Michael P. Jensen

Genetic Stock Structure of Green Turtle (Chelonia mydas) Nesting Populations across the Pacific Islands

Abstract: More than two decades have passed since the first studies documenting genetic population structure of green turtles (Chelonia mydas) were published. Since then many more have followed and characterization of the genetic structure of green turtle rookeries now covers most of the global distribution of the species, benefitting conservation of this threatened species worldwide. However, important data gaps still exist across a large part of the western and central Pacific Ocean (WCPO). This large area is made up of hundreds of scattered islands and atolls of Micronesia, Melanesia, and Polynesia, most of which are remote and difficult to access. In this study, we assessed stock structure of green turtles throughout the WCPO using mitochondrial (mt) DNA from 805 turtles sampled across 25 nesting locations. We examined whether sequencing longer fragments (770 bp) of the control region increases resolution of stock structure and used genetic analysis to evaluate level of demographic connectivity among island nesting populations in the WCPO. We identified a total of 25 haplotypes characterized by polymorphism within the 770 bp sequences, including five new variants of haplotypes that were indistinguishable with shorter 384 bp reads from previous studies. Stock structure analysis indicated that rookeries separated by more than 1,000 km were significantly differentiated from each other, but neighboring rookeries within 500 km showed no genetic differentiation. Results presented in this paper establish that sequencing of longer fragments (770 bp) of the control region does in some cases increase resolution and that there are at least seven independent stocks in the region.

Migration of green turtles (Chelonia mydas) from Australasian feeding grounds inferred from genetic analyses

Coastal seagrass habitats in tropical and subtropical regions support aggregations of resident green turtles (Chelonia mydas) from several genetically distinct breeding populations. Migration of individuals to their respective dispersed breeding sites provides a complex pattern of migratory connectivity among nesting and feeding habitats of this species. An understanding of this pattern is important in regions where the persistence of populations is under threat from anthropogenic impacts. The present study uses mitochondrial DNA and mixed-stock analyses to assess the connectivity among seven feeding grounds across the north Australian coast and adjacent areas and 17 genetically distinct breeding populations from the Indo-Pacific region. It was hypothesised that large and geographically proximate breeding populations would dominate at nearby feeding grounds. As expected, each sampled feeding area appears to support multiple breeding populations, with two aggregations dominated by a local breeding population. Geographic distance between breeding and feeding habitat strongly influenced whether a breeding population contributed to a feeding ground (wi = 0.654); however, neither distance nor size of a breeding population was a good predictor of the extent of their contribution. The differential proportional contributions suggest the impact of anthropogenic mortality at feeding grounds should be assessed on a case-by-case basis.

Population structure and phylogeography reveal pathways of colonization by a migratory marine reptile (Chelonia mydas) in the central and eastern Pacific.

Climate, behavior, ecology, and oceanography shape patterns of biodiversity in marine faunas in the absence of obvious geographic barriers. Marine turtles are an example of highly migratory creatures with deep evolutionary lineages and complex life histories that span both terrestrial and marine environments. Previous studies have focused on the deep isolation of evolutionary lineages (>3 mya) through vicariance; however, little attention has been given to the pathways of colonization of the eastern Pacific and the processes that have shaped diversity within the most recent evolutionary time. We sequenced 770 bp of the mtDNA control region to examine the stock structure and phylogeography of 545 green turtles from eight different rookeries in the central and eastern Pacific. We found significant differentiation between the geographically separated nesting populations and identified five distinct stocks (FST = 0.08–0.44, P < 0.005). Central and eastern Pacific Chelonia mydas form a monophyletic group containing 3 subclades, with Hawaii more closely related to the eastern Pacific than western Pacific populations. The split between sampled central/eastern and western Pacific haplotypes was estimated at around 0.34 mya, suggesting that the Pacific region west of Hawaii has been a more formidable barrier to gene flow in C. mydas than the East Pacific Barrier. Our results suggest that the eastern Pacific was colonized from the western Pacific via the Central North Pacific and that the Revillagigedos Islands provided a stepping-stone for radiation of green turtles from the Hawaiian Archipelago to the eastern Pacific. Our results fit with a broader paradigm that has been described for marine biodiversity, where oceanic islands, such as Hawaii and Revillagigedo, rather than being peripheral evolutionary “graveyards”, serve as sources and recipients of diversity and provide a mechanism for further radiation.

