Combining stable isotopes and skeletal growth marks to detect habitat shifts in juvenile loggerhead sea turtles Caretta caretta

Understanding the phase and timing of ontogenetic habitat shifts underlies the study of a species’ life history and population dynamics. This information is especially critical to the conservation and management of threatened and endangered species, such as the loggerhead sea turtle Caretta caretta. The early life of loggerheads consists of a terrestrial egg and hatchling stage, a posthatchling and juvenile oceanic, pelagic feeding stage, and a juvenile neritic, primarily benthic feeding stage. In the present study, novel approaches were applied to explore the timing of the loggerhead ontogenetic shift from pelagic to benthic habitats. The most recent years of somatic growth are recorded as annual marks in humerus cross sections. A consistent growth mark pattern in benthic juvenile loggerheads was identified, with narrow growth marks in the interior of the bone transitioning to wider growth marks at the exterior, indicative of a sharp increase in growth rates at the transitional growth mark. This increase in annual growth is hypothesized to correlate with the ontogenetic shift from pelagic to benthic habitats. Stable isotopes of carbon and nitrogen just interior and exterior to the transitional growth mark, as well as stable isotopes from pelagic and benthic flora, fauna and loggerhead stomach contents, were analyzed to determine whether this transition related to a diet shift. The results clearly indicate that a dietary shift from oceanic/pelagic to neritic/benthic feeding corresponds to a transitional growth mark. The combination of stable isotope analysis with skeletochronology can elucidate the ecology of cryptic life history stages during loggerhead ontogeny.

Use of skeletochronological analysis to estimate the age of leatherback sea turtles Dermochelys coriacea in the western North Atlantic

Although growth rate and age data are essential for leatherback management, estimates of these demographic parameters remain speculative due to the cryptic life history of this endangered species. Skeletochronological analysis of scleral ossicles obtained from 8 captive, known-age and 33 wild leatherbacks originating from the western North Atlantic was conducted to characterize the ossicles and the growth marks within them. Ages were accurately estimated for the known-age turtles, and their growth mark attributes were used to calibrate growth mark counts for the ossicles from wild specimens. Due to growth mark compaction and resorption, the number of marks visible at ossicle section tips was consistently and significantly greater than the number visible along the lateral edges, demonstrating that growth mark counts should be performed at the tips so that age is not underestimated. A correction factor protocol that incorporated the trajectory of early growth increments was used to estimate the number of missing marks in those ossicles exhibiting resorption, which was then added to the number of observed marks to obtain an age estimate for each turtle. A generalized smoothing spline model, von Bertalanffy growth curve, and size-at-age function were used to obtain estimates of age at maturity for leatherbacks in the western North Atlantic. Results of these analyses suggest that median age at maturation for leatherbacks in this part of the world may range from 24.5 to 29 yr. These age estimates are much greater than those proposed in previous studies and have significant implications for population management and recovery.

Incidental capture of loggerhead (Caretta caretta) and leatherback (Dermochelys coriacea) sea turtles by the pelagic longline fishery off southern Brazil

Incidental capture in fishing gear is one of the main sources of injury and mortality of juvenile and adult sea turtles (NRC, 1990; Lutcavage et al., 1997; Oravetz, 1999). Six out of the seven extant species of sea turtles — the leatherback (Dermochelys coriacea), the green turtle (Chelonia mydas), the loggerhead (Caretta caretta), the hawksbill (Eretmochelys imbricata), the olive ridley (Lepidochelys olivacea), and the Kemp’s ridley (Lepidochelys kempii) — are currently classified as endangered or critically endangered by the World Conservation Union (IUCN, formerly the International Union for Conservation of Nature and Natural Resources), which makes the assessment and reduction of incidental capture and mortality of these species in fisheries priority conservation issues (IUCN/Species Survival Commission, 1995).

