Lydia Staggs

A multi-year unusual mortality event (UME) involving primarily common bottlenose dolphins (Tursiops truncates) was declared in the northern Gulf of Mexico (GoM) with an initial start date of February 2010 and remains ongoing as of August 2014. To examine potential changing characteristics of the UME over time, we compared the number and demographics of dolphin strandings from January 2010 through June 2013 across the entire GoM as well as against baseline (1990-2009) GoM stranding patterns. Years 2010 and 2011 had the highest annual number of stranded dolphins since Louisiana's record began, and 2011 was one of the years with the highest strandings for both Mississippi and Alabama. Statewide, annual numbers of stranded dolphins were not elevated for GoM coasts of Florida or Texas during the UME period. Demographic, spatial, and temporal clusters identified within this UME included increased strandings in northern coastal Louisiana and Mississippi (March-May 2010); Barataria Bay, Louisiana (August 2010-December 2011); Mississippi and Alabama (2011, including a high prevalence and number of stranded perinates); and multiple GoM states during early 2013. While the causes of the GoM UME have not been determined, the location and magnitude of dolphin strandings during and the year following the 2010 Deepwater Horizon oil spill, including the Barataria Bay cluster from August 2010 to December 2011, overlap in time and space with locations that received heavy and prolonged oiling. There are, however, multiple known causes of previous GoM dolphin UMEs, including brevetoxicosis and dolphin morbillivirus. Additionally, increased dolphin strandings occurred in northern Louisiana and Mississippi before the Deepwater Horizon oil spill. Identification of spatial, temporal, and demographic clusters within the UME suggest that this mortality event may involve different contributing factors varying by location, time, and bottlenose dolphin populations that will be better discerned by incorporating diagnostic information, including histopathology.

Sixteen Chilean flamingos, Phoenicopterus chiles, and 10 red-tailed hawks, Buteo jamacensis, were vaccinated in the pectoral muscle with 0.2 ml of a commercially produced killed West Nile virus vaccine intended for use in horses. Half the birds of each species received a booster vaccination 3 weeks after the first injection. Three weeks after the booster vaccination, none of 13 birds surveyed had detectable antibody to West Nile virus.

Brucella ceti causes disease in Odontoceti. The absence of control serum collections and the diversity of cetaceans have hampered the standardization of serological tests for the diagnosis of cetacean brucellosis. Without a "gold" standard for sensitivity and specificity determination, an alternative approach was followed. We designed an indirect enzyme-linked immunosorbent assay (iELISA) that recognizes immunoglobulins G (IgGs) from 17 odontocete species as a single group. For the standardization, we used Brucella melitensis and Brucella abortus lipopolysaccharides, serum samples from seven resident odontocetes with no history of infectious disease displaying negative rose bengal test (RBT) reactions, and serum samples from seven dolphins infected with B. ceti. We compared the performance of the iELISA with those of the protein G ELISA (gELISA), the competitive ELISA (cELISA), and the immunofluorescence (IF) and dot blot (DB) tests, using 179 odontocete serum samples and RBT as the reference. The diagnostic potential based on sensitivity and specificity of the iELISA was superior to that of gELISA and cELISA. The correlation and agreement between the iELISA and the gELISA were relatively good (R(i/g)2 = 0.65 and kappa(i/g) = 0.66, respectively), while the correlation and agreement of these two ELISAs with cELISA were low (R(i/c)2 = 0.46, R(g/c)2 = 0.37 and kappa(i/c) = 0.62, kappa(g/c) = 0.42). In spite of using the same anti-odontocete IgG antibody, the iELISA was more specific than were the IF and DB tests. An association between high antibody titers and the presence of neurological symptoms in dolphins was observed. The prediction is that iELISA based on broadly cross-reacting anti-dolphin IgG antibody would be a reliable test for the diagnosis of brucellosis in odontocetes, including families not covered in this study.

The potential role of morbillivirus was evaluated in the deaths of >1100 bottlenose dolphins Tursiops truncatus and other small cetaceans that stranded from February 2010 through July 2014, during the northern Gulf of Mexico (GoM) unusual mortality event (UME). Morbillivirus analysis was carried out on 142 live or freshly dead cetaceans and results were combined with samples from 102 live, free-ranging bottlenose dolphins sampled during capture-release health assessments conducted from 2011 to 2014. Polymerase chain reaction (PCR) testing for morbillivirus showed that 9.9% (14/142) of the stranded cetaceans and 1% (1/83) of the free-ranging live dolphins were positive for dolphin morbilliviral (DMV) RNA. In contrast, previous DMV dolphin die-offs had DMV detectable by PCR in 61 to 97% of animals tested. Histologic findings consistent with morbillivirus infection, including lymphoid depletion, bronchointerstitial pneumonia, syncytial cell formation, or meningoencephalitis, were found in 6.6% (9/136) of the cetaceans that underwent histologic examinations. Serological analysis using a virus neutralization assay found that 29% (5/17) of live stranded and 23% (23/102) of live free-ranging bottlenose dolphins had titers of 64 or greater for cetacean morbillivirus, indicating prior but not necessarily recent exposure to morbillivirus. Current findings suggest that DMV infection, although present in the northern GoM, was sporadic and occurred at low levels and therefore was not the primary cause of the northern GoM UME. Confirmation of DMV infections and existing DMV titers demonstrate continued exposure to morbillivirus among northern GoM cetaceans since the first detection of this virus in the early 1990s.

Accurate identification of individual ages within wild bottlenose dolphins ( Tursiops truncatus ) is critical for determining population health and the development of population management strategies. As such, we analyzed DNA methylation (DNAm) patterns by applying a custom methylation array (HorvathMammalMethyl40) to both blood ( n = 140) and skin samples ( n = 87) from known age or approximate age (0–57 years) bottlenose dolphins. We present three bottlenose dolphin specific age estimation clocks using combined blood and skin [48 CpGs, R = 0.93, median absolute error (MAE) = 2.13 years], blood only (64 CpGs, R = 0.97, error = 1.46 years) and skin only (39 CpGs, R = 0.95, error = 2.53). We characterized individual cytosines that correlate with sex and age in dolphins and developed a sex estimator based on 71 CpGs that predicts the sex of any odontocete species with 99.5% accuracy. The presented epigenetic clocks are expected to be useful for conservation efforts and for determining if anthropogenic events affect aging rates in wild populations.