Dean A. Bagley

Internesting and Postnesting Movements and Foraging Habitats of Leatherback Sea Turtles (Dermochelys coriacea) Nesting in Florida

ABSTRACT We tracked 10 leatherback turtles by satellite from 2 Florida Atlantic Coast nesting beaches for a period ranging from 38 days to more than 454 days. Movement and foraging areas were often coastal, which contrasts with other satellite telemetry studies where leatherbacks are more pelagic. Using kernel home-range estimation we identified the primary internesting residence areas as well as Atlantic foraging areas. The primary internesting habitat was centered east–southeast of Cape Canaveral, Florida, from 2 to 60 km offshore and extending 215 km along the coast. Atlantic foraging areas were located primarily on the continental shelf from 30° to 50°N, and in an offshore area centered at 42°N, 65°W, as well as off Africa in the Mauritania upwelling. Seasonally, the location of these foraging areas changed, occurring on the North American continental shelf from March through November and off the shelf from December through February. One of the tracked turtles may have been killed with 17 other leatherbacks by coastal shrimp fishing located near the Georgia–Florida border. We illustrate how using remotely sensed data could be used to prevent such mortalities.

Spatiotemporal patterns of annual sea turtle nesting behaviors along an East Central Florida beach

Abstract The Florida coastline from Melbourne Beach to Wabasso Beach is one the most important nesting areas for loggerhead turtles ( Caretta caretta ) in the Western Hemisphere and for green turtles ( Chelonia mydas ) in the United States. In this study, we quantified the spatial patterns of numerous loggerhead ( N ≈400,000) and green turtle ( N ≈14,000) and less numerous ( N ≈100) leatherback ( Dermochelys coriacea ) beach ascents from 1989–1999 in terms of their autocorrelative properties along 40.5 km within this critical reproductive zone. Nesting and non-nesting emergence patterns of loggerhead and green turtles were non-random, favoring the southern half of the study area. Perhaps due to low numbers or differences in nesting behavior, leatherback nest distributions were not significantly different from random. Loggerhead and green turtle nest locations exhibited similar clinal patterns. They were positively autocorrelated at distances less than 10 km and negatively at distances greater than 30 km. These patterns were significantly correlated interannually.

Serological association between spirorchidiasis, herpesvirus infection, and fibropapillomatosis in green turtles from Florida.

Serodiagnostic tests for detecting green turtle (Chelonia mydas) antibody responses were developed to test the strength of association between exposure to spirorchid trematode antigens or herpesvirus antigens and having green turtle fibropapillomatosis (GTFP). Plasma samples from 46 captive-reared green turtles, including paired pre- and 1-yr post-inoculation samples from 12 turtles with experimentally induced GTFP, were found by enzyme-linked immunosorbent assay (ELISA) to be negative for antibodies to adult spirorchid (Learedius learedi) antigens. In contrast, all 12 turtles that developed experimentally induced GTFP converted within 1 yr from having negative to positive antibody reactivity to GTFP-associated herpesvirus antigens, whereas the three controls and four turtles that failed to develop tumors remained negative. Plasma samples from 104 free-ranging green turtles from two Florida (USA) coastal feeding grounds with different GTFP prevalences were tested by ELISA for antibodies to L. learedi adult antigens; and there was no statistically significant association between antibody prevalence and sampling site. When a low optical density cutoff value (0.15) was used to interpret ELISA results, 98% of the turtles from each site were spirorchid antibody-positive and there was no association between antibody reactivity to spirorchids and GTFP status. When a higher negative cutoff value was used, however, a statistically significant association between antibody reactivity to spirorchids and GTFP-free status was found. These results suggest that spirorchids do not have a role in GTFP pathogenesis. All 20 of the tumor-bearing lagoon turtles had antibodies to herpesvirus antigens whereas only two (10%) of the tumor-free reef turtles had detectable anti-herpesvirus reactivity. The strong association between antibody reactivity to herpesvirus antigens and GTFP status in both captive-reared and free-ranging turtles is consistent with the hypothesis that the transmissible agent that causes GTFP is a herpesvirus.

Tetranucleotide markers from the loggerhead sea turtle ( Caretta caretta ) and their cross-amplification in other marine turtle species

The loggerhead sea turtle (Caretta caretta) is a federally threatened species and listed as endangered by the World Conservation Union (IUCN). We describe primers and polymerase chain reaction (PCR) conditions to amplify 11 novel tetranucleotide microsatellite loci from the loggerhead sea turtle. We tested primers using samples from 22 females that nested at Melbourne Beach, Florida (USA). Primer pairs yielded an average of 11.2 alleles per locus (range of 4–24), an average observed heterozygosity of 0.83 (range 0.59–0.96), and an average polymorphic information content of 0.80 (range 0.62–0.94). We also demonstrate the utility of these primers, in addition to primers for 15 loci previously described, for amplifying microsatellite loci in four additional species representing the two extant marine turtle families: olive ridley (Lepidochelys olivacea), hawksbill (Eretmochelys imbricata), green turtle (Chelonia mydas), and leatherback (Dermochelys coriacea).

Modeling and mapping isotopic patterns in the Northwest Atlantic derived from loggerhead sea turtles

Stable isotope analysis can be used to infer geospatial linkages of highly migratory species. Identifying foraging grounds of marine organisms from their isotopic signatures is becoming de rigueur as it has been with terrestrial organisms. Sea turtles are being increasingly studied using a combination of satellite telemetry and stable isotope analysis; these studies along with those from other charismatic, highly vagile, and widely distributed species (e.g., tuna, billfish, sharks, dolphins, whales) have the potential to yield large datasets to develop methodologies to decipher migratory pathways in the marine realm. We collected tissue samples (epidermis and red blood cells) for carbon (δ13C) and nitrogen (δ15N) stable isotope analysis from 214 individual loggerheads (Caretta caretta) in the Northwest Atlantic Ocean (NWA). We used discriminant function analysis (DFA) to examine how well δ13C and δ15N classify loggerhead foraging areas. The DFA model was derived from isotopic signatures of 58 loggerheads equipped with satellite tags to identify foraging locations. We assessed model accuracy with the remaining 156 untracked loggerheads that were captured at their foraging locations. The DFA model correctly identified the foraging ground of 93.0% of individuals with a probability greater than 66.7%. The results of the external validation (1) confirm that assignment models based on tracked loggerheads in the NWA are robust and (2) provide the first independent evidence supporting the use of these models for migratory marine organisms. Additionally, we used these data to generate loggerhead-specific δ13C and δ15N isoscapes, the first for a predator in the Atlantic Ocean. We found a latitudinal trend of δ13C values with higher values in the southern region (20–25 °N) and a more complex pattern with δ15N, with intermediate latitudes (30–35 °N) near large coastal estuaries having higher δ15N-enrichment. These results indicate that this method with further refinement may provide a viable, more spatially-explicit option for identifying loggerhead foraging grounds.

Long-Term Trends in Loggerhead (Caretta caretta) Nesting and Reproductive Success at an Important Western Atlantic Rookery

Abstract The Archie Carr National Wildlife Refuge (ACNWR), located along the central east coast of Florida (USA) in the western North Atlantic, hosts one of the largest loggerhead (Caretta caretta) nesting assemblages in the western Hemisphere. Sea turtle nesting activity has been continuously monitored on this beach for > 31 yrs, representing one of the longest sea turtle reproductive data sets in the world. Between 1982 and 2012, an estimated 358,243 nests were deposited on the ACNWR with an estimated annual mean plus 95% confidence interval of 11,556 ± 1,129 nests. This constitutes 25.4% ± 0.8% of the mean annual Florida Index Nesting Beach Survey loggerhead complement. Mean annual clutch size was 113.9 ± 1.4, resulting in a 55.1% ± 4.0% mean annual hatching success rate and a mean emerging success rate of 53.3% ± 3.7%. The only egg-fate that was statistically correlated with emerging success were eggs washed out by erosion. The loss of eggs by erosion was significantly greater during storm and poststorm years, compared with nonstorm years. Among individual first-time nesting females that were measured, mean straight carapace length was 91.2 ± 0.15 cm and mean curved carapace length was 98.2 ± 0.15 cm. These data suggest that the ACNWR supports the greatest loggerhead nest density per kilometer in Florida, underscoring the importance of the ACNWR as one of the most important nesting habitats for loggerhead turtles in the Western Hemisphere.

Use of Baculovirus-Expressed Glycoprotein H in an Enzyme-Linked Immunosorbent Assay Developed To Assess Exposure to Chelonid Fibropapillomatosis-Associated Herpesvirus and Its Relationship to the Prevalence of Fibropapillomatosis in Sea Turtles

ABSTRACT Chelonid fibropapillomatosis-associated herpesvirus (CFPHV) is an alphaherpesvirus believed to cause marine turtle fibropapillomatosis (FP). A serodiagnostic assay was developed for monitoring sea turtle populations for CFPHV exposure. CFPHV glycoprotein H (gH) expressed in recombinant baculovirus was used in an enzyme-linked immunosorbent assay (ELISA) to detect virus-specific 7S turtle antibodies. Using captive-reared green turtles (Chelonia mydas) with no history of virus exposure as “known negatives” and others with experimentally induced FP as “known positives,” the assay had 100% specificity but low sensitivity, as seroconversion was detected in only half of the turtles bearing experimentally induced tumors. Antibodies were detected only in samples collected after cutaneous fibropapillomas appeared, consistent with observations that tumors are significant sites of virion production and antigen expression and the possibility that prolonged/repeated virus shedding may be required for adequate stimulation of 7S antibody responses to gH. Natural routes of infection, however, may produce higher seroconversion rates. High gH antibody seroprevalences (∼80%) were found among wild green turtles in three Florida localities with different FP prevalences, including one site with no history of FP. In addition, all eight loggerhead turtles (Caretta caretta) tested were seropositive despite FP being uncommon in this species. The possibility that CFPHV infection may be common relative to disease suggests roles for environmental and host factors as modulators of disease expression. Alternatively, the possibility of other antigenically similar herpesviruses present in wild populations cannot be excluded, although antibody cross-reactivity with the lung/eye/trachea disease-associated herpesvirus was ruled out in this study.

Detection of Antibodies to a Disease-Associated Herpesvirus of the Green Turtle, Chelonia mydas

ABSTRACT Lung-eye-trachea disease-associated herpesvirus (LETV) is linked with morbidity and mortality in mariculture-reared green turtles, but its prevalence among and impact on wild marine turtle populations is unknown. An enzyme-linked immunosorbent assay (ELISA) was developed for detection of anti-LETV antibodies and could distinguish LETV-exposed green turtles from those with antibodies to fibropapillomatosis-associated herpesvirus (FPHV). Plasma from two captive-reared green turtles immunized with inactivated LETV served as positive controls. Plasma from 42 healthy captive-reared green turtles and plasma from 30 captive-reared green turtles with experimentally induced fibropapillomatosis (FP) and anti-FPHV antibodies had low ELISA values on LETV antigen. A survey of 19 wild green turtles with and 27 without FP (with and without anti-FPHV antibodies, respectively) identified individuals with antibodies to LETV regardless of their FP status. The seroprevalence of LETV infection was 13%. The presence of antibodies to LETV in plasma samples was confirmed by Western blot and immunohistochemical analyses. These results are the first to suggest that wild Florida green turtles are exposed to LETV or to an antigenically closely related herpesvirus(es) other than FPHV and that FPHV and LETV infections are most likely independent events. This is the first ELISA developed to detect antibodies for a specific herpesvirus infection of marine turtles. The specificity of this ELISA for LETV (ability to distinguish LETV from FPHV) makes it valuable for detecting exposure to this specific herpesvirus and enhances our ability to conduct seroepidemiological studies of these disease-associated agents in marine turtles.

Genetic Barcoding of Marine Leeches ( Ozobranchus spp.) from Florida Sea Turtles and Their Divergence in Host Specificity

Ozobranchus margoi and Ozobranchus branchiatus are the only two species of marine turtle leeches (Ozobranchus spp.) known to inhabit the Atlantic coast of the United States and the Gulf of Mexico. In early reports of fibropapillomatosis (FP) in green turtles (Chelonia mydas), O. branchiatus was implicated as a vector in the transmission of Fibropapilloma‐associated turtle herpesvirus (FPTHV). It is imperative that the leech species be identified to elucidate the role Ozobranchus spp. may play in disease transmission. In this study, Ozobranchus branchiatus has been identified for the first time on a loggerhead (Caretta caretta) turtle, and the molecular data for this species is now available for the first time in GenBank. Both species of leeches were also found infecting a single C. mydas. Using morphological taxonomy combined with distance‐ and character‐based genetic sequence analyses, this study has established a DNA barcode for both species of Ozobranchus spp. leech and has shown it can be applied successfully to the identification of leeches at earlier stages of development when morphological taxonomy cannot be employed. The results suggest a different haplotype may exist for O. branchiatus leeches found on C. caretta versus C. mydas. Leech cocoon residue collected from a C. mydas was identified using the new method.

Intra-annual Loggerhead and Green Turtle Spatial Nesting Patterns

Abstract We analyzed a 15-year (1989–2003) dataset of spatial nesting locations for Loggerhead and Green Turtles along a 40.5-km stretch of beach encompassing the Archie Carr National Wildlife Refuge along the Atlantic coast of Florida. To assess whether there are differences in spatial distribution influenced by temporal site-selection cues, we divided each season into quartiles and analyzed the autocorrelative patterns of the nest distributions within each time frame. Fundamentally, intraspecific differences in nest spatial patterns from the beginning to the end of the nesting season were minor. Though the temporal grain of the analyses may not be able to discern affects of fine-scale fluctuations (e.g., high- and low-tide events), these results suggest that environmental variables that change over the nesting season (e.g., ocean temperatures, daylength, and existing human activities) are not significantly influencing where these sea turtles place their nests.