Metabolic heating in Mediterranean loggerhead sea turtle clutches

Offspring sex ratio is an important demographic parameter and, given its determination by incubation temperature in sea turtles, might be a key factor for their conservation under climate warming. An appealing approach to estimate hatchling sex ratios is to measure sand temperatures at nest depth and deduce hatchling sex ratios from a beforehand-established relationship of hatchling sex ratio and sand temperature. Such estimates will only be accurate though if metabolic heat produced by the embryos is considered. Judging whether metabolic heating has a potential effect on hatchling sex ratios without actually measuring temperature within clutches would greatly facilitate monitoring protocols. We tested for a relationship between the amount of metabolic heating and the number of developed embryos as well as clutch size in the largest known loggerhead sea turtle (Caretta caretta) population of the Mediterranean on Zakynthos (Greece). Temperatures were measured within 20 nests as well as at a reference site in the sand at nest depth. Metabolic heating was detected, but only during the last third of the incubation period did nests heat up considerably (1.6 °C on average) above the temperature of the surrounding sand. During the middle third of incubation, when sex is determined, the amount of metabolic heating was negligible. The amount of metabolic heating during the last third of the incubation duration was significantly correlated to the number of offspring developed to at least about 75% of incubation duration. This factor explained nearly 50% of variation in metabolic heating. Metabolic heating was also significantly correlated to clutch size. Given that clutch size within the Mediterranean is largest in Zakynthos loggerheads, we conclude that metabolic heating can be ignored in the estimate of hatchling sex ratios in Mediterranean loggerhead populations. These results thus provide the basis for a feasible monitoring of hatchling sex ratios in the loggerhead sea turtle in the Mediterranean.     

Post-nesting movements and submergence patterns of loggerhead marine turtles in the Mediterranean assessed by satellite tracking

Two female loggerhead turtles (Caretta caretta) were tracked, following nesting at Alagadi Beach (35°33′N, 33°47′E), Northern Cyprus, eastern Mediterranean for 60 and 82 days, respectively. The two individuals showed marked differences in their behaviour. Individual A was tracked to Syrian coastal waters, whereas individual B travelled around the coast of Northern Cyprus to a foraging site in the waters off the east coast of Northern Cyprus. Submergence durations varied markedly during different phases of the migration, suggesting coastal foraging/resting at certain stages en route with sustained directed travelling movements during initial coastal movements and open ocean crossing. Both turtles showed fidelity to foraging grounds for the duration of transmissions (Turtle A: 36 days; Turtle B: 58 days). In both cases, locations were centred in inshore waters although the two individuals exhibited quite different submergence patterns. Individual A carried out very short dives of typical duration <10 min, whereas Individual B carried out longer dives with typical duration >20 min. Diel differences in submergence duration at the foraging grounds suggested longer dives at night/early morning for both turtles. For Turtle A, there was a general reduction in submergence duration as the period of residence increased; a pattern that may have been related to increasing temperature experienced. The total distance travelled by the two turtles (320 and 227 km) was relatively short when compared to satellite tracking studies of conspecifics following nesting in South Africa and USA and tagging studies of nesting loggerhead turtles in Greece and Australia. It is hypothesized that short migratory distance may be correlated with both the small body size and the relatively high frequency of remigration in this population.     

Establishing physiological blood parameters in the loggerhead sea turtle (Caretta caretta)

The loggerhead sea turtle (Caretta caretta), one of the three sea turtle species inhabiting the Mediterranean Sea, is an endangered species. However, although frequently treated in marine animal rescue centers, and the subject of several studies in literature, as yet, few studies have been conducted on large numbers of loggerhead sea turtles in order to establish physiological reference ranges that enable the identification of pathological values. The lack of studies on reference parameters probably depends on the fact that marine turtles treated in wildlife rescue centers are usually in a critical conditions, thus precluding the collection of data on healthy animals and compromising the reliability of any data obtained from them. The present biological study was therefore conducted in order to obtain a database from healthy animals in natural conditions. Serum biochemistry and serum protein electrophoresis were performed on blood samples obtained from 65 healthy adult loggerhead sea turtles captured and delivered to the Sea Turtle Rescue Center of Linosa (Italy); the blood samples were collected during the clinical examination of rescued animals. Laboratory analyses of serum samples were made in order to establish reference parameters, commonly required for laboratory diagnoses in mammals and diseased animals.

