Wayne E. McFee

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.

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.

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.

TWO NOVEL ALPHAHERPESVIRUSES ASSOCIATED WITH FATAL DISSEMINATED INFECTIONS IN ATLANTIC BOTTLENOSE DOLPHINS

Two immature female bottlenose dolphins (Tursiops truncatus) were found stranded on the Atlantic coast of the USA. Necropsy and histopathologic examination of both dolphins demonstrated acute necrotizing lesions in multiple organ systems. Commonly seen in these lesions were cells with enlarged nuclei that contained single 4 to 6 μm diameter homogeneous eosinophilic inclusion bodies that were often surrounded by a clear halo. Ultrastructural examination revealed that intranuclear inclusions contained 90 to 110 nm diameter viral particles with electron-dense cores and hexagonal profiles. Viral particles were also present in the cytoplasm, and these were surrounded by variably electron-dense envelopes. Enveloped virions were 140 nm in diameter. Polymerase chain reactions targeting the DNA polymerase and terminase genes of herpesviruses were carried out on unfixed tissues of both animals, and analysis of the DNA products indicated the presence of two novel alphaherpesviruses. The gross, histologic, ultrastructural, and molecular genetic findings indicate disseminated herpesviral infections, and support the conclusion that the alphaherpesviruses caused the deaths of the two dolphins. This is the first report of disseminated herpesviral infection in cetaceans.

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.

Factors influencing persistent organic pollutant concentrations in the Atlantic white-sided dolphin (Lagenorhynchus acutus).

Assessing the trends of persistent organic pollutants (POPs) in cetaceans is difficult because of age and gender influences on accumulation. Persistent organic pollutants bioaccumulate and are poorly metabolized; hence, concentrations may increase with age in males while females reduce their POP burden through parturition and lactation. Age and gender effects on contaminant concentrations are species specific because of life history and reproductive strategies. These influences must be understood in order to elucidate and assess lifetime POP exposure. The objectives of this study were to determine baseline POP concentrations in blubber samples from the Atlantic white-sided dolphin (Lagenorhynchus acutus) and to investigate life history and other influences, such as metabolism, on these concentrations. Forty-seven L. acutus blubber samples collected from mass stranding events in Massachusetts, USA (1993-2000), and archived in the National Marine Mammal Tissue Bank at the National Institute of Standards and Technology (Gaithersburg, MD, USA) were analyzed for 55 polychlorinated biphenyl congeners (PCB; 55 congeners), five polybrominated diphenyl ether (PBDE) congeners, and organochlorine pesticides (toxaphene, DDT and metabolites, mirex, dieldrin, chlordanes, hexachlorocyclohexanes, hexachlorobenzene, and endosulfans) by gas chromatography/mass spectrometry (GC-MS). Ages for 19 animals were determined from growth layer groups on decalcified, stained thin tooth sections. Total PCBs (sigmaPCB; sum of 55 congeners) were the contaminants present in the highest concentrations in all age classes (0.5-63 microg/g wet mass) followed by sum of DDTs (0.50-43 microg/g wet mass), toxaphene (0.055-31 microg/g wet mass), chlordanes (0.30-24 microg/g wet mass), and PBDEs (0.12-4.0 microg/g wet mass). Body length had a greater statistical influence than age on contaminant burdens in L. acutus. Contaminant burdens decreased with length in both male and female L. acutus, suggesting metabolic elimination and/or growth dilution in males and off-loading via lactation in females.

Skin Lesions on Common Bottlenose Dolphins (Tursiops truncatus) from Three Sites in the Northwest Atlantic, USA