Validation and interpretation of annual skeletal marks in loggerhead (Caretta caretta) and Kemp’s ridley (Lepidochelys kempii) sea turtles

Numerous studies have applied skeletochronology to sea turtle species. Because many of the studies have lacked validation, the application of this technique to sea turtle age estimation has been called into question. To address this concern, we obtained humeri from 13 known-age Kemp’s ridley (Lepidochelys kempii) and two loggerhead (Caretta caretta) sea turtles for the purposes of examining the growth marks and comparing growth mark counts to actual age. We found evidence for annual deposition of growth marks in both these species. Corroborative results were found in Kemp’s ridley sea turtles from a comparison of death date and amount of bone growth following the completion of the last growth mark (n=76). Formation of the lines of arrested growth in Kemp’s ridley sea turtles consistently occurred in the spring for animals that strand dead along the mid- and south U.S. Atlantic coast. For both Kemp’s ridley and loggerhead sea turtles, we also found a proportional allometry between bone growth (humerus dimensions) and somatic growth (straight carapace length), indicating that size-at-age and growth rates can be estimated from dimensions of early growth marks. These results validate skeletochronology as a method for estimating age in Kemp’s ridley and loggerhead sea turtles from the southeast United States.

Estimates of survival probabilities for oceanic-stage loggerhead sea turtles (Caretta caretta) in the North Atlantic

Estimates of instantaneous mortality rates (Z) and annual apparent survival probabilities (Φ) were generated from catch-curve analyses for oceanic-stage juvenile loggerheads (Caretta caretta) in the waters of the Azores. Two age distributions were analyzed: the “total sample” of 1600 loggerheads primarily captured by sighting and dipnetting from a variety of vessels in the Azores between 1984 and 1995 and the “tuna sample” of 733 loggerheads (a subset of the total sample) captured by sighting and dipnetting from vessels in the commercial tuna fleet in the Azores between 1990 and 1992. Because loggerhead sea turtles begin to emigrate from oceanic to neritic habitats at age 7, the best estimates of instantaneous mortality rate (0.094) and annual survival probability (0.911) not confounded with permanent emigration were generated for age classes 2 through 6. These estimates must be interpreted with caution because of the assumptions upon which catch-curve analyses are based. However, these are the first directly derived estimates of mortality and survival probabilities for oceanic-stage sea turtles. Estimation of survival probabilities was identified as “an immediate and critical requirement” in 2000 by the Turtle Expert Working Group of the U.S. National Marine Fisheries Service.

The physiological effects of multiple forced submergences in loggerhead sea turtles (Caretta caretta)

Sea turtles are subjected to involuntary submergence and potential mortality due to incidental capture by the commercial shrimp fishing industry. Despite implementation of turtle excluder devices (TEDs) to reduce at-sea mortality, dead stranded turtles continue to be found in near-record numbers along the coasts of the western Atlantic Ocean and northern Gulf of Mexico. Although this mortality may be due to an increase in the number of turtles available to strand, one alternative explanation is that sea turtles are repetitively submerged (as one fishing vessel follows the path of another) in legal TEDs. In the present study, laboratory and field investigations were undertaken to examine the physiological effects of multiple submergence of loggerhead sea turtles (Caretta caretta). Turtles in the laboratory study were confined during the submersion episodes, whereas under field conditions, turtles were released directly into TED-equipped commercial fishing nets. Under laboratory and field conditions, pre- and postsubmergence blood samples were collected from turtles submerged three times at 7.5 min per episode with an in-water rest interval of 10, 42, or 180 min between submergences. Analyses of pre- and postsubmergence blood samples revealed that the initial submergence produced a severe and pronounced metabolic and respiratory acidosis in all turtles. Successive submergences produced significant changes in blood pH, Pco2, and lactate, although the magnitude of the acid-base imbalance was substantially reduced as the number of submergences increased. In addition, increasing the interval between successive submergences permitted greater recovery of blood homeostasis. No turtles died during these studies. Taken together, these data suggest that repetitive sub-mergence of sea turtles in TEDs would not significantly affect their survival potential provided that the animal has an adequate rest interval at the surface between successive submergences.

Origin of immature loggerhead sea turtles (Caretta caretta) at Hutchinson Island, Florida: evidence from mtDNA markers

Loggerhead sea turtles (Caretta caretta) are migratory, long-lived, and slow maturing. They are difficult to study because they are seen rarely and their habitats range over vast stretches of the ocean. Movements of immature turtles between pelagic and coastal developmental habitats are particularly difficult to investigate because of inadequate tagging technologies and the difficulty in capturing significant numbers of turtