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Author(s): Banerjee, Shreya M; Frey, Amy; Kurle, Carolyn M; Perrault, Justin R; Stewart, Kelly R | Abstract: Understanding species’ mating systems provides important information about their ecology, life history, and behavior. Direct observations of mating behaviors can be challenging, but molecular techniques can reveal information about mating systems and paternal identity in difficult-to-observe species such as sea turtles. Genetic markers can be used to assess the paternity of a clutch and to assign hatchlings to a father. Leatherback turtles Dermochelys coriacea sometimes mate with multiple individuals, resulting in clutches with mixed paternity; however, the effects of multiple paternity on hatchling quality are unclear. Leatherback hatchlings at Sandy Point National Wildlife Refuge, St. Croix, US Virgin Islands, exhibit visible variation in individual body size, sometimes within the same clutch. We collected morphometrics and tissue samples from hatchlings across multiple nesting seasons (2009, 2012, 2013, 2015, and 2016) and found that hatchlings exhibited small but statistically significant differences in morphometrics between years. We used maternal and hatchling microsatellite genotypes to reconstruct paternal genotypes, assigning fathers to each hatchling. We found multiple paternity in 5 of 17 clutches analyzed and compared differences in morphometrics between full-siblings with differences between half-siblings. We found no significant differences between morphometrics of hatchlings from the same mother but different fathers. We compared within-clutch variances in morphometrics for clutches with and without multiple paternity and found no significant difference in morphological variation between them. Therefore, we could not attribute differences in hatchling size within a clutch to paternal contribution. Understanding other factors affecting hatchling morphology, and other possible fitness metrics, may reveal insights into the benefits, or lack thereof, of polyandry in sea turtles.

Advances in high-throughput sequencing (HTS) technologies coupled with increased interdisciplinary collaboration are rapidly expanding capacity in the scope and scale of wildlife genetic studies. While existing HTS methods can be directly applied to address some evolutionary and ecological questions, certain research goals necessitate tailoring methods to specific study organisms, such as high-throughput genotyping of the same loci that are comparable over large spatial and temporal scales. These needs are particularly common for studies of highly mobile species of conservation concern like marine turtles, where life history traits, limited financial resources and other constraints require affordable, adaptable methods for HTS genotyping to meet a variety of study goals. Here, we present a versatile marine turtle HTS targeted enrichment platform adapted from the recently developed Rapture (RAD-Capture) method specifically designed to meet these research needs. Our results demonstrate consistent enrichment of targeted regions throughout the genome and discovery of candidate variants in all species examined for use in various conservation genetics applications. Accurate species identification confirmed the ability of our platform to genotype over 1,000 multiplexed samples and identified areas for future methodological improvement such as optimization for low initial concentration samples. Finally, analyses within green turtles supported the ability of this platform to identify informative SNPs for stock structure, population assignment and other applications over a broad geographic range of interest to management. This platform provides an additional tool for marine turtle genetic studies and broadens capacity for future large-scale initiatives such as collaborative global marine turtle genetic databases.

Assessments of fisheries by‐catch on marine megafauna are important for developing effective management strategies. For sea turtles, impacts may be evident in nesting population trends, but generally only after a lag, as turtles are slow to mature. We examined 850 loggerhead turtle (Caretta caretta) by‐catch samples collected over 14 years throughout the western North Atlantic to identify source nesting populations. We identified proportions of turtles from 19 distinct management units (MUs), using mitochondrial DNA and mixed stock analysis, considering population size. We found a significant split in the distribution of small versus large loggerheads to the north and south of Cape Hatteras, North Carolina (latitude 35°N). North of Cape Hatteras, large turtles came from south‐east Florida (SEFL; 44% ± 15%) and the northern United States (33% ± 16%) MUs, while south of Cape Hatteras, the major contributors were Central East Florida (52% ± 20%) and south‐east Florida (SEFL; 41% ± 20%). Small turtles in the north came from Central East Florida (64% ± 14%), and in the south, south‐east Florida had the highest representation (56% ± 25%). There was little contribution from Mediterranean and Cape Verde populations, indicating low risk from western North Atlantic by‐catch to MUs in the Mediterranean. Understanding where threats may impact source populations is important, particularly for larger size turtles, as these individuals have the highest reproductive value. These findings allow more informed threat assessments to be conducted for loggerhead turtles considering spatial and demographic structure.

Suicide bereavement is a newly developing field of research in the UK, yet over 6,600 people take their own lives here every year. This heuristic inquiry aims to explore the lived experiences of tr...

Genetic analyses of nuclear DNA (e.g., microsatellites) are a primary tool for investigating mating systems in reptiles, particularly marine turtles. Whereas studies over the past two decades have demonstrated that polyandry (i.e., females mating with multiple males) is common in marine turtles, polygyny (i.e., males mating with multiple females) has rarely been reported. In this study we investigated the mating structure of Critically Endangered hawksbill turtles (Eretmochelys imbricata) at Bahia de Jiquilisco in El Salvador, one of the largest rookeries in the eastern Pacific Ocean. We collected genetic samples from 34 nesting females and hatchlings from 41 clutches during the 2015 nesting season, including one nest from each of 27 females and two nests from seven additional females. Using six highly polymorphic microsatellite loci, we reconstructed the paternal genotypes for 22 known male turtles and discovered that seven (31.8%) sired nests from multiple females, which represents the highest polygyny level reported to date for marine turtles and suggests that this is a common mating structure for this population. We also detected multiple paternity in four (11.8%) clutches from the 34 females analyzed, confirming polyandrous mating strategies are also employed. The high level of polygyny we documented suggests there may be a limited number of sexually mature males at Bahia de Jiquilisco; a scenario supported by multiple lines of empirical evidence. Our findings highlight key management uncertainties, including whether polygynous mating strategies can compensate for potential ongoing feminization and the low number of adult males found for this and possibly other marine turtle populations.

Although the visual and geomagnetic orientation cues used by sea turtle hatchlings during sea-finding have been well studied, the potential for auditory stimuli to act as an orientation cue has not been explored. We investigated the response of sea turtle hatchlings to natural and anthropogenic noises present on their nesting beaches during sea-finding. The responses of hatchling leatherback sea turtles, Dermochelys coriacea, collected from the Sandy Point National Wildlife Refuge, St. Croix, were measured in the presence of aerial acoustic sounds within hatchlings’ hearing range of 50 to 1600 Hz. The highest sound energy produced by beach waves occurs at frequencies 50–1000 Hz, which overlaps with the most sensitive hearing range of hatchling leatherbacks (50–400 Hz). Natural beach wave sounds, which have highest sound energy at frequencies of 50–1000 Hz, may be masked by human conversations (85–650 Hz) and vehicle traffic noise (60–8000 Hz). In the presence of three stimuli, a) beach wave sounds (72.0 dB re: 20 μPa), b) human conversation (72.4 dB re: 20 μPa), and c) vehicle traffic noise (71.1 dB re: 20 μPa), hatchlings exhibited no phonotaxic response (wave sounds: mean angle = 152.1°, p = 0.645; human conversation: mean angle = 67.4°, p = 0.554; traffic noise: mean angle = 125.7°, p = 0.887). These results may be due to the hatchlings being unable to localize sounds in the experimental arena. Visual and auditory cues may also converge to affect sea-finding orientation. Future studies should focus on the localization ability of sea turtles and on the role that sound may play in orientation when combined with other sensory and environmental cues.

Advances in high-throughput sequencing (HTS) technologies coupled with increased interdisciplinary collaboration is rapidly expanding capacity in the scope and scale of wildlife genetic studies. While existing HTS methods can be directly applied to address some evolutionary and ecological questions, certain research goals necessitate tailoring methods to specific study organisms, such as high-throughput genotyping of the same loci that are comparable over large spatial and temporal scales. These needs are particularly common for studies of highly mobile species of conservation concern like marine turtles, where life history traits, limited financial resources and other constraints require affordable, adaptable methods for HTS genotyping to meet a variety of study goals. Here, we present a versatile marine turtle HTS targeted enrichment platform adapted from the recently developed Rapture (RAD-Capture) method specifically designed to meet these research needs. Our results demonstrate consistent enrichment of targeted regions throughout the genome and discovery of candidate variants in all species examined for use in various conservation genetics applications. Accurate species identification confirmed the ability of our platform to genotype over 1,000 multiplexed samples, and identified areas for future methodological improvement such as optimization for low initial concentration samples. Finally, analyses within green turtles supported the ability of this platform to identify informative SNPs for stock structure, population assignment and other applications over a broad geographic range of interest to management. This platform provides an additional tool for marine turtle genetic studies and broadens capacity for future large-scale initiatives such as collaborative global marine turtle genetic databases.

Understanding population dynamics in broadly distributed marine species with cryptic life history stages is challenging. Information on the population dynamics of sea turtles tends to be biased toward females, due to their accessibility for study on nesting beaches. Males are encountered only at sea; there is little information about their migratory routes, residence areas, foraging zones, and population boundaries. In particular, male leatherbacks (Dermochelys coriacea) are quite elusive; little is known about adult and juvenile male distribution or behavior. The at-sea distribution of male turtles from different breeding populations is not known. Here, 122 captured or stranded male leatherback turtles from the USA, Turkey, France, and Canada (collected 1997–2012) were assigned to one of nine Atlantic basin populations using genetic analysis with microsatellite DNA markers. We found that all turtles originated from western Atlantic nesting beaches (Trinidad 55%, French Guiana 31%, and Costa Rica 14%). Although genetic data for other Atlantic nesting populations were represented in the assignment analysis (St. Croix, Brazil, Florida, and Africa (west and south), none of the male leatherbacks included in this study were shown to originate from these populations. This was an unexpected result based on estimated source population sizes. One stranded turtle from Turkey was assigned to French Guiana, while others that were stranded in France were from Trinidad or French Guiana breeding populations. For 12 male leatherbacks in our dataset, natal origins determined from the genetic assignment tests were compared to published satellite and flipper tag information to provide evidence of natal homing for male leatherbacks, which corroborated our genetic findings. Our focused study on male leatherback natal origins provides information not previously known for this cryptic, but essential component of the breeding population. This method should provide a guideline for future studies, with the ultimate goal of improving management and conservation strategies for threatened and endangered species by taking the male component of the breeding population into account.

Despite facing serious threats of extinction in the Eastern Pacific, the leatherback turtle ( Dermochelys coriacea ) appears to be thriving in the Atlantic basin based on increasing nest counts at several rookeries. In particular, Florida’s nest numbers have been increasing by 10.2% per year since standardized counts began in 1979. The US Recovery Plan for leatherbacks calls for vital rates and population parameters to be determined for the three leatherback rookeries under US jurisdiction: St. Croix (USVI), Puerto Rico, and the east coast of Florida. Based on mark-recapture data gathered over eleven years, we determined important population parameters for nesting female leatherbacks at Juno Beach, one of the most densely nested beaches in Florida. Average annual survival was 88.9%. The average female nesting population size for Juno Beach is estimated at 100 ± 41 individuals each season; statewide we expect the estimate to be higher. The average remigration interval was 2.7 ± 1.0 years. In addition, we report observed clutch frequency (2.1 ± 1.4 clutches/year), estimated clutch frequency (4.4 ± 1.1 nests/year), and observed internesting period (10.2 ± 1.3 days between nests). The probability of observing an individual female at least once during the season was 73.0%, likely due to variable site fidelity, even though sea turtles do exhibit natal homing. Using opportunistic observations at additional beaches, we found that 72 females observed nesting within the Juno Beach study area were also observed nesting outside the study area. Thirty-three individuals laid clutches both inside and outside the survey area within a single season; these nests were separated by as much as 463.5 km. Although the population in Florida is relatively small compared to other rookeries throughout the Western Atlantic, it is increasing at such a rapid pace that it has the potential to become more important regionally, thereby contributing to the abundance of leatherbacks in the Atlantic.

Leatherback sea turtles (Dermochelys coriacea) are listed as Vulnerable globally by the International Union for Conservation of Nature. To preserve this species, we must protect them during several life stages, including protecting hatchlings emerging from nests. We studied the timing of emergence of leatherback hatchlings at Sandy Point National Wildlife Refuge, an area created for the protection of this species. We surveyed 524 nests in 2010–2014 and evaluated the time that hatchlings began emerging from the surface of the sand, the time that the last hatchling in the group finished emerging from the sand, the incubation duration of each nest (number of days between deposition and first emergence), and the number of hatchlings in the initial pulse of emergence. In situ clutches (those occurring naturally and not manipulated by location) had an incubation period of 61.4 ± 21.1 d (mean ± SD; n = 332) and emerged from 1825 to 1906 hrs. The incubation duration varied significantly from relocated clutches (those that had been moved at deposition because of erosion threats), had an incubation period of 60.0 ± 2.7 days (n = 192), and emerged from 1827 to 1903 hrs (on average, for all years combined; p < 0.0001). These nests yielded 36.6 ± 21.1 first-emergence hatchlings from in situ clutches, which was different from relocated clutches (14.9 ± 13.6 first-emergence; p < 0.0001). Other studies have found that hard-shelled sea turtles all begin emerging once the sun sets and continue emerging throughout the night. In contrast, we found that leatherback sea turtle hatchlings emerge earlier in the evening, at least at Sandy Point. During the seasons sampled, emergence often began before sunset (between 1834 and 1859 hrs) and hatchlings usually did not continue to emerge through the night. To allocate resources most effectively, beach management programs at Sandy Point National Wildlife Refuge should be maximized between 1700 and 2000 hrs, the period when most hatchlings are emerging and require the most protection from predators. This work provides important information about the timing of leatherback emergence and may be applicable to other leatherback nesting beaches where predation may be a problem. These findings can help ensure that nest protection programs maintain efficiency and protect hatchlings during their vulnerable crawl to the sea.

