessed trace elements in K. breviceps at various stages of CMP progression using fresh frozen liver and heart samples collected from individuals that stranded along US Atlantic and Gulf coasts between 1993 and 2007. Standard addition calibration and collision cell inductively coupled plasma mass spectrometry (ICP-MS) were employed for total Se analysis and pyrolysis atomic absorption (AA) was utilized for total Hg analysis to examine if the Se/Hg detoxification pathway inhibits the bioavailability of Se. Double spike speciated isotope dilution gas chromatography ICP-MS was utilized to measure methyl Hg and inorganic Hg. Immunoblot detection and colorimetric assays were used to assess protein oxidation status. Data collected on trace elements, selenoproteins, and oxidative status were evaluated in the context of animal life history and other complementary histological information to gain insight into the biochemical pathways contributing to the development of CMP in K. breviceps. Cardiomyopathy was only observed in adult pygmy sperm whales, predominantly in male animals. Both Hg:Se molar ratios and overall protein oxidation were greater in males than females and increased with progression of CMP.

Cryopreservation and in vitro culture of primary cell types from lung tissue of a stranded pygmy sperm whale (Kogia breviceps).

Current models for in vitro studies of tissue function and physiology, including responses to hypoxia or environmental toxins, are limited and rely heavily on standard 2-dimensional (2-D) cultures with immortalized murine or human cell lines. To develop a new more powerful model system, we have pursued methods to establish and expand cultures of primary lung cell types and reconstituted tissues from marine mammals. What little is known about the physiology of the deep-sea diving pygmy sperm whale (PSW), Kogia breviceps, comes primarily from stranding events that occur along the coast of the southeastern United States. Thus, development of a method for preserving live tissues and retrieving live cells from deceased stranded individuals was initiated. This report documents successful cryopreservation of PSW lung tissue. We established in vitro cultures of primary lung cell types from tissue fragments that had been cryopreserved several months earlier at the stranding event. Dissociation of cryopreserved lung tissues readily provides a variety of primary cell types that, to varying degrees, can be expanded and further studied/manipulated in cell culture. In addition, PSW-specific molecular markers have been developed that permitted the monitoring of fibroblast, alveolar type II, and vascular endothelial cell types. Reconstitution of 3-D cultures of lung tissues with these cell types is now underway. This novel system may facilitate the development of rare or disease-specific lung tissue models (e.g., to test causes of PSW stranding events and lead to improved treatments for pulmonary hypertension or reperfusion injury in humans). Also, the establishment of a "living" tissue bank biorepository for rare/endangered species could serve multiple purposes as surrogates for freshly isolated samples.

Anaemia, hypothyroidism and immune suppression associated with polychlorinated biphenyl exposure in bottlenose dolphins (Tursiops truncatus).

Polychlorinated biphenyls (PCBs), persistent chemicals widely used for industrial purposes, have been banned in most parts of the world for decades. Owing to their bioaccumulative nature, PCBs are still found in high concentrations in marine mammals, particularly those that occupy upper trophic positions. While PCB-related health effects have been well-documented in some mammals, studies among dolphins and whales are limited. We conducted health evaluations of bottlenose dolphins (Tursiops truncatus) near a site on the Georgia, United States coast heavily contaminated by Aroclor 1268, an uncommon PCB mixture primarily comprised of octa- through deca-chlorobiphenyl congeners. A high proportion (26%) of sampled dolphins suffered anaemia, a finding previously reported from primate laboratory studies using high doses of a more common PCB mixture, Aroclor 1254. In addition, the dolphins showed reduced thyroid hormone levels and total thyroxine, free thyroxine and triiodothyronine negatively correlated with PCB concentration measured in blubber (p = 0.039, < 0.001, 0.009, respectively). Similarly, T-lymphocyte proliferation and indices of innate immunity decreased with blubber PCB concentration, suggesting an increased susceptibility to infectious disease. Other persistent contaminants such as DDT which could potentially confound results were similar in the Georgia dolphins when compared with previously sampled reference sites, and therefore probably did not contribute to the observed correlations. Our results clearly demonstrate that dolphins are vulnerable to PCB-related toxic effects, at least partially mediated through the endocrine system. The severity of the effects suggests that the PCB mixture to which the Georgia dolphins were exposed has substantial toxic potential and further studies are warranted to elucidate mechanisms and potential impacts on other top-level predators, including humans, who regularly consume fish from the same marine waters.