Phylogeography, Genetic Diversity, and Management Units of Hawksbill Turtles in the Indo-Pacific

Hawksbill turtle (Eretmochelys imbricata) populations have experienced global decline because of a history of intense commercial exploitation for shell and stuffed taxidermied whole animals, and harvest for eggs and meat. Improved understanding of genetic diversity and phylogeography is needed to aid conservation. In this study, we analyzed the most geographically comprehensive sample of hawksbill turtles from the Indo-Pacific Ocean, sequencing 766 bp of the mitochondrial control region from 13 locations (plus Aldabra, n = 4) spanning over 13500 km. Our analysis of 492 samples revealed 52 haplotypes distributed in 5 divergent clades. Diversification times differed between the Indo-Pacific and Atlantic lineages and appear to be related to the sea-level changes that occurred during the Last Glacial Maximum. We found signals of demographic expansion only for turtles from the Persian Gulf region, which can be tied to a more recent colonization event. Our analyses revealed evidence of transoceanic migration, including connections between feeding grounds from the Atlantic Ocean and Indo-Pacific rookeries. Hawksbill turtles appear to have a complex pattern of phylogeography, showing a weak isolation by distance and evidence of multiple colonization events. Our novel dataset will allow mixed-stock analyses of hawksbill turtle feeding grounds in the Indo-Pacific by providing baseline data needed for conservation efforts in the region. Eight management units are proposed in our study for the Indo-Pacific region that can be incorporated in conservation plans of this critically endangered species.

Hawksbill turtle terra incognita: conservation genetics of eastern Pacific rookeries

Abstract Prior to 2008 and the discovery of several important hawksbill turtle (Eretmochelys imbricata) nesting colonies in the EP (Eastern Pacific), the species was considered virtually absent from the region. Research since that time has yielded new insights into EP hawksbills, salient among them being the use of mangrove estuaries for nesting. These recent revelations have raised interest in the genetic characterization of hawksbills in the EP, studies of which have remained lacking to date. Between 2008 and 2014, we collected tissue samples from 269 nesting hawksbills at nine rookeries across the EP and used mitochondrial DNA sequences (766 bp) to generate the first genetic characterization of rookeries in the region. Our results inform genetic diversity, population differentiation, and phylogeography of the species. Hawksbills in the EP demonstrate low genetic diversity: We identified a total of only seven haplotypes across the region, including five new and two previously identified nesting haplotypes (pooled frequencies of 58.4% and 41.6%, respectively), the former only evident in Central American rookeries. Despite low genetic diversity, we found strong stock structure between the four principal rookeries, suggesting the existence of multiple populations and warranting their recognition as distinct management units. Furthermore, haplotypes EiIP106 and EiIP108 are unique to hawksbills that nest in mangrove estuaries, a behavior found only in hawksbills along Pacific Central America. The detected genetic differentiation supports the existence of a novel mangrove estuary “reproductive ecotype” that may warrant additional conservation attention. From a phylogeographic perspective, our research indicates hawksbills colonized the EP via the Indo‐Pacific, and do not represent relict populations isolated from the Atlantic by the rising of the Panama Isthmus. Low overall genetic diversity in the EP is likely the combined result of few rookeries, extremely small reproductive populations and evolutionarily recent colonization events. Additional research with larger sample sizes and variable markers will help further genetic understanding of hawksbill turtles in the EP.

Mixed stock analysis of a resident green turtle, Chelonia mydas, population in New Caledonia links rookeries in the South Pacific

Abstract Context. Migratory species are known to pose a challenge for conservation because it is essential to understand their complex life history in order to implement efficient conservation actions. Aims. In New Caledonia, large seagrass habitats in the Grand Lagon Sud (GLS) are home to resident green turtles (Chelonia mydas) of unknown origins. To assess the stock composition in the GLS, 164 foraging turtles were sampled for genetic analysis of ∼770 base pairs of the mitochondrial DNA (mtDNA) control region. Methods. Foraging turtles ranging in size from 48.0 to 108.4 cm curved carapace length were captured at five different sites within the GLS between September 2012 and December 2013. To provide baseline data for mixed stock analysis, published data from rookeries were used in addition to 105 samples collected at rookeries in the d’Entrecasteaux Islands and Chesterfield Islands in New Caledonia and at Malekula Island in Vanuatu. Exact tests of population differentiation and pairwise FST estimates were used to test for differences in mtDNA haplotype frequencies. Key results. These analyses indicated that rookeries in the d’Entrecasteaux Islands and Vanuatu form unique management units and that the Chesterfield Islands rookeries are linked to the Coral Sea management unit. Mixed stock analysis indicated the highest proportion (mean = 0.63) of foraging turtles originate from the d’Entrecasteaux stock. Conclusions. The larger contribution is estimated to be from a large rookery from New Caledonia, but smaller contributions are suggested from other rookeries in the South Pacific. Implications. Marine conservation policies in New Caledonia need to consider the links between the foraging and nesting populations of C. mydas in New Caledonia and other rookeries and foraging grounds in the Coral Sea.

