Biochemical Indices and Life Traits of Loggerhead Turtles (Caretta caretta) from Cape Verde Islands

The loggerhead turtle (Caretta caretta) is an endangered marine reptile for whom assessing population health requires knowledge of demographic parameters such as individual growth rate. In Cape Verde, as within several populations, adult female loggerhead sea turtles show a size-related behavioral and trophic dichotomy. While smaller females are associated with oceanic habitats, larger females tend to feed in neritic habitats, which is reflected in their physiological condition and in their offspring. The ratio of RNA/DNA provides a measure of cellular protein synthesis capacity, which varies depending on changes in environmental conditions such as temperature and food availability. The purpose of this study was to evaluate the combined use of morphometric data and biochemical indices as predictors of the physiological condition of the females of distinct sizes and hatchlings during their nesting season and how temperature may influence the physiological condition on the offspring. Here we employed biochemical indices based on nucleic acid derived indices (standardized RNA/DNA ratio-sRD, RNA concentration and DNA concentration) in skin tissue as a potential predictor of recent growth rate in nesting females and hatchling loggerhead turtles. Our major findings were that the physiological condition of all nesting females (sRD) decreased during the nesting season, but that females associated with neritic habitats had a higher physiological condition than females associated with oceanic habitats. In addition, the amount of time required for a hatchling to right itself was negatively correlated with its physiological condition (sRD) and shaded nests produced hatchlings with lower sRD. Overall, our results showed that nucleic acid concentrations and ratios of RNA to DNA are an important tool as potential biomarkers of recent growth in marine turtles. Hence, as biochemical indices of instantaneous growth are likely temperature-, size- and age-dependent, the utility and validation of these indices on marine turtles stocks deserves further study.     

Distribution and abundance of marine turtles in the Socialist Republic of Viet Nam

To establish baseline data on the distribution, abundance and threats to marine turtles in Viet Nam we conducted surveys with local fishers, community members and provincial Ministry of Fisheries staff from 17 of Viet Nam’s 29 coastal provinces. These data indicate that five species of marine turtle reside in Viet Nam’s waters (loggerhead, olive ridley, leatherback, green and hawksbill turtles), and four species nest on Viet Nam’s beaches (all of the above except the loggerhead turtle). It is evident from these data that significant declines have occurred in both foraging and nesting populations of all five marine turtle species found in Viet Nam. The greatest current threats to marine turtle populations in Viet Nam are habitat degradation, the accidental and opportunistic of turtles capture by fishers and the direct take of nesting females and their eggs. Successful conservation efforts have been made in recent years through collaboration between international Non Government Organisations and several Vietnamese Government Ministries. Continued success of these projects and the development and implementation of marine conservation policy will depend upon building awareness among Government employees, fishers and the general public about marine turtle biology, ecology, and the need to protect them.     

GLOBAL PHYLOGEOGRAPHY OF THE LOGGERHEAD TURTLE (CARETTA CARETTA) AS INDICATED BY MITOCHONDRIAL DNA HAPLOTYPES

Restriction-site analyses of mitochondrial DNA (mtDNA) from the loggerhead sea turtle (Caretta caretta) reveal substantial phylogeographic structure among major nesting populations in the Atlantic, Indian, and Pacific oceans and the Mediterranean sea. Based on 176 samples from eight nesting populations, most breeding colonies were distinguished from other assayed nesting locations by diagnostic and often fixed restriction-site differences, indicating a strong propensity for natal homing by nesting females. Phylogenetic analyses revealed two distinctive matrilines in the loggerhead turtle that differ by a mean estimated sequence divergence p = 0.009, a value similar in magnitude to the deepest intraspecific mtDNA node (p = 0.007) reported in a global survey of the green sea turtle Chelonia mydas. In contrast to the green turtle, where a fundamental phylogenetic split distinguished turtles in the Atlantic Ocean and the Mediterranean Sea from those in the Indian and Pacific oceans, genotypes representing the two primary loggerhead mtDNA lineages were observed in both Atlantic–Mediterranean and Indian-Pacific samples. We attribute this aspect of phylogeographic structure in Caretta caretta to recent interoceanic gene flow, probably mediated by the ability of this temperate-adapted species to utilize habitats around southern Africa. These results demonstrate how differences in the ecology and geographic ranges of marine turtle species can influence their comparative global population structures.     

Variation in reproductive output of marine turtles

Most marine turtle species are non-annual breeders and show variation in both the number of eggs laid per clutch and the number of clutches laid in a season. Large levels of inter-annual variation in the number of nesting females have been well documented in green turtle nesting populations and may be linked to environmental conditions. Other species of marine turtle exhibit less variation in nesting numbers. This inter-specific difference is thought to be linked to trophic status. To examine whether individual reproductive output is more variable in the herbivorous green turtle (Chelonia mydas Linneaeus 1758) than the carnivorous loggerhead (Caretta caretta Linneaeus 1758), we examined the nesting of both species in Cyprus over nine seasons. Green turtles showed slower annual growth rates (0.11 cm year⁻¹ curved carapace length (CCL) and 0.27 cm year⁻¹ curved carapace width (CCW)) than loggerhead turtles (0.36 cm year⁻¹ CCL, 0.51 cm year⁻¹ CCW). CCL was highly correlated to mean clutch size in both green (R²=0.51) and loggerhead turtles (R²=0.61) and maximal clutch size of green turtles (R²=0.58). Larger females did not lay a greater number of clutches or have a shorter remigration interval than smaller females of either species. On average, the size of green turtle clutches increased and that of loggerhead turtles decreased as the season progressed. Individual green turtles, however, produced more eggs per clutch through the season to a maximum in the third or fourth clutch. In loggerhead turtles, clutches 1-4 were very similar in size but the fifth clutch was 38% smaller than the first. No individuals of either species were recorded laying more than five clutches. Green turtles may not be able to achieve their maximum reproductive output with respect to clutch size throughout the season, whereas only loggerhead turtles laying five clutches (n=5) appear to become resource depleted. Green turtles nesting in years when large numbers of nests were recorded laid a greater number of clutches than females nesting in years with lower levels of nesting.     

Multiple paternity in loggerhead turtle (Caretta caretta) nests on Melbourne Beach, Florida: a microsatellite analysis

Many aspects of sea turtle biology are difficult to measure in these enigmatic migratory species, and this lack of knowledge continues to hamper conservation efforts. The first study of paternity in a sea turtle species used allozyme analysis to suggest multiple paternity in loggerhead turtle (Caretta caretta) clutches in Australia. Subsequent studies indicated that the frequency of multiple paternity varies from species to species and perhaps location to location. This study examined fine-scale population structure and paternal contribution to loggerhead clutches on Melbourne Beach, FL, USA using microsatellite markers. Mothers and offspring from 70 nests collected at two locations were analysed using two to four polymorphic microsatellite loci. Fine-scale population differentiation was not evident between the sampled locations, separated by 8 km. Multiple paternity was common in loggerhead nests on Melbourne Beach; 22 of 70 clutches had more than one father, and six had more than two fathers. This is the first time that more than two fathers have been detected for offspring in individual sea turtle nests. Paternal genotypes could not be assigned with confidence in clutches with more than two fathers, leaving the question of male philopatry unanswered. Given the high incidence of multiple paternity, we conclude that males are not a limiting resource for this central Florida nesting aggregate.     

