Patricia A. Fair

Occurrence of triclosan in plasma of wild Atlantic bottlenose dolphins (Tursiops truncatus) and in their environment.

The presence of triclosan, a widely-used antibacterial chemical, is currently unknown in higher trophic-level species such as marine mammals. Blood plasma collected from wild bottlenose dolphins (Tursiops truncatus) in Charleston, SC (CHS) (n = 13) and Indian River Lagoon, FL (IRL) (n = 13) in 2005 was analyzed for triclosan. Plasma concentrations in CHS dolphins ranged from 0.12 to 0.27 ng/g wet weight (mean 0.18 ng/g), with 31% of the sampled individuals having detectable triclosan. The mean IRL dolphin plasma concentrations were 0.072 ng/g wet weight (range 0.025-0.11 ng/g); 23% of the samples having detectable triclosan. In the CHS area, triclosan effluent values from two WWTP were both 190 ng/L and primary influents were 2800 ng/L and 3400 ng/L. Triclosan values in CHS estuarine surface water samples averaged 7.5 ng/L (n = 18) ranging from 4.9 to 14 ng/L. This is the first study to report bioaccumulation of anthropogenic triclosan in a marine mammal highlighting the need for further monitoring and assessment.

Geographic variation in polychorinated biphenyl and organochlorine pesticide concentrations in the blubber of bottlenose dolphins from the US Atlantic coast.

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 microg/g 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 1987/1988 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.

Contaminant blubber burdens in Atlantic bottlenose dolphins (Tursiops truncatus) from two southeastern US estuarine areas: Concentrations and patterns of PCBs, pesticides, PBDEs, PFCs, and PAHs

Polychlorinated biphenyls (PCBs), chlorinated pesticides (i.e., dichlorodiphenyltrichloroethane (DDT) and its metabolites, chlordanes (CHLs), dieldrin, hexachlorobenzene (HCB), and mirex), polybrominated diphenyl ethers (PBDEs), perfluorinated chemicals (PFCs), and polyaromatic hydrocarbons (PAHs) were measured in blubber biopsy samples collected from 139 wild bottlenose dolphins (Tursiops truncatus) during 2003-2005 in Charleston (CHS), SC and the Indian River Lagoon (IRL), FL. Dolphins accumulated a similar suite of contaminants with summation operatorPCB dominating (CHS 64%, IRL 72%), followed by summation operatorDDT (CHS 20%, IRL 17%), summation operatorCHLs (CHS 7%; IRL 7%), summation operatorPBDE (CHS 4%, IRL 2%), PAH at 2%, and dieldrin, PFCs and mirex each 1% or less. Together summation operatorPCB and summation operatorDDT concentrations contributed approximately 87% of the total POCs measured in blubber of adult males. summation operatorPCBs in adult male dolphins exceed the established PCB threshold of 17mg/kg by a 5-fold order of magnitude with a 15-fold increase for many animals; 88% of the dolphins exceed this threshold. For male dolphins, CHS (93,980ng/g lipid) had a higher summation operatorPCBs geomean compared to the IRL (79,752ng/g lipid) although not statistically different. In adult males, the PBDE geometric mean concentration was significantly higher in CHS (5920ng/g lipid) than the IRL (1487ng/g). Blubber summation operatorPFCs concentrations were significantly higher in CHS dolphins. In addition to differences in concentration of PCB congeners, summation operatorPBDE, TEQ, summation operatorCHLs, mirex, dieldrin, and the ratios summation operatorDDE/ summation operatorDDT and trans-nonachlor/cis-nonachlor were the most informative for discriminating contaminant loads in these two dolphin populations. Collectively, the current summation operatorPCB, summation operatorDDT, and summation operatorPBDEs blubber concentrations found in CHS dolphins are among the highest reported values in marine mammals. Both dolphin populations, particularly those in CHS, carry a suite of organic chemicals at or above the level where adverse effects have been reported in wildlife, humans, and laboratory animals warranting further examination of the potential adverse effects of these exposures.