Does polyandry really pay off? The effects of multiple mating and number of fathers on morphological traits and survival in clutches of nesting green turtles at Tortuguero

Despite the long debate of whether or not multiple mating benefits the offspring, studies still show contradictory results. Multiple mating takes time and energy. Thus, if females fertilize their eggs with a single mating, why to mate more than once? We investigated and inferred paternal identity and number of sires in 12 clutches (240 hatchlings) of green turtles (Chelonia mydas) nests at Tortuguero, Costa Rica. Paternal alleles were inferred through comparison of maternal and hatchling genotypes, and indicated multiple paternity in at least 11 of the clutches (92%). The inferred average number of fathers was three (ranging from 1 to 5). Moreover, regression analyses were used to investigate for correlation of inferred clutch paternity with morphological traits of hatchlings fitness (emergence success, length, weight and crawling speed), the size of the mother, and an environmental variable (incubation temperature). We suggest and propose two different comparative approaches for evaluating morphological traits and clutch paternity, in order to infer greater offspring survival. First, clutches coded by the exact number of fathers and second by the exact paternal contribution (fathers who gives greater proportion of the offspring per nest). We found significant differences (P < 0.05) in clutches coded by the exact number of fathers for all morphological traits. A general tendency of higher values in offspring sired by two to three fathers was observed for the length and weight traits. However, emergence success and crawling speed showed different trends which unable us to reach any further conclusion. The second approach analysing the paternal contribution showed no significant difference (P > 0.05) for any of the traits. We conclude that multiple paternity does not provide any extra benefit in the morphological fitness traits or the survival of the offspring, when analysed following the proposed comparative statistical methods.

Advances in the Application of Genetics in Marine Turtle Biology and Conservation

Marine turtles migrate across long distances, exhibit complex life histories, and occupy habitats that are difficult to observe. These factors present substantial challenges to understanding fundamental aspects of their biology or assessing human impacts, many of which are important for the effective conservation of these threatened and endangered species. The early development and application of genetic tools made important contributions to understanding marine turtle population and evolutionary biology, such as providing evidence of regional natal homing by breeding adults, establishing connectivity between rookeries and foraging habitats, and determining phylogeography and broad scale stock structure for most marine turtle species. Recent innovations in molecular technologies, statistical methods, and creative application of genetic tools have significantly built upon this knowledge to address key questions in marine turtle biology and conservation management. Here, we evaluate the latest major advances and potential of marine turtle genetic applications, including improved resolution and large-scale syntheses of population structure, connectivity and phylogeography, estimation of key demographic rates such as age to maturity and operational or breeding sex ratios, insight into reproductive strategies and behavior, and assessment of differential human impacts among populations. We then discuss remaining challenges and emerging capabilities, such as rapid, multiplexed genotyping and investigation of the genomic underpinnings of adaptive variation afforded by high-throughput sequencing technologies.

Natal foraging philopatry in eastern Pacific hawksbill turtles

The complex processes involved with animal migration have long been a subject of biological interest, and broad-scale movement patterns of many marine turtle populations still remain unresolved. While it is widely accepted that once marine turtles reach sexual maturity they home to natal areas for nesting or reproduction, the role of philopatry to natal areas during other life stages has received less scrutiny, despite widespread evidence across the taxa. Here we report on genetic research that indicates that juvenile hawksbill turtles (Eretmochelys imbricata) in the eastern Pacific Ocean use foraging grounds in the region of their natal beaches, a pattern we term natal foraging philopatry. Our findings confirm that traditional views of natal homing solely for reproduction are incomplete and that many marine turtle species exhibit philopatry to natal areas to forage. Our results have important implications for life-history research and conservation of marine turtles and may extend to other wide-ranging marine vertebrates that demonstrate natal philopatry.

Closing the gap: mixed stock analysis of three foraging populations of green turtles (Chelonia mydas) on the Great Barrier Reef

A solid understanding of the spatial ecology of green turtles (Chelonia mydas) is fundamental to their effective conservation. Yet this species, like many marine migratory species, is challenging to monitor and manage because they utilise a variety of habitats that span wide spatio-temporal scales. To further elucidate the connectivity between green turtle rookeries and foraging populations, we sequenced the mtDNA control region of 278 turtles across three foraging sites from the northern Great Barrier Reef (GBR) spanning more than 330 km: Cockle Bay, Green Island and Low Isles. This was performed with a newly developed assay, which targets a longer fragment of mtDNA than previous studies. We used a mixed stock analysis (MSA), which utilises genetic data to estimate the relative proportion of genetically distinct breeding populations found at a given foraging ground. Haplotype and nucleotide diversity was also assessed. A total of 35 haplotypes were identified across all sites, 13 of which had not been found previously in any rookery. The MSA showed that the northern GBR (nGBR), Coral Sea (CS), southern GBR (sGBR) and New Caledonia (NC) stocks supplied the bulk of the turtles at all three sites, with small contributions from other rookeries in the region. Stock contribution shifted gradually from north to south, although sGBR/CS stock dominated at all three sites. The major change in composition occured between Cockle Bay and Low Isles. Our findings, together with other recent studies in this field, show that stock composition shifts with latitude as a natural progression along a coastal gradient. This phenomenon is likely to be the result of ocean currents influencing both post-hatchling dispersal and subsequent juvenile recruitment to diverse coastal foraging sites.