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).     

Unfolding of the loggerhead sea turtle (Caretta caretta) myoglobin: A (1)H-NMR and electronic absorbance study

The effect of urea concentration on the backbone solution structure of the cyanide derivative of ferric Caretta caretta myoglobin (at pH 5.4) is reported. By addition of urea, sequential and long-range nuclear Overhauser effects (NOEs) are gradually lost. By using the residual NOE constraints to build the molecular model, a picture of the unfolding pathway was obtained. When the urea concentration is raised to 2.2 M, helices A and B appear largely disordered; helices C, D, and F loose structural constraints at 3.0 M urea. At urea concentration >6 M, the protein appears to be fully unfolded, including the GH hairpin and helix E stabilizing the prosthetic group. Reversible and cooperative denaturation isotherms obtained by following NOE peaks are considerably different from those obtained by monitoring electronic absorption changes. The reversible and cooperative urea-dependent folding-unfolding process of C. caretta myoglobin follows the minimum three-state mechanism N long left and right arrow X long left and right arrow D, where X represents a disordered globin structure (occurring at approximately 4 M urea) that still binds the heme.     

The genetic structure of the loggerhead sea turtle (Caretta caretta) in the Mediterranean as revealed by nuclear and mitochondrial DNA and its conservation implications

The population genetic structure of the loggerhead sea turtle (Caretta caretta) nesting in the eastern Mediterranean was assessed by sequencing a fragment of the control region of the mitochondrial DNA (n = 190) and seven microsatellites (n = 112). The two types of markers revealed genetic structuring (mtDNA: γ_st = 0.212, P < 0.001; nDNA F _st = 0.006, P < 0.001), thus indicating that both females and males are philopatric and that gene flow between populations is restricted. Mitochondrial DNA data indicate that the female populations nesting on the islands of Crete and Cyprus have suffered a recent bottleneck or colonization event. However, no bottleneck or founder effect was revealed by nuclear markers, thus indicating male-mediated gene flow from other populations that would increase nuclear genetic variability. Crete, and to a lower extent Cyprus, are thought to play a central role in such male-mediated gene flow that may reduce the negative effect of genetic drift or inbreeding on the small populations of Lebanon and Israel. This population structure indicates that assessing population relevance only on the basis of genetic variability and size would be misleading, as some populations not fulfilling those requirements may play a relevant role in genetic exchange and hence contribute to the overall genetic variability.     

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.     

Discovery of a regular nesting area of loggerhead turtle <Emphasis Type="Italic">Caretta caretta</Emphasis> in southern Italy: a new perspective for national conservation

Loggerhead turtle Caretta caretta nesting in Italy had been reported to be limited to the Pelagian Islands and only sporadically elsewhere. As presence of loggerhead turtle nests had occasionally been reported (1988–1999) along about 200 km of the Ionian coast of Calabria, we carried out a project to assess the actual state of the nesting population between 2000 and 2004. We divided the coastline in two sectors (A: 52 km, and B: 146 km) that were monitored from mid-June to end of July for a total of n = 174 monitoring days and 1,813.6 km patrolled on foot with different intensities (extensive versus intensive). In sector B, through extensive monitoring we did not find any emergence tracks, but in sector A by intensive survey (2002–2004: one survey/3.64 days) we detected 3–8 nests/year. In total, 25 nests (both observed and reported), were recorded in our study area, and an assessment of a total of 15–16 nests/year was suggested. These figures, within the national scenario depicted from the review of known nesting events in the last 40 years (88 records concerning more than 143–144 nests), show that loggerhead turtle nesting has been underestimated in Italy, due to inadequate monitoring protocols, and that nesting is more frequent than expected (at least 30–40 nests/year). Conservation strategies in Italy should then focus not only on the reduction of mortality at sea, but also include large-scale actions to preserve scattered (but regular) nesting events.     