Monitoring and conservation of critically reduced marine turtle nesting populations: lessons from the Cayman Islands

Historically, nesting marine turtles were abundant in the Cayman Islands and were an integral part of the economy and culture. Today, nesting of loggerhead turtle Caretta caretta and green turtles Chelonia mydas takes place at very low levels. Hawksbill Eretmochelys imbricata nesting has not been recorded since 1999. We overview highly detailed monitoring data gathered over a 6-year period allowing insight into the magnitude and spatial and temporal patterns of marine turtle nesting, cost-effectiveness of monitoring such reduced populations, impacts of development on reproductive success and current threats to the recovery of the population. Nesting is diffuse and widely distributed for both nesting species on Grand and Little Cayman. Modelled nesting detection profiles for Grand Cayman show that in order to maintain data resolution, most sandy coastline must be surveyed throughout each season. However, in Little Cayman it may be possible to reduce effort. Legal take of adults and illegal take of eggs may be significantly impacting the remaining population. Surprisingly, we observed no significant correlation between density of coastal development and clutch density, adult emergence success or hatching success for either species. A significant relationship exists however, between density of coastal development and incidence of misorientation events in loggerhead hatchlings but not in green turtle hatchlings. Effective protection of known nesting habitat and the elimination of exploitation of remaining adults and eggs within the population are critical to its recovery.     

Multiple paternity in egg clutches of hawksbill turtles (Eretmochelys imbricata)

We present the first data collected on the genetic mating system of the hawksbill turtle Eretmochelys imbricata, the only marine turtle not studied to date. We examined paternity within 12 egg clutches from ten female hawksbill turtles from Sabah Turtle Islands, Malaysia. A total of 375 hatchlings were analysed using five microsatellite markers. Results demonstrated that clutches from two out of ten females were sired by multiple males (maximum of two). Although at a low frequency, observation of multiple paternity indicates that hawksbill turtles exhibit the same genetic mating system (polyandry) as observed for other species of marine turtles. Consistent paternity across multiple clutches laid by individual females in one breeding season supports the hypothesis that sperm are stored from mating prior to nesting and are then used to fertilize all subsequent clutches of eggs that season.     

Molecular resolution of marine turtle stock composition in fishery bycatch: a case study in the Mediterranean

Based on an extensive sampling regime from both nesting populations and bycatch, frequency analyses of mitochondrial (mt) DNA control region haplotypes in the Mediterranean were used to assess the genetic structure and stock composition of the loggerhead sea turtle, Caretta caretta, in different marine fisheries. The analyses show the following. (i) In drifting longline fisheries working in Mediterranean pelagic habitats 53–55% of turtles caught originated from the Mediterranean stock; (ii) In bottom-trawl fisheries all turtle bycatch is derived from this regional stock; (iii) This regional stock contribution to fishery bycatch suggests that the population size of the Mediterranean loggerhead nesting population is significantly larger than previously thought. This is consistent with a recent holistic estimate based on the discovery of a large rookery in Libya. (iv) Present impact of fishery-related mortality on the Mediterranean nesting population is probably incompatible with its long-term conservation. Sea turtle conservation regulations are urgently needed for the Mediterranean fisheries. (v) The significant divergence of mtDNA haplotype frequencies of the Turkish loggerhead colonies define this nesting population as a particularly important management unit. Large immature and adult stages from this management unit seem to be harvested predominantly by Egyptian fisheries. (vi) Combined with other data, our findings suggest that all the nesting populations in the Mediterranean should be considered as management units sharing immature pelagic habitats throughout the Mediterranean (and possibly the eastern Atlantic), with distinct and more localized benthic feeding habitats in the eastern basin used by large immatures and adults. (vii) Between the strict oceanic pelagic and the benthic stages, immature turtles appear to live through an intermediate neritic stage, in which they switch between pelagic and benthic foods.     

Avian Incubation Inhibits Growth and Diversification of Bacterial Assemblages on Eggs

Microbial infection is a critical source of mortality for early life stages of oviparous vertebrates, but parental defenses against infection are less well known. Avian incubation has been hypothesized to reduce the risk of trans-shell infection by limiting microbial growth of pathogenic bacteria on eggshells, while enhancing growth of commensal or beneficial bacteria that inhibit or competitively exclude pathogens. We tested this hypothesis by comparing bacterial assemblages on naturally incubated and experimentally unincubated eggs at laying and late incubation using a universal 16S rRNA microarray containing probes for over 8000 bacterial taxa. Before treatment, bacterial assemblages on individual eggs from both treatment groups were dissimilar to one another, as measured by clustering in non-metric dimensional scaling (NMDS) ordination space. After treatment, assemblages of unincubated eggs were similar to one another, but those of incubated eggs were not. Furthermore, assemblages of unincubated eggs were characterized by high abundance of six indicator species while incubated eggs had no indicator species. Bacterial taxon richness remained static on incubated eggs, but increased significantly on unincubated eggs, especially in several families of Gram-negative bacteria. The relative abundance of individual bacterial taxa did not change on incubated eggs, but that of 82 bacterial taxa, including some known to infect the interior of eggs, increased on unincubated eggs. Thus, incubation inhibits all of the relatively few bacteria that grow on eggshells, and does not appear to promote growth of any bacteria.     

Impacts of a geotextile container dune core on marine turtle nesting in Juno Beach,Florida, United States

Hard armoring technologies (e.g. rock revetments and seawalls), which are installed to protect homes from beach erosion, can diminish the aesthetics and amenity of the beach. Over time, these structures cause beach narrowing and often prevent marine turtle access to nesting habitat altogether. An alternative armoring technology, known as geotextile dune core systems (or geocores), has been developed and implemented to protect inland infrastructure from beach erosion, yet there remains an absence of research on possible effects on marine turtles. In this study, we examined the impacts of a geocore installed on Juno Beach, Florida, United States in February 2014 on loggerhead (Caretta caretta) and green turtle (Chelonia mydas) nesting success, hatching success, and emergence success. A before‐after‐control‐impact paired series design evaluated the difference in nesting success per week for the impact and control sites 4 years before (2010–2013) and 4 years after (2014–2017) the installation of the geocore. Neither loggerhead nor green turtle nesting success was significantly different after the installation of the geocore; however, when analyzing loggerhead crawls that came to within 5 m of the geotextile bags, nesting success decreased. Neither hatching nor emergence success was significantly different after the installation of the geocore for either species. Our results suggest that geocores may minimally affect loggerhead and green turtles and provide a suitable restoration technique for homeowners facing beach erosion.     

Accounting for Imperfect Detection Is Critical for Inferring Marine Turtle Nesting Population Trends

Assessments of population trends based on time-series counts of individuals are complicated by imperfect detection, which can lead to serious misinterpretations of data. Population trends of threatened marine turtles worldwide are usually based on counts of nests or nesting females. We analyze 39 years of nest-count, female-count, and capture-mark-recapture (CMR) data for nesting loggerhead turtles (Caretta caretta) on Wassaw Island, Georgia, USA. Annual counts of nests and females, not corrected for imperfect detection, yield significant, positive trends in abundance. However, multistate open robust design modeling of CMR data that accounts for changes in imperfect detection reveals that the annual abundance of nesting females has remained essentially constant over the 39-year period. The dichotomy could result from improvements in surveys or increased within-season nest-site fidelity in females, either of which would increase detection probability. For the first time in a marine turtle population, we compare results of population trend analyses that do and do not account for imperfect detection and demonstrate the potential for erroneous conclusions. Past assessments of marine turtle population trends based exclusively on count data should be interpreted with caution and re-evaluated when possible. These concerns apply equally to population assessments of all species with imperfect detection.     

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).     