Investigating migratory connectivity between breeding and foraging areas is critical to effective management and conservation of highly mobile marine taxa, particularly threatened, endangered, or economically important species that cross through regional, national and international boundaries. The leatherback turtle (Dermochelys coriacea, Vandelli 1761) is one such transboundary species that spends time at breeding areas at low latitudes in the northwest Atlantic during spring and summer. From there, they migrate widely throughout the North Atlantic, but many show fidelity to one region off eastern Canada, where critical foraging habitat has been proposed. Our goal was to identify nesting beach origins for turtles foraging here. Using genetics, we identified natal beaches for 288 turtles that were live-captured off the coast of Nova Scotia, Canada. Turtles were sampled (skin or blood) and genotyped using 17 polymorphic microsatellite markers. Results from three assignment testing programs (ONCOR, GeneClass2 and Structure) were compared. Our nesting population reference data set included 1417 individuals from nine Atlantic nesting assemblages. A supplementary data set for 83 foraging turtles traced to nesting beaches using flipper tags and/or PIT tags (n = 72), or inferred from satellite telemetry (n = 11), enabled ground-truthing of the assignments. We first assigned turtles using only genetic information and then used the supplementary recapture information to verify assignments. ONCOR performed best, assigning 64 of the 83 recaptured turtles to natal beaches (77·1%). Turtles assigned to Trinidad (164), French Guiana (72), Costa Rica (44), St. Croix (7), and Florida (1) reflect the relative size of those nesting populations, although none of the turtles were assigned to four other potential source nesting assemblages. Our results demonstrate the utility of genetic approaches for determining source populations of foraging marine animals and include the first identification of natal rookeries of male leatherbacks, identified through satellite telemetry and verified with genetics. This work highlights the importance of long-term monitoring and tagging programmes in nesting and high-use foraging areas. Moreover, it provides a scientific basis for evaluating stock-specific effects of fisheries on migratory marine species, thus identifying where coordinated international recovery efforts may be most effective.

This study presents a comprehensive genetic analysis of stock structure for leatherback turtles (Dermochelys coriacea), combining 17 microsatellite loci and 763 bp of the mtDNA control region. Recently discovered eastern Atlantic nesting populations of this critically endangered species were absent in a previous survey that found little ocean-wide mtDNA variation. We added rookeries in West Africa and Brazil and generated longer sequences for previously analyzed samples. A total of 1,417 individuals were sampled from nine nesting sites in the Atlantic and SW Indian Ocean. We detected additional mtDNA variation with the longer sequences, identifying ten polymorphic sites that resolved a total of ten haplotypes, including three new variants of haplotypes previously described by shorter sequences. Population differentiation was substantial between all but two adjacent rookery pairs, and F ST values ranged from 0.034 to 0.676 and 0.004 to 0.205 for mtDNA and microsatellite data respectively, suggesting that male-mediated gene flow is not as widespread as previously assumed. We detected weak (F ST = 0.008 and 0.006) but significant differentiation with microsatellites between the two population pairs that were indistinguishable with mtDNA data. POWSIM analysis showed that our mtDNA marker had very low statistical power to detect weak structure (F ST

Fisheries that operate at large spatial scales and with high intensity have the potential to impact highly migratory species, and it is important to characterize threats to specific breeding populations of these species. We used many‐to‐many mixed‐stock analysis (MSA) (n = 408) and microsatellite assignment testing (n = 397) to determine source populations for leatherback turtles (Dermochelys coriacea) caught as bycatch in the U.S. pelagic longline fishery from 2002 to 2012 in the western North Atlantic. Within the United States, we had bycatch samples from the majority of statistical fishing areas: Gulf of Mexico (GOM), Northeast Distant (NED), Caribbean (CAR), Florida East Coast (FEC), Mid Atlantic Bight (MAB), Northeast Coastal (NEC), South Atlantic Bight (SAB) and Sargasso (SAR). We determined the proportions of turtles from each of nine nesting stocks in the Atlantic in each of the sampled areas. These nesting stocks included Brazil, Costa Rica, Florida, Trinidad, French Guiana, St. Croix, Ghana, Gabon, and South Africa. The MSA revealed that the NED had a lower relative proportion of turtles from Costa Rica than other areas and that the GOM had the highest relative proportion of turtles from Costa Rica. No turtles were assigned to the African rookeries, lending further evidence that turtles from that region forage elsewhere and therefore may not be affected by western North Atlantic fisheries. This work contributes to the ongoing assessment of threats to leatherback turtles in the Regional Management Unit (RMU) of the western North Atlantic, and draws attention to the disproportionate number of turtles from Costa Rica being caught in the Gulf of Mexico; Costa Rica is one of the only populations in the northern Atlantic that is not experiencing significant increases in nest numbers. This approach should be useful in determining population‐specific threats to other highly migratory protected species that may depend on segregated foraging areas either within or among species.

When animals are difficult to observe while breeding, insights into the mating system may be gained by using molecular techniques. Patterns of extra-pair copulation, multiple paternity and parental genotype analysis may elucidate population characteristics that help improve knowledge of life history while informing management decisions. During the course of a long-term study of leatherback turtles, we assessed the level of multiple paternity in successive clutches for 12 known females nesting at Sandy Point National Wildlife Refuge (St. Croix, U.S. Virgin Islands). We used seven polymorphic microsatellite markers to genotype the females and 1,019 hatchlings representing 38 nests (3–4 clutches from each female). Using deductive genotype reconstruction and GERUD1.0, we identified the 12 mothers and 17 different fathers that were responsible for 38 nests. We found that seven females (58.3%) showed no evidence of multiple paternity in their clutches, while five females (41.7%) had mated with two males each. There was evidence of two fathers (polyandry) in successive clutches for these five females. Multiple fathers didn’t contribute to clutches equally. For clutches laid by an individual female, the primary father was responsible for 53.7 to 85.9% of the hatchlings. We demonstrate the feasibility of using male genotype reconstruction to characterize the male component of this breeding population and to assess operational sex ratios for breeding sea turtles.

Understanding population status for endangered species is critical to developing and evaluating recovery plans mandated by the Endangered Species Act. For sea turtles, changes in abundance are difficult to detect because most life stages occur in the water. Currently, nest counts are the most reliable way of assessing trends. We determined the rate of growth for leatherback turtle (Dermochelys coriacea) nest numbers in Florida (USA) using a multilevel Poisson regression. We modeled nest counts from 68 beaches over 30 years and, using beach-level covariates (including latitude), we allowed for partial pooling of information between neighboring beaches. This modeling approach is ideal for nest count data because it recognizes the hierarchical structure of the data while incorporating variables related to survey effort. Nesting has increased at all 68 beaches in Florida, with trends ranging from 3.1% to 16.3% per year. Overall, across the state, the number of nests has been increasing by 10.2% per year since 1979. Despite being a small population (probably

We collected tissue samples from 41 nesting hawksbill turtles (Eretmochelys imbricata) at Sandy Point National Wildlife Refuge, St. Croix, US Virgin Islands, to characterize the genetic structure of this rookery in terms of mitochondrial DNA; we compared haplotype frequencies from this rookery to those from Buck Island, another hawksbill nesting beach on St. Croix. Pairwise FST comparisons showed that Sandy Point was demographically distinct from Buck Island (FST = 0.501, p < 0.001), a finding reinforced by significantly different haplotype frequencies (χ2 = 51.76, p < 0.001) and a lack of interchange of nesting females between both sites based on mark–recapture data. Our results support the delineation of the nesting populations at Sandy Point and Buck Island into separate units for the purposes of management.