Comparison of in vitro cytotoxicity, estrogenicity and anti‐estrogenicity of triclosan, perfluorooctane sulfonate and perfluorooctanoic acid

Concern with increasing levels of emerging contaminants exists on a global scale. Three commonly observed emerging environmental contaminants: triclosan (2,4,4‐trichloro‐2′‐hydroxydiphenyl ether), a synthetic, broad‐spectrum antibacterial agent, and perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA), used in stain‐ and water‐resistant treatments, have become distributed ubiquitously across ecosystems and have been detected in wildlife and humans. MCF‐7 BOS human breast cancer cells were used to investigate the potential for cytotoxicity, estrogenicity and anti‐estrogenicity of these three compounds at environmentally relevant concentrations using the 3‐(4,5‐dimethylthiazol‐2‐yl)‐5‐(3‐carboxymethoxyphenyl)‐2‐(4‐sulfophenyl)‐2H‐tetrazolium, inner salt assay (MTS) and the E‐SCREEN bioassay. The doses used were 0.002–200 µg ml−1 for triclosan and 0.03–30 µg ml−1 for PFOS and PFOA. Quantitative results from the MTS assay revealed no significant cytotoxicity at lower concentrations for any of the test compounds; however, both triclosan and PFOA were cytotoxic at the highest concentrations examined (100–200 and 30 µg ml−1, respectively), while PFOS showed no significant cytotoxicity at any of the concentrations tested. Positive estrogenic responses (P < 0.05) were elicited from the E‐SCREEN at all concentrations examined for triclosan and PFOA and at 30 µg ml−1 for PFOS. Further, significant anti‐estrogenic activity (P < 0.05) was detected for all compounds tested at all concentrations when cells were co‐exposed with 10−9 m 17‐β estradiol (E2). The overall results demonstrated that triclosan, PFOS and PFOA have estrogenic activities and that co‐exposure to contaminants and E2 produced anti‐estrogenic effects. Each of these compounds could provide a source of xenoestrogens to humans and wildlife in the environment. Published 2011. This article is a US Government work and is in the public domain in the USA.

Immune dysfunction in Atlantic bottlenose dolphins (Tursiops truncatus) with lobomycosis

Lobomycosis (Lacaziosis) occurs only in humans and dolphins under natural conditions. We evaluated the immune status of eight dolphins with lobomycosis and 40 healthy dolphins from the Indian River Lagoon (IRL), Florida. Lobomycosis cases had multiple abnormalities in their immunologic parameters when compared to healthy dolphins. The absolute number of circulating lymphocytes and serum albumin concentration were reduced (P<0.05) while the segmented neutrophils, alpha 1, total beta, total gamma and total globulins were increased (P<0.05). Although innate immunity was relatively intact and phagocytosis and natural killer cell activity were not affected, the plasma lysozyme concentrations were elevated in dolphins with lobomycosis (P<0.05). Adaptive immunity was depressed with statistically significant decreases found in the absolute numbers of CD4(+) helper T cells and CD19(+) and CD21(+) B cells. The ratios of CD2(+) T cells to CD4(+) cells and CD2(+) to CD21(+) cells were increased (P=0.05 and P<0.05, respectively) and the numbers of lymphocytes expressing MHC class II molecules was decreased in dolphins with lobomycosis (P<0.05). Lymphocyte proliferation was reduced in response to stimulation with lipopolysaccharide and concanavalin A (P<0.05). Antibody titers to Erysipelas rhusiopathiae, a common marine micro-organism, were decreased (P<0.05). In summary, dolphins with lobomycosis exhibit significant impairment in adaptive immunity.

Lobomycosis in Bottlenose Dolphins (Tursiops truncatus) from the Indian River Lagoon, Florida: Estimation of Prevalence, Temporal Trends, and Spatial Distribution

Lobomycosis (lacaziosis) is a chronic fungal disease of the skin that affects only dolphins and humans. Previous studies have shown a high prevalence of lobomycosis in bottlenose dolphins (Tursiops truncatus) from the Indian River Lagoon, Florida (IRL). We studied the occurrence and distribution of lobomycosis in the IRL using photo-identification survey data collected between 1996 and 2006. Our objectives were to (1) determine the sensitivity and specificity of photo-identification for diagnosis of lobomycosis in free-ranging dolphins; (2) determine the spatial distribution of lobomycosis in the IRL; and (3) assess temporal patterns of occurrence. Photographs from 704 distinctly marked dolphins were reviewed for skin lesions compatible with lobomycosis. The presumptive diagnosis was validated by comparing the results of photographic analysis with physical examination and histologic examination of lesion biopsies in 102 dolphins captured and released during a health assessment and 3 stranded dolphins. Twelve of 16 confirmed cases were identified previously by photography, a sensitivity of 75%. Among 89 dolphins without disease, all 89 were considered negative, a specificity of 100%. The prevalence of lobomycosis estimated from photographic data was 6.8% (48/704). Spatial distribution was determined by dividing the IRL into six segments based on hydrodynamics and geographic features. The prevalence ranged from <1% in the Mosquito Lagoon to 16.9% in the south Indian River. The incidence of the disease did not increase during the study period, indicating that the disease is endemic, rather than emerging. In summary, photo-identification is a useful tool to monitor the course of individual and population health for this enigmatic disease.

Correlation and toxicological inference of trace elements in tissues from stranded and free-ranging bottlenose dolphins (Tursiops truncatus).

The significance of metal concentrations in marine mammals is not well understood and relating concentrations between stranded and free-ranging populations has been difficult. In order to predict liver concentrations in free-ranging dolphins, we examined concentrations of trace elements (Al, As, Ba, Be, Cd, Co, Cu, Fe, Li, Mn, Ni, Pb, Sb, Se, Sn, total Hg (THg), V, Zn) in skin and liver of stranded bottlenose dolphins (Tursiops truncatus) from the South Carolina (SC) coast and the Indian River Lagoon, Florida (FL) during 2000-2008. Significantly higher concentrations of Zn, Fe, Se, Al, Cu and THg were found in skin while liver exhibited significantly higher Cu, Fe, Mn and THg concentrations for both study sites. Mean skin concentrations of Cu and Mn were significantly higher in SC dolphins while higher concentrations of THg and V were found in FL dolphins. In addition, liver tissues in SC dolphins exhibited significantly higher As concentrations while higher Fe, Pb, Se, THg, and V levels were found in FL dolphins. Two elements (Cu and THg) showed significant age-related correlations with skin concentration while five elements (Cu, Se, THg, Zn and V) showed age-related correlations with liver concentrations. Geographic location influenced age-related accumulation of several trace elements and age-related accumulation of THg in hepatic tissue was observed for both sites to have the highest correlations (r² = 0.90SC; r² = 0.69FL). Mean THg concentration in liver was about 10 times higher in FL dolphins (330 μg g⁻¹ dw) than those samples from SC dolphins (34.3 μg g⁻¹ dw). The mean molar ratio of Hg to Se was 0.93 ± 0.32 and 1.08 ± 0.38 for SC and FL dolphins, respectively. However, the Hg:Se ratio varied with age as much lower ratios (0.2-0.4) were found in younger animals. Of the 18 measured elements, only THg was significantly correlated in skin and liver of stranded dolphins and skin of free-ranging dolphins from both sites suggesting that skin may be useful in predicting Hg concentrations in liver tissue of free-ranging dolphins. Results indicate that 33% of the stranded and 15% of the free-ranging dolphins from FL exceed the minimum 100 μg g⁻¹ wet weight (ww) (~ 400 dw) Hg threshold for hepatic damage while none from SC reached this level. Hepatic concentrations of As in SC dolphins and V in FL dolphins were also highly correlated with skin concentrations which may have some regional specificity predictive value. The present study provides the first application of trace element concentrations derived from stranded bottlenose dolphins to predict liver concentrations in free-ranging populations.

Hematologic, biochemical, and cytologic findings from apparently healthy atlantic bottlenose dolphins (Tursiops truncatus) inhabiting the Indian River Lagoon, Florida, USA.

The objective of this study was to establish reference baseline data for hematologic, biochemical, and cytologic findings in apparently healthy Atlantic bottlenose dolphins (Tursiops truncatus) inhabiting the Indian River Lagoon, Florida, USA. Sixty-two dolphins were captured, examined, and released during June 2003 and June 2004. Mean, standard deviation, and range were calculated for each parameter, and values for which published data were available, were close to or within the ranges previously reported for free-ranging bottlenose dolphins. No pathologic abnormalities were found in fecal and blowhole cytologic specimens. However, 24% (7/29) of the dolphins examined in 2003 had evidence of gastritis, which was graded as severe in 14% (4/29) of the cases. In 2004, only 4% (1/24) of dolphins sampled had evidence of mild or moderate gastritis; no severe inflammation was present. Dolphins with evidence of gastritis were 8 yr of age or older and predominantly male. Several statistically significant differences were found between males and females, between pregnant and nonpregnant animals, and between juveniles (<6 yr) and adults (≥6 yr). However, the values remained within the established ranges for this species, and the differences were not likely to be of clinical significance.

Evaluation and comparison of the health status of Atlantic bottlenose dolphins from the Indian River Lagoon, Florida, and Charleston, South Carolina

OBJECTIVE To conduct health assessments and compare outcomes in 2 populations of Atlantic bottlenose dolphins. Design-Repeated cross-sectional study. ANIMALS 171 Atlantic bottlenose dolphins. PROCEDURES During June and August of 2003 through 2005, 89 dolphins from the Indian River Lagoon (IRL), Florida, and 82 dolphins from estuarine waters near Charleston, SC, were evaluated. A panel of 5 marine mammal veterinarians classified dolphins as clinically normal, possibly diseased, or definitely diseased on the basis of results of physical and ultrasonographic examinations, hematologic and serum biochemical analyses, and cytologic and microbiologic evaluations of gastric contents and swab specimens. RESULTS Prevalence of dolphins classified as definitely diseased did not differ significantly between the IRL (32%) and Charleston (20%) sites. Proportions of dolphins classified as possibly diseased also did not differ. Lobomycosis was diagnosed in 9 dolphins from the IRL but in none of the dolphins from Charleston. Proportions of dolphins with orogenital papillomas did not differ significantly between the IRL (12%) and Charleston (7%) sites. From 2003 through 2005, the proportion classified as definitely diseased tripled among dolphins from the Charleston site but did not increase significantly among dolphins from the IRL. Dolphins from the Charleston site were more likely to have leukocytosis, lymphocytosis, and low serum concentrations of total protein and total J-globulins than were dolphins from the IRL. CONCLUSIONS AND CLINICAL RELEVANCE High prevalences of diseased dolphins were identified at both sites; however, the host or environmental factors that contributed to the various abnormalities detected are unknown.

Orogenital Neoplasia in Atlantic Bottlenose Dolphins (Tursiops truncatus)

This study describes lingual papillomas and squa- mous cell carcinomas (n = 11) and genital pap- illomas (n = 4) in Atlantic bottlenose dolphins (Tursiops truncatus) evaluated from January 2000 to January 2005. Tumors were found primarily in adult dolphins of both sexes living in free-ranging and captive conditions. Three dolphins had mul- tiple lingual tumors of mixed histological type, consisting of papillomas and squamous cell car- cinomas, suggesting malignant transformation of the benign papillomatous lesions. To our knowl- edge, this is the first report of oral papillomas in bottlenose dolphins and concurrent oral neoplasia that included both sessile papilloma and squamous cell carcinoma in the same dolphin. Additionally, it is the first known report of genital papillomas in free-ranging bottlenose dolphins from Atlantic coastal waters. The unusually high occurrence of related benign and malignant orogenital epithelial neoplastic lesions in a short period suggests that the lesions may represent one or more emerging diseases. Preliminary evidence suggests that these tumors may be of infectious etiology, possibly having an orogenital route of transmission.