Eggs yield nuclear DNA from egg-laying female cowbirds, their embryos and offspring

Here we report methods forextracting maternal DNA from avian eggshells oroffspring DNA from eggshells and embryos. Thesemethods offer alternative techniques forobtaining DNA from oviparous organisms. UsingDNA extracted from eggshells, we obtainmicrosatellite genotypes of the brood parasiticbrown-headed cowbird (Molothrus ater)female that laid the eggs and/or her hatchedoffspring. Using DNA extracted from embryos, weobtain microsatellite genotypes of offspring.We demonstrate that separate extractionsperformed on the embryo and shell from a singleegg can provide DNA from the embryo and itsmother, respectively. This single-egg approachfor obtaining both maternal and embryonic DNAsimplifies paternity analyses because allelesunique to the embryo can be considered paternalin origin. Finally we report two newmicrosatellite loci and primer sequences forbrown-headed cowbirds.     

Natural beaches confer fitness benefits to nesting marine turtles

Coastal ecosystems provide vital linkages between aquatic and terrestrial habitats and thus support extremely high levels of biodiversity. However, coastlines also contain the highest densities of human development anywhere on the planet and are favoured destinations for tourists, creating a situation where the potential for negative effects on coastal species is extremely high. I gathered data on marine turtle reproductive output from the literature to determine whether coastal development negatively influences offspring production. Female loggerhead (Caretta caretta) and green turtles (Chelonia mydas) nesting on natural beaches (as opposed to beaches with permanent development) produce significantly more hatchling turtles per nest; all else being equal, females that successfully produce more offspring will have higher fitness than conspecifics producing fewer offspring. Thus, female marine turtles nesting on natural beaches probably have higher fitness than turtles nesting on developed beaches. Consequently, populations nesting on natural beaches may be able to recover more quickly from the historic population declines that have plagued marine turtles, and some species may recover more quickly than others.     

Incubation reduces microbial growth on eggshells and the opportunity for trans-shell infection

Avian eggshells harbour microbes shortly after laying, and under appropriate ambient conditions they can multiply rapidly, penetrate through shell pores, infect egg contents and cause embryo mortality. We experimentally examined how incubation affects bacterial processes on the eggshells of pearl-eyed thrashers Margarops fuscatus nesting in tropical montane and lowland forests in Puerto Rico. Bacteria and fungi grew rapidly on shells of newly laid, unincubated eggs exposed to ambient conditions, but declined to low levels on shells of eggs incubated by thrashers. Divergence in bacterial growth between incubated and exposed eggs was more marked at the montane forest than at the lowland site. Pathogenic microorganisms became increasingly dominant on shells of exposed eggs, but these groups were relatively rare on incubated eggs, where more benign, less invasive groups prevailed. Some incubation during laying may be necessary to decrease the probability of trans-shell infection by reducing the growth of harmful bacteria and fungi on eggshells, although it may increase hatching asynchrony and the likelihood of brood reduction.     

Twelve new polymorphic microsatellite markers from the loggerhead sea turtle (Caretta caretta) and cross-species amplification on other marine turtle species

We describe 12 new polymorphic dinucleotide microsatellite loci and multiplex Polymerase Chain Reaction conditions from the loggerhead sea turtle Caretta caretta. Levels of polymorphism were assessed in 50 individuals from the nesting population of the Cape Verde Islands. Number of alleles ranged from 3 to 13 (average of 7.33) and the values of observed heterozygosities from 0.32 to 0.80 (average of 0.61). Cross-species amplification on three other marine turtles, Chelonia mydas, Eretmochelys imbricata and Dermochelys coriacea, revealed polymorphism and variability at eight, eleven and three loci, respectively.     

Some Aspects of the Nesting Behavior and Reproductive Biology of Sea Turtles

Basic reproductive data from 21 green turtle (Chelonia mydas),8 leatherback (Dermochelys coriacea), 7 hawksbill (Eretmochelysimbricata), 7 olive ridley (Lepidochelys olivacea),6 loggerhead(Caretta caretta), 1 Kemp's ridley (Lepidochelys kempi), and1 flatback (Chelonia depressa) populations are provided. Someintraspecific and interspecific relationships between size ofnester and clutch, egg size and hatchling size are analyzed.Measurements of reproductive rates (=numbers of hatchlings perfemale per year) in 11 populations varied from 35 to 200 inan olive ridley and loggerhead colony, respectively. Nestingbehavior of each species is described in terms of type of nestingemergence and time spent on the nesting beach (=chelonery).The relatively large number of yolkless eggs laid by many leatherbacksand by some hawksbills invites further study. Some aspects ofsea turtle nesting behavior and reproduction are compared tothose of other chelonians.     