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.     

Using an individual-based model for assessment of sea turtle population viability

Marine turtle species have a complex life history characterized by interannual variability in reproductive performance and a long life span. These ecological features in combination with the animals highly migratory nature create numerous difficulties when trying to assess population dynamics. This study attempts to couple existing information on species demographics and behavioral strategies with simple energetic rules in a theoretical framework. We study sea turtle population dynamics using an individual-based model that incorporates known behavioral-ecological characteristics of the species. Methodology used to design the model was based on the superindividual approach (Scheffer et al. Ecol Model 80:161–170, 1995). We constructed our simulation experiment on a virtual sea turtle population, which was parameterized by using recent literature reviews with emphasis on reproductive parameters of the Mediterranean loggerhead sea turtle population. Switching rules describing critical processes of reproductive performances were established as theoretical functions of efficiency of energy transfer. In order to explore the significance of variable reproductive patterns upon population dynamics and persistence, a series of simulations was performed. The model was also run under fluctuated demographic variables to perform a sensitivity analysis of critical parameters and life-history stages. Based on the specific model parameterization, simulation results show that population persistence was most sensitive to fecundity and to survival at the pelagic juvenile stage. Additionally a surprising finding is the relatively high importance of egg survival in terms of both hatching and hatchling success. We conclude that enhancing the population with new individuals by increasing survival in the early life stages could compensate for additional losses in other age classes. The need for further research regarding biological and behavioral features as well as basic demographic insights into the endangered loggerhead sea turtle is also highlighted.     

Comparative study on the fatty acid composition of two marine vertebrates: striped dolphins and loggerhead turtles

The fatty acid composition of total lipids extracted from seven different tissues (fat, liver, cerebrum, cerebellum, lung, kidney and muscle) of 10 striped dolphins (Stenella coeruleoalba) (weight 75.7±16.2 kg) found dead or moribund during a morbillivirus epizootic, and from two tissues (fat and liver) of 54 loggerhead turtles (Caretta caretta) (18.5±10.5 kg) seized dead after illegal capture, both from the Mediterranean Sea, have been determined by high resolution chromatography techniques. When comparing both species, fatty acid composition is surprisingly similar, with a predominance of the monoenoic followed by the saturated group of fatty acids, and a very close ratio of polyunsaturates n-3/n-6. The relatively high arachidonic acid content in the liver of the two marine species is remarkable. The similar diet can play an important role in these findings, but it is suggested that probably metabolic pathways and essential fatty acid requirements between both marine vertebrates are similar, more than is expected from their earlier filogenetic evolutionary divergence.     

Responses of hatchling sea turtles to rotational displacements

After emerging from underground nests, sea turtle hatchlings migrate through the surf zone and out to the open ocean. During this migration, both waves and water currents can disrupt hatchling orientation by unpredictably rotating the turtles away from their migratory headings. In addition, waves cause turtles to roll and pitch, temporarily impeding forward swimming by forcing the hatchlings into steeply inclined positions. To maintain seaward orientation and remain upright in the water column, hatchlings must continuously compensate for such displacements. As a first step toward determining how this is achieved, we studied the responses of loggerhead (Caretta caretta L.) sea turtle hatchlings to rotational displacements involving yaw, roll, and pitch. Hatchlings responded to rotations in the horizontal plane (yaw) by extending the rear flipper on the side opposite the direction of rotation. Thus, the flipper presumably acts as a rudder to help turn the turtle back toward its original heading. Turtles responded to rotations in the roll plane with stereotypic movements of the front flippers that act to right the hatchlings with respect to gravity. Finally, hatchlings responded to rotations in the pitch plane with movements of the hind flippers that appear likely to curtail or counteract the pitching motion. Thus, the results of these experiments imply that young sea turtles emerge from their nests possessing a suite of stereotypic behavioral responses that function to counteract rotational displacements, enable the animals to maintain equilibrium, and facilitate efficient movement toward the open sea.     

