Clonally related methicillin-resistant Staphylococcus aureus isolated from short-finned pilot whales (Globicephala macrorhynchus), human volunteers, and a bayfront cetacean rehabilitation facility.

In May of 2011, a live mass stranding of 26 short-finned pilot whales (Globicephala macrorhynchus) occurred in the lower Florida Keys. Five surviving whales were transferred from the original stranding site to a nearby marine mammal rehabilitation facility where they were constantly attended to by a team of volunteers. Bacteria cultured during the routine clinical care of the whales and necropsy of a deceased whale included methicillin-sensitive and methicillin-resistant Staphylococcus aureus (MSSA and MRSA). In order to investigate potential sources or reservoirs of MSSA and MRSA, samples were obtained from human volunteers, whales, seawater, and sand from multiple sites at the facility, nearby recreational beaches, and a canal. Samples were collected on 3 days. The second collection day was 2 weeks after the first, and the third collection day was 2 months after the last animal was removed from the facility. MRSA and MSSA were isolated on each day from the facility when animals and volunteers were present. MSSA was found at an adjacent beach on all three collection days. Isolates were characterized by utilizing a combination of quantitative real-time PCR to determine the presence of mecA and genes associated with virulence, staphylococcal protein A typing, staphylococcal cassette chromosome mec typing, multilocus sequence typing, and pulsed field gel electrophoresis (PFGE). Using these methods, clonally related MRSA were isolated from multiple environmental locations as well as from humans and animals. Non-identical but genetically similar MSSA and MRSA were also identified from distinct sources within this sample pool. PFGE indicated that the majority of MRSA isolates were clonally related to the prototype human strain USA300. These studies support the notion that S. aureus may be shed into an environment by humans or pilot whales and subsequently colonize or infect exposed new hosts.

Vulnerability to climate change of United States marine mammal stocks in the western North Atlantic, Gulf of Mexico, and Caribbean.

Climate change and climate variability are affecting marine mammal species and these impacts are projected to continue in the coming decades. Vulnerability assessments provide a framework for evaluating climate impacts over a broad range of species using currently available information. We conducted a trait-based climate vulnerability assessment using expert elicitation for 108 marine mammal stocks and stock groups in the western North Atlantic, Gulf of Mexico, and Caribbean Sea. Our approach combined the exposure (projected change in environmental conditions) and sensitivity (ability to tolerate and adapt to changing conditions) of marine mammal stocks to estimate vulnerability to climate change, and categorize stocks with a vulnerability index. The climate vulnerability score was very high for 44% (n = 47) of these stocks, high for 29% (n = 31), moderate for 20% (n = 22), and low for 7% (n = 8). The majority of stocks (n = 78; 72%) scored very high exposure, whereas 24% (n = 26) scored high, and 4% (n = 4) scored moderate. The sensitivity score was very high for 33% (n = 36) of these stocks, high for 18% (n = 19), moderate for 34% (n = 37), and low for 15% (n = 16). Vulnerability results were summarized for stocks in five taxonomic groups: pinnipeds (n = 4; 25% high, 75% moderate), mysticetes (n = 7; 29% very high, 57% high, 14% moderate), ziphiids (n = 8; 13% very high, 50% high, 38% moderate), delphinids (n = 84; 52% very high, 23% high, 15% moderate, 10% low), and other odontocetes (n = 5; 60% high, 40% moderate). Factors including temperature, ocean pH, and dissolved oxygen were the primary drivers of high climate exposure, with effects mediated through prey and habitat parameters. We quantified sources of uncertainty by bootstrapping vulnerability scores, conducting leave-one-out analyses of individual attributes and individual scorers, and through scoring data quality for each attribute. These results provide information for researchers, managers, and the public on marine mammal responses to climate change to enhance the development of more effective marine mammal management, restoration, and conservation activities that address current and future environmental variation and biological responses due to climate change.

Macroscopic and Histopathologic Findings From a Mass Stranding of Rough-Toothed Dolphins (Steno bredanensis) in 2005 on Marathon Key, Florida, USA.