Stormy oceans are associated with declines in sea turtle hatching

Many sea turtle populations are below 10% of their pre-Columbian numbers [1-4]. Though historic and systematic over-exploitation is the principal cause of these declines, sea turtles face similar threats today. Adults and juveniles are actively hunted and commercial fisheries catch them incidentally. Nesting suffers from beach development, egg poaching and the poaching of nesting females. Accompanying these familiar hazards is the largely unknown consequences of recent climate change. Here we report monitoring surveys from the Dry Tortugas National Park (DTNP, 24.64N 82.86W), Florida, and show that hurricanes and other storm events are an additional and increasing threat to loggerhead turtle (Caretta caretta) and green sea turtle (Chelonia mydas) nesting. Both species are listed by the US Endangered Species Act and the IUCN considers them 'endangered'.     

Genetic structure of loggerhead turtle (Caretta caretta) populations in Turkey

This study investigates genetic structure of loggerhead turtle (Caretta caretta) for conservation purposes using both mitochondrial and nuclear DNA markers. To evaluate overall genetic structure of C. caretta at the Turkish nesting beaches and to compare it with previously published data from other nesting sites of the Mediterranean Sea, we have studied 256 hatchlings from 18 different nesting beaches. Seven distinct haplotypes were detected, of which three have previously been reported from the Mediterranean and one from the Atlantic. The remaining three haplotypes are described for the first time for C. caretta in the present study. Distribution of these haplotypes among the nesting sites showed a significant genetic structuring, indicating that females are philopatric and that gene flow among populations is restricted. Both mtDNA and microsatellite analyses determined genetic structuring (mtDNA: γ_st = 0.214, p < 0.01; nDNA F_st = 0.0004 p < 0.05) among nesting aggregates of C. caretta throughout the study area and enabled the detection of the different haplotypes to inform conservation strategies.     

Communal egg-laying behaviour and the consequences of egg aggregation in the brown anole (Anolis sagrei)

Communal nesting is a behaviour that involves multiple females laying eggs in the same nest or nesting site. This behaviour may be a consequence of a shortage of preferable nest sites (constraint hypothesis) or an adaptation generated by fitness benefits associated with egg aggregation (adaptive hypothesis). Experimental tests of these hypotheses require information about maternal nest site choice and its fitness consequences. To address these, we studied a lizard (brown anole; Anolis sagrei) that produces single-egg clutches, but often aggregates eggs in nest sites. In a lab study, females were given the option of nesting in (a) soil previously used as nest substrate vs. fresh soil and (b) soil with eggshells vs. without eggshells. We also experimentally examined the effects of egg aggregation by incubating eggs singly, in groups of four, and in groups of nine. We recorded egg surface temperature, water uptake, and hatchling morphology. Females were more likely to nest in pots with used soil and with eggshells than in pots with fresh soil or without eggshells. We observed no effects of egg aggregation on egg survival, egg temperature, or most measures of hatchling morphology. However, singly-incubated eggs absorbed more water than eggs incubated in the four and nine egg aggregations and this resulted in offspring with greater body condition (i.e. heavier for their length) at hatching. The behavioural experiment demonstrates that females actively choose nest sites that have been used previously (as expected under the adaptive hypothesis), but the egg-aggregation experiment shows no benefits to offspring based on the variables measured. Thus, results of the behaviour study support the adaptive hypothesis; however, results from our egg-incubation study do not. Likely, the adaptive and constraint hypotheses are not mutually exclusive, and a diversity of factors influence the evolution of communal nesting behaviour.