Invertebrate infestation on eggs and hatchlings of the loggerhead turtle, Caretta caretta, in Dalaman, Turkey

The damage caused by some invertebrates to the eggs and hatchlings of loggerhead turtles, Caretta caretta, was investigated during the summer of 2002 on Dalaman beach, Turkey. The specimens, identified to family or genus levels, from nine families representing seven orders were recorded as infesting nests of loggerhead turtles. The heaviest impacts on loggerhead turtle nests was made by Pimelia sp. (Tenebrionidae, Coleoptera). Twenty-four (36.3%) out of 66 intact loggerhead hatched nests were affected by these larvae. Larval damage by Pimelia sp. was recorded in 188 (10.6%) out of 1773 eggs, but only in two (0.28%) hatchlings. The results show that fewer insects were in the nest the further from vegetation and therefore the relocation of nests from the water's edge to further inland close to vegetation may increase the infestation rate of the eggs.     

Evidence for transoceanic migrations by loggerhead sea turtles in the southern Pacific Ocean

Post-hatchling loggerhead turtles (Caretta caretta) in the northern Pacific and northern Atlantic Oceans undertake transoceanic developmental migrations. Similar migratory behaviour is hypothesized in the South Pacific Ocean as post-hatchling loggerhead turtles are observed in Peruvian fisheries, yet no loggerhead rookeries occur along the coast of South America. This hypothesis was supported by analyses of the size-class distribution of 123 post-hatchling turtles in the South Pacific and genetic analysis of mtDNA haplotypes of 103 nesting females in the southwest Pacific, 19 post-hatchlings stranded on the southeastern Australian beaches and 22 post-hatchlings caught by Peruvian longline fisheries. Only two haplotypes (CCP1 93% and CCP5 7%) were observed across all samples, and there were no significant differences in haplotype frequencies between the southwest Pacific rookeries and the post-hatchlings. By contrast, the predominant CCP1 haplotype is rarely observed in North Pacific rookeries and haplotype frequencies were strongly differentiated between the two regions (F_st=0.82; p=<0.00001). These results suggest that post-hatchling loggerhead turtles emerging from the southwest Pacific rookeries are undertaking transoceanic migrations to the southeastern Pacific Ocean, thus emphasizing the need for a broader focus on juvenile mortality throughout the South Pacific to develop effective conservation strategies.     

Nest site fidelity and clutch frequency of loggerhead turtles are better elucidated by satellite telemetry than by nocturnal tagging efforts: Implications for stock estimation

Satellite telemetry and ground-based tagging studies are complementary methods to define the spatial and temporal patterns of nesting behavior by migratory sea turtles. Estimates of site fidelity and clutch frequencies are compared for satellite telemetry versus ground truth patrols over a 6 km stretch at a southwest Florida loggerhead (Caretta caretta) rookery. Site fidelity ranged from 1.9 km to 109.1 km for all nests deposited by a female within a season. The mean site fidelity was 28.1 km for all nests, but declined to 16.9 km if omitting the first nest. Nest frequency ranged from 2 to 8 nests per season, with a modal value of 5 nests. Satellite telemetry documented a mean nest frequency of 5.4 nests per female in comparison to 2.2 nests detected by monitoring patrols. The remigrant females had higher clutch frequency, were larger in size, and had higher site fidelity compared to newly tagged females. Satellite telemetry provided improved measurements of site fidelity and reveals a need for revised fecundity estimates. If measures of clutch frequency are representative of loggerhead assemblages nesting elsewhere within the South Florida grouping, the confidence bounds on Western Atlantic loggerhead stocks are approximately 32% lower than currently accounted for annual nesting individuals.     

Sea turtle nesting distributions and oceanographic constraints on hatchling migration

Patterns of abundance across a species''s reproductive range are influenced by ecological and environmental factors that affect the survival of offspring. For marine animals whose offspring must migrate long distances, natural selection may favour reproduction in areas near ocean currents that facilitate migratory movements. Similarly, selection may act against the use of potential reproductive areas from which offspring have difficulty emigrating. As a first step towards investigating this conceptual framework, we analysed loggerhead sea turtle (Caretta caretta) nest abundance along the southeastern US coast as a function of distance to the Gulf Stream System (GSS), the ocean current to which hatchlings in this region migrate. Results indicate that nest density increases as distance to the GSS decreases. Distance to the GSS can account for at least 90 per cent of spatial variation in regional nest density. Even at smaller spatial scales, where local beach conditions presumably exert strong effects, at least 38 per cent of the variance is explained by distance from the GSS. These findings suggest that proximity to favourable ocean currents strongly influences sea turtle nesting distributions. Similar factors may influence patterns of abundance across the reproductive ranges of diverse marine animals, such as penguins, eels, salmon and seals.     

The dual function of the lung in chelonian sea turtles: buoyancy control and oxygen storage

Chelonian sea turtles use their lung as a buoyancy organ and as the major oxygen store when diving, and hence, buoyancy regulation and oxygen consumption can be expected to interact. The buoyancy of seven juvenile loggerhead turtles, Caretta caretta, was determined by measuring directly their underwater weight (M_uw) while they were resting on a freely suspended weighing platform at a depth of 80 cm. Underwater weight was recorded continuously for 2 days for each turtle, followed by another 2 days measurements during which the turtles carried lead weights attached to their carapace, and finally, a last day of measurement after the weights had been removed. Total duration of resting dives (t_r), buoyancy (F_B), total resulting force acting on the resting turtle (F_res) and body volumes were derived from the M_uw data. Turtles were slightly negatively buoyant when resting (F_res=−0.2943 to −0.981 N kg⁻¹) and M_uw increased significantly throughout each apnoeic period, meaning that the turtles progressively lost buoyancy. Pulmonary gas loss was calculated from the rate at which buoyancy decreased, which was significantly slower during the first half of the dive than during the second half of the dive. Resting oxygen consumption rates (V_O2) were calculated from these data assuming that the pulmonary gas loss represents oxygen consumption from the lung. The V_O2 obtained in this way (1.69-4.86 ml O₂ min⁻¹) corresponded well with other published and V_O2 measured previously on loggerhead turtles in the same facility. Using oxygen from the lung affects buoyancy and may have impacts on the diving behaviour.     

Persistently high levels of intersexuality in male-biased amphipod populations

In a study conducted over 30 years ago, a remarkable population of amphipod crustaceans (Gammarus minus) was discovered in which 100% of the females displayed intersex characteristics. Such high levels of intersexuality are atypical amongst gonochoristic crustaceans and current theories relating to sex allocation suggest intermediate (intersex) forms should be competitively selected against. This study set out to confirm the existence of this extraordinary population and to compare it with other G. minus populations in various spring and cave systems across Pennsylvania, Virginia and West Virginia (USA), with varying environmental characteristics in an attempt to elucidate the prevalence and cause of intersexuality in this species. Results from this study verify the continued existence of these highly abundant, intersex female populations of G. minus, with proportions ranging between 73% and 100%. These populations were also associated with highly male-biased sex ratios, which is equally unusual amongst gonochoristic crustaceans. Intersex female specimens had a reduced fecundity compared to normal females, although this result was not statistically significant. Although no feminising microsporidian parasites were observed through histological examination, the presence of a novel feminising parasitic factor cannot be ruled out. The continued vigour of these populations over three decades suggests that anthropomorphic factors are not causing the high levels of intersexuality observed. Intersexuality was absent or very low in frequency (<1%) in most other springs that we investigated. Therefore, G. minus appears to represent an interesting and novel model system for studying sex determination.