Skin disease occurs frequently in many cetacean species across the globe; methods to categorize lesions have relied on photo-identification (photo-id), stranding, and by-catch data. The current study used photo-id data from four sampling months during 2009 to estimate skin lesion prevalence and type occurring on bottlenose dolphins (Tursiops truncatus) from three sites along the southeast United States coast [Sarasota Bay, FL (SSB); near Brunswick and Sapelo Island, GA (BSG); and near Charleston, SC (CHS)]. The prevalence of lesions was highest among BSG dolphins (P = 0.587) and lowest in SSB (P = 0.380), and the overall prevalence was significantly different among all sites (p<0.0167). Logistic regression modeling revealed a significant reduction in the odds of lesion occurrence for increasing water temperatures (OR = 0.92; 95%CI:0.906–0.938) and a significantly increased odds of lesion occurrence for BSG dolphins (OR = 1.39; 95%CI:1.203–1.614). Approximately one-third of the lesioned dolphins from each site presented with multiple types, and population differences in lesion type occurrence were observed (p<0.05). Lesions on stranded dolphins were sampled to determine the etiology of different lesion types, which included three visually distinct samples positive for herpesvirus. Although generally considered non-fatal, skin disease may be indicative of animal health or exposure to anthropogenic or environmental threats, and photo-id data provide an efficient and cost-effective approach to document the occurrence of skin lesions in free-ranging populations.

Fetal distress and in utero pneumonia in perinatal dolphins during the Northern Gulf of Mexico unusual mortality event.

An unusual mortality event (UME) involving primarily common bottlenose dolphins Tursiops truncatus of all size classes stranding along coastal Louisiana, Mississippi, and Alabama, USA, started in early 2010 and continued into 2014. During this northern Gulf of Mexico UME, a distinct cluster of perinatal dolphins (total body length <115 cm) stranded in Mississippi and Alabama during 2011. The proportion of annual dolphin strandings that were perinates between 2009 and 2013 were compared to baseline strandings (2000-2005). A case-reference study was conducted to compare demographics, histologic lesions, and Brucella sp. infection prevalence in 69 UME perinatal dolphins to findings from 26 reference perinates stranded in South Carolina and Florida outside of the UME area. Compared to reference perinates, UME perinates were more likely to have died in utero or very soon after birth (presence of atelectasis in 88 vs. 15%, p < 0.0001), have fetal distress (87 vs. 27%, p < 0.0001), and have pneumonia not associated with lungworm infection (65 vs. 19%, p = 0.0001). The percentage of perinates with Brucella sp. infections identified via lung PCR was higher among UME perinates stranding in Mississippi and Alabama compared to reference perinates (61 vs. 24%, p = 0.01), and multiple different Brucella omp genetic sequences were identified in UME perinates. These results support that from 2011 to 2013, during the northern Gulf of Mexico UME, bottlenose dolphins were particularly susceptible to late-term pregnancy failures and development of in utero infections including brucellosis.

Papillomavirus antibody prevalence in free-ranging and captive bottlenose dolphins (Tursiops truncatus).

Genital epithelial tumors of Atlantic bottlenose dolphins (Tursiops truncatus [Tt]) and Burmeisters porpoises (Phocoena spinipinnis) were formerly shown to be associated with papillomavirus (PV) infection. Papillomaviruses are highly prevalent viruses involved in the development of various tumor types in a wide range of animals, and so-called high-risk PVs contribute to malignant progression. In marine mammals, the incidence and prevalence of PV infection, transmission pathways, and persistence of infection are largely unknown. Using virus-like particles of bottlenose dolphin PV type 1 (TtPV1) as the antigen, enzyme-linked immunosorbent assay (ELISA) studies were conducted to evaluate PV antibody prevalence in bottlenose dolphins. In total, sera obtained from 115 dolphins were examined. Fifty-one percent of captive dolphins (n=18 of 35) and 90% of free-ranging dolphins (n=72 of 80) were antibody positive. Higher ELISA reactivity was observed among males compared with females. Sexually immature dolphins appeared more likely to seroconvert with age. Besides determining their PV antibody prevalence, each animal was also assessed for the presence of orogenital tumors. Interestingly, the mean age of free-ranging dolphins with tumors (n=21) was 11.2 yr compared with 29.9 yr in captive dolphins with tumors (n=9). Results from the current study suggest PV infection in bottlenose dolphins is common, that the main route of PV transmission among them may be horizontal, and that orogenital neoplasia may develop in early life stages of certain free-ranging bottlenose dolphins.

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.