On March 2, 2005 ~70 rough-toothed dolphins (Steno bredanensis) mass stranded along mud flats and associated canals on the Atlantic Ocean side of Marathon Key, Florida. Forty-six were necropsied and placed into two groups for analysis: Group-1 animals (N = 34; 65%) that died prior to medical intervention and rehabilitative efforts and Group-2 animals (N = 12; 35%) that died in rehabilitation. Thirty-four animals were females (18 adults, 5 juvenile/subadult, 7 calves, and 4 of undetermined age) and 12 were males (6 adults, 4 juvenile/subadults, 1 calf, and 1 of undetermined age). Body condition overall was fair to good in Group-1 and fair to poor in Group-2. Lesions were observed in multiple body systems. Greater than 90% of animals in both groups had respiratory lesions. Verminous sinusitis and bronchopneumonia were 2-3 times more prevalent in Group-2. Capture/exertional rhabdomyolysis was observed in Group-2 (42%). Vacuolar hepatopathies were observed in both groups including hepatic lipidosis (Group-1) and mixed etiologies (Group-2). Pancreatic and gastrointestinal tract pathologies were prevalent in Group-2 animals 56 and 75%, respectively, and included gastritis, gastric ulceration, enterocolitis, pancreatic atrophy, and pancreatitis related to physiologic stress. Group-2 more frequently had evidence of hemorrhagic diathesis present which included increased extramedullary hematopoiesis in various organs, increased hemosiderosis, and hemorrhage and hemorrhagic drainage in various organs. Central nervous system disease, primarily edema, and mild inflammation were equally prevalent. Renal proteinuria, tubular necrosis, and pigmentary deposition were observed in Group-2. Dental attrition was observed in ~40% of the groups. Gammaherpesviral-associated pharyngeal plaques were observed in 46 and 54% of Group-1 and 2 animals, respectively. Other lesions observed were mild and incidental with a frequency rate <20%. The findings from this Steno stranding provide a unique window into baseline individual and population clinical conditions and additional perspective into potential clinical sequelae of rehabilitation efforts.

Auditory evoked potentials in a stranded Gervais' beaked whale (Mesoplodon europaeus).

Efforts to identify the specific causal mechanisms responsible for beaked whale strandings coincident with naval exercises have been hampered by lack of data concerning the hearing abilities of beaked whales and their physiological and behavioral responses to sound. In this study, auditory capabilities of a stranded Gervais' beaked whale (Mesoplodon europaeus) were investigated by measuring auditory evoked potentials. Click-evoked potentials, auditory thresholds as a function of frequency, and the modulation rate transfer function were determined. The evoked potentials and modulation rate transfer function were similar to those measured in other echolocating odontocetes; the upper limit of functional hearing was 80-90 kHz.

Evaluation of body mass index as a prognostic indicator from two rough-toothed dolphin (Steno bredanensis) mass strandings in Florida.

Rough-toothed dolphins (Steno bredanensis) are a common mass stranding species in Florida. These large stranding events typically include a small number of sick or injured individuals and a much larger number of healthy individuals, making rapid triage essential. Little data exist on rehabilitation outcomes, and historically, successful outcomes are limited. Furthermore, very little data exist on the feeding habits and dietary needs of this species. This study compared morphology and body mass index (BMI) in two rough-toothed dolphin mass stranding events in Florida: August 2004 (n = 36) and March 2005 (n = 32). The two groups were significantly different in morphologic measurements, with age and gender-adjusted intake BMI significantly (p < .01) different (2004 = 0.34 ± 0.02; 2005 = 0.41 ± 0.02) between groups. Ten animals from 2005 had weights tracked throughout the rehabilitation process and demonstrated an initial drop in BMI followed by an increase and a plateau prior to release. When comparing initial BMI by stranding outcome, individuals that were rehabilitated and released had a significantly (p = .03) higher BMI than individuals who were euthanized. However, there was no difference between dolphins that died of natural causes (p = .56) and animals successfully rehabilitated. Analysis of BMI can be a useful marker in triage during a stranding, when resources are limited to identify individuals most likely to survive, as well as in determining the appropriate body condition for release. The data reported here can provide guidance on evaluating the nutritive status on this uncommon species that would otherwise be difficult to obtain among wild populations.

Evaluation of Serum for Pathophysiological Effects of Prolonged Low Salinity Water Exposure in Displaced Bottlenose Dolphins (Tursiops truncatus).

We conducted a retrospective study of serum biochemistry and hematologic findings from displaced, out-of-habitat bottlenose dolphins (Tursiops truncatus) exposed to various low salinity environments in waters along the southern United States including southeastern Atlantic and northern Gulf of Mexico. Serum sodium, chloride, and calculated osmolality were significantly lower and below reference ranges in displaced animals compared to free-ranging case control animals. This suggests clinical hyponatremia, hypochloremia, and hypo-osmolality due to an uptake of low saline water from the environment. In addition, significant differences were found in other serum chemistry variables, although none were outside of normal reference ranges for non-controlled free-ranging animals. Multiple linear regressions demonstrated the degree of salinity had a greater pathophysiologic response than the duration of fresh water exposure. The Na/Cl ratio and bicarbonate were the only variables that were significantly modulated by exposure duration. These findings suggest that the degree of salinity is a critical factor when assessing and managing care for dolphins chronically exposed to low salinity water. Results from this study indicate that changes in various biochemical parameters can be used to determine fresh water exposure and aid in determining the treatment for animals recovered from low salinity waters.

Identification of novel alpha- and gammaherpesviruses from cutaneous and mucosal lesions of dolphins and whales.