Although regulated fishing effort is relatively well documented for fisheries in developed states, developing countries are dominated by artisanal fisheries that are characterized by large numbers of small boats, fishing in dispersed and remote locations. These factors make quantifying artisanal fishing effort difficult. In this study, we examined the distribution and density of fishing effort across a region dominated by coastal, artisanal fisheries: the wider Caribbean. We used generalized linear regression models to predict missing data needed to compute fishing effort metrics and to explain variance in average boat length of a fishery and the number of small-scale boats in a given country. Clear intra-regional differences between mainland and island fisheries, and between northern and southern Caribbean fisheries, are evident in the results. To map artisanal fisheries based on the minimal data available, we created a free, automated Fishing Effort Envelope Tool (FEET). Through the use of this tool, we mapped all fisheries in the Caribbean to the extent possible given current data. Further, this mapping process also allowed us to identify hotspots of high density coastal fishing and data gaps that may mask areas of even higher fishing pressure. The potential ecological consequences of the scale of artisanal fishing are profound, and have greater implications for developing regions worldwide.

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 leathe...

Recent research on ocean health has found large predator abundance to be a key element of ocean condition. Fisheries can impact large predator abundance directly through targeted capture and indirectly through incidental capture of nontarget species or bycatch. However, measures of the global nature of bycatch are lacking for air-breathing megafauna. We fill this knowledge gap and present a synoptic global assessment of the distribution and intensity of bycatch of seabirds, marine mammals, and sea turtles based on empirical data from the three most commonly used types of fishing gears worldwide. We identify taxa-specific hotspots of bycatch intensity and find evidence of cumulative impacts across fishing fleets and gears. This global map of bycatch illustrates where data are particularly scarce--in coastal and small-scale fisheries and ocean regions that support developed industrial fisheries and millions of small-scale fishers--and identifies fishing areas where, given the evidence of cumulative hotspots across gear and taxa, traditional species or gear-specific bycatch management and mitigation efforts may be necessary but not sufficient. Given the global distribution of bycatch and the mitigation success achieved by some fleets, the reduction of air-breathing megafauna bycatch is both an urgent and achievable conservation priority.

For vertebrates with temperature-dependent sex determination, primary (or hatchling) sex ratios are often skewed, an issue of particular relevance to concerns over effects of climate change on populations. However, the ratio of breeding males to females, or the operational sex ratio (OSR), is important to understand because it has consequences for population demographics and determines the capacity of a species to persist. The OSR also affects mating behaviors and mate choice, depending on the more abundant sex. For sea turtles, hatchling and juvenile sex ratios are generally female-biased, and with warming nesting beach temperatures, there is concern that populations may become feminized. Our purpose was to evaluate the breeding sex ratio for leatherback turtles at a nesting beach in St. Croix, USVI. In 2010, we sampled nesting females and later sampled their hatchlings as they emerged from nests. Total genomic DNA was extracted and all individuals were genotyped using 6 polymorphic microsatellite markers. We genotyped 662 hatchlings from 58 females, matching 55 females conclusively to their nests. Of the 55, 42 females mated with one male each, 9 mated with 2 males each and 4 mated with at least 3 males each, for a multiple paternity rate of 23.6%. Using GERUD1.0, we reconstructed parental genotypes, identifying 47 different males and 46 females for an estimated breeding sex ratio of 1.02 males for every female. Thus we demonstrate that there are as many actively breeding males as females in this population. Concerns about female-biased adult sex ratios may be premature, and mate choice or competition may play more of a role in sea turtle reproduction than previously thought. We recommend monitoring breeding sex ratios in the future to allow the integration of this demographic parameter in population models.

Previous genetic studies have demonstrated that natal homing shapes the stock structure of marine turtle nesting populations. However, widespread sharing of common haplotypes based on short segments of the mitochondrial control region often limits resolution of the demographic connectivity of populations. Recent studies employing longer control region sequences to resolve haplotype sharing have focused on regional assessments of genetic structure and phylogeography. Here we synthesize available control region sequences for loggerhead turtles from the Mediterranean Sea, Atlantic, and western Indian Ocean basins. These data represent six of the nine globally significant regional management units (RMUs) for the species and include novel sequence data from Brazil, Cape Verde, South Africa and Oman. Genetic tests of differentiation among 42 rookeries represented by short sequences (380 bp haplotypes from 3,486 samples) and 40 rookeries represented by long sequences (~800 bp haplotypes from 3,434 samples) supported the distinction of the six RMUs analyzed as well as recognition of at least 18 demographically independent management units (MUs) with respect to female natal homing. A total of 59 haplotypes were resolved. These haplotypes belonged to two highly divergent global lineages, with haplogroup I represented primarily by CC-A1, CC-A4, and CC-A11 variants and haplogroup II represented by CC-A2 and derived variants. Geographic distribution patterns of haplogroup II haplotypes and the nested position of CC-A11.6 from Oman among the Atlantic haplotypes invoke recent colonization of the Indian Ocean from the Atlantic for both global lineages. The haplotypes we confirmed for western Indian Ocean RMUs allow reinterpretation of previous mixed stock analysis and further suggest that contemporary migratory connectivity between the Indian and Atlantic Oceans occurs on a broader scale than previously hypothesized. This study represents a valuable model for conducting comprehensive international cooperative data management and research in marine ecology.

The authors compared blood plasma concentrations of 13 perfluoroalkyl contaminants (PFCs) in five sea turtle species with differing trophic levels. Wild sea turtles were blood sampled from the southeastern region of the United States, and plasma was analyzed using liquid chromatography tandem mass spectrometry. Mean concentrations of perfluorooctane sulfonate (PFOS), the predominant PFC, increased with trophic level from herbivorous greens (2.41 ng/g), jellyfish-eating leatherbacks (3.95 ng/g), omnivorous loggerheads (6.47 ng/g), to crab-eating Kemp's ridleys (15.7 ng/g). However, spongivorous hawksbills had surprisingly high concentrations of PFOS (11.9 ng/g) and other PFCs based on their trophic level. These baseline concentrations of biomagnifying PFCs demonstrate interesting species and geographical differences. The measured PFOS concentrations were compared with concentrations known to cause toxic effects in laboratory animals, and estimated margins of safety (EMOS) were calculated. Small EMOS (

To assess threats to endangered species, it is critical to establish baselines for contaminant concentrations that may have detrimental consequences to individuals or populations. We measured contaminants in blubber and fat from dead leatherback turtles and established baselines in blood and eggs in nesting turtles. In fat, blubber, blood and eggs, the predominant PCBs were 153 + 132, 187 + 182, 138 + 163, 118, and 180 + 193. Total PCBs, 4,4′-DDE, total PBDEs and total chlordanes were significantly and positively correlated between blood and eggs, suggesting maternal transfer. Significant positive relationships also existed between fat and blubber in stranded leatherbacks. Less lipophilic PCBs appeared to more readily transfer from females to their eggs. PBDE profiles in the four tissues were similar to other wildlife populations but different from some turtle studies. Concentrations were lower than those shown to have acute toxic effects in other aquatic reptiles, but may have sub-lethal effects on hatchling body condition and health.

Biodiverse coastal zones are often areas of intense fishing pressure due to the high relative density of fishing capacity in these nearshore regions. Although overcapacity is one of the central challenges to fisheries sustainability in coastal zones, accurate estimates of fishing pressure in coastal zones are limited, hampering the assessment of the direct and collateral impacts (e.g., habitat degradation, bycatch) of fishing. We compiled a comprehensive database of fishing effort metrics and the corresponding spatial limits of fisheries and used a spatial analysis program (FEET) to map fishing effort density (measured as boat-meters per km2) in the coastal zones of six ocean regions. We also considered the utility of a number of socioeconomic variables as indicators of fishing pressure at the national level; fishing density increased as a function of population size and decreased as a function of coastline length. Our mapping exercise points to intra and interregional ‘hotspots’ of coastal fishing pressure. The significant and intuitive relationships we found between fishing density and population size and coastline length may help with coarse regional characterizations of fishing pressure. However, spatially-delimited fishing effort data are needed to accurately map fishing hotspots, i.e., areas of intense fishing activity. We suggest that estimates of fishing effort, not just target catch or yield, serve as a necessary measure of fishing activity, which is a key link to evaluating sustainability and environmental impacts of coastal fisheries.

The advantages and disadvantages of the immunofluorescence and immunoperoxidase techniques and an introductory description of the major variations of the basic immunoperoxidase method are discussed. The ability to identify intermediate filaments, T and B lymphocyte cell markers, S-100 proteins, myelin basic protein, and carcinoem-bryonic antigen in tissue sections can assist the dermatopathologist in classifying undifferentiated or histologically similar-appearing tumors. A systematic approach to the diagnosis of a majority of these tumors with the use of immunoenzyme staining is provided.