Major pathologic findings and probable causes of mortality in bottlenose dolphins stranded in South Carolina from 1993 to 2006.

Although cause-of-death information on bottlenose dolphins (Tursiops truncatus) can be located in the literature, few citations include mortality data over a long period of time covering a broad geographic region. This study describes major pathologic findings and probable causes of death of bottlenose dolphins over a 14-yr period (1993-2006) for the coastal region of South Carolina. Probable causes of death for 97 cases were determined based on gross pathology and histopathology. In an additional 30 cases, probable cause of death was apparent from gross pathology alone, and carcass condition precluded histopathology. Of the 97 dolphins examined grossly and histologically, 30 (31%) likely died of infectious disease and 46 (47%) of noninfectious disease; the cause of death was unknown in 21 (22%). Bacterial infections accounted for the large majority of fatal infections and emaciation was the leading cause of noninfectious mortality. Twelve dolphins were killed by human interactions. Of the 30 dolphins diagnosed from gross examination alone, 23 likely died from human interaction and seven were killed by stingray-spine inflictions. Although the absence of consistent use of microbiology, biotoxin analysis and contaminant testing decreases the conclusiveness of the findings, this study has broad implications in establishing baseline data on causes of death of bottlenose dolphins for future studies and for the detection of emerging diseases.

Multiple congenital cardiac abnormalities in an Atlantic bottlenose dolphin (Tursiops truncatus).

Necropsy of an Atlantic bottlenose dolphin (Tursiops truncatus) neonate that stranded dead on Folly Beach, Charleston County, South Carolina, USA, on 17 November 2007, revealed multiple congenital heart malformations. Cardiac anomalies included a hypertrophic right ventricle, ventricular septal defect (VSD), aortic dilation, atrial septal defect (ASD) between a functionally common atrium and a left atrial remnant, subvalvular pulmonic stenosis, and a hypoplastic pulmonary artery and mitral valve. Few incidences of abnormal cardiac development in cetaceans have been published. The case study serves to document a novel congenital heart malformation not previously reported, to our knowledge, in free-ranging bottlenose dolphins.

Cytochrome P4501A1 expression, polychlorinated biphenyls and hydroxylated metabolites, and adipocyte size of bottlenose dolphins from the Southeast United States.

Persistent organic pollutants (POPs) bioaccumulate in blubber of marine mammals. Therefore, it is important to understand the structure and dynamics of blubber layers and how they affect the accumulation of POPs and subsequent biochemical responses. We used established histological and immunohistochemical methods to document the structure of bottlenose dolphin (Tursiops truncatus) blubber and to assess the expression of cytochrome P4501A1 (CYP1A1) in skin-blubber biopsies of dolphins sampled in the waters off Charleston, SC (CHS) (N=38), and Indian River Lagoon, FL (IRL) (N=36). CYP1A1 expression was strongest and most frequent in capillary endothelial cells and was stratified in blubber; the greatest CYP1A1 staining was in the deepest layer. CYP1A1 expression in deep blubber and 2,3,7,8-TCDD Toxic Equivalents measured in the entire blubber were significantly higher in dolphins from CHS as compared to those from IRL. Adipocyte size was associated with the extent of CYP1A1 expression. Male dolphins with smaller adipocytes from CHS and IRL had higher levels of CYP1A1 expression in deep blubber. In CHS females, CYP1A1 expression in vascular endothelial cells varied with reproductive status. CYP1A1 expression in the deep layer was highest in simultaneously pregnant-lactating dolphins, and these dolphins had the smallest adipocytes in deep blubber. In all dolphins, CYP1A1 expression in the deep blubber layer was positively related to concentrations of hydroxylated PCBs (OH-PCBs) in plasma. In summary, redistribution of AHR agonists from blubber into the circulatory system may enhance PCB metabolism and production of OH-PCBs by induction of CYP1A1 in hepatocytes and, possibly, by induction of CYP1A1 in endothelial cells of the deep blubber. The OH-PCBs thus formed have the potential to interfere with thyroid hormone homeostasis.