Total DNA extracted from mucosal and skin lesions of captive and stranded cetaceans was analyzed for herpesvirus DNA by nested and direct polymerase chain reactions (PCR). The targeted sequences corresponded to a region of the DNA polymerase gene containing multiple conserved amino acid motifs. Herpesvirus genomic DNA fragments (222-244 bp) were amplified from 11 lesions by nested PCR and from eight lesions ( approximately 730 bp) using direct PCR from US cetaceans. Fragments of various sizes were also amplified from skin, spleen and blood of a German dolphin. Sequencing and BLAST analysis of these DNA fragments indicated that alpha- or gammaherpesviruses were present in the cetacean lesions. Alphaherpesviruses were associated with skin lesions of three Atlantic bottlenose dolphins (Tursiops truncatus), while gammaherpesviruses were present in genital lesions of five Atlantic bottlenose dolphins, one Risso's dolphin (Grampus griseus), one dwarf sperm whale (Kogia sima) and one Blainville's beaked whale (Mesoplodon densirostris), as well as in one oral lesion from an Atlantic bottlenose dolphin. Phylogenetic analysis of deduced amino acid sequences showed that the cetacean alphaherpesviruses were most closely related to human alphaherpesviruses, namely, herpes simplex-1 and -2. On the other hand, cetacean gammaherpesviruses were most closely related to Rhadinoviruses. These novel cetacean herpesviruses appeared to be distinct from known herpesviruses of marine and terrestrial vertebrates. The sequencing data strongly suggest that these viruses are most likely cetacean specific and possibly have coevolved with their cetacean hosts.

Neurological disease in wild loggerhead sea turtles Caretta caretta.

Beginning in October 2000, subadult loggerhead sea turtles Caretta caretta showing clinical signs of a neurological disorder were found in waters off south Florida, USA. Histopathology indicated generalized and neurologic spirorchiidiasis. In loggerhead sea turtles (LST) with neurospirorchiidiasis, adult trematodes were found in the meninges of the brain and spinal cord of 7 and 3 affected turtles respectively, and multiple encephalic intravascular or perivascular eggs were associated with granulomatous or mixed leukocytic inflammation, vasculitis, edema, axonal degeneration and occasional necrosis. Adult spirorchiids were dissected from meningeal vessels of 2 of 11 LST brains and 1 of 10 spinal cords and were identified as Neospirorchis sp. Affected LST were evaluated for brevetoxins, ciguatoxins, saxitoxins, domoic acid and palytoxin. While tissues from 7 of 20 LST tested positive for brevetoxins, the levels were not considered to be in a range causing acute toxicosis. No known natural (algal blooms) or anthropogenic (pollutant spills) stressors co-occurred with the turtle mortality. While heavy metal toxicosis and organophosphate toxicosis were also investigated as possible causes, there was no evidence for their involvement. We speculate that the clinical signs and pathologic changes seen in the affected LST resulted from combined heavy spirorchiid parasitism and possible chronic exposure to a novel toxin present in the diet of LST.

A poorly differentiated pulmonary squamous cell carcinoma in a free-ranging Atlantic bottlenose dolphin (Tursiops truncatus).

A free-ranging, adult, female offshore bottlenose dolphin (Tursiops truncatus) was found freshly dead in 1999 on Ocean Park Beach in San Juan, Puerto Rico. The left-lung and right-lung pleura had multiple white, firm-to-hard nodules with coagulative necrosis. Histologically, the neoplasms were characterized by multiple well-circumscribed, nonencapsulated expansile masses consisting mostly of polygonal cells with fewer circumferential flattened basaloid cells that compressed alveoli, bronchioles, and bronchi. Neoplastic cells stained positive for cytokeratin, with sporadic vimentin staining, and were negative for epithelial membrane antigen, thyroid transcription factor-1, calretinin, and human mesothelial cell antigen. A diagnosis of poorly differentiated pulmonary squamous cell carcinoma with lymph node and renal metastases was made on the basis of histomorphology and immunohistochemical staining. This is the first documentation of pulmonary squamous cell carcinoma in a dolphin.

Viral papillomatosis in Florida manatees (Trichechus manatus latirostris).

The Florida manatee (Trichechus manatus latirostris) is one of the most endangered marine mammals in American coastal waters. Naturally resistant to infectious disease, the manatee immune system appears highly developed to protect it against the harsh marine environment and the effects of human-related injury. In 1997, seven captive Florida manatees developed multiple, cutaneous, pedunculated papillomas over a period of 6 months. Approximately 3 years later, four of the seven manatees developed multiple, cutaneous, sessile papillomas topically and clinically distinct from the initial lesions, some of which are still present. Histologic, ultrastructural, and immunohistochemical features indicated that the two distinct phenotypic lesions were caused by papillomaviruses (PVs). Preliminary immunologic data correlated with daily clinical observations suggested that the manatees were immunologically suppressed and that the papillomas were caused by activation of latent PV infections and reinoculation from active infections. The emergence of PV-induced papillomas in captive manatees, the possibility of activation of latent infection or transmission of active infection to free-ranging manatees, and the underlying cause of immune suppression predisposing manatees to develop viral papillomatosis are serious concerns for the future management of this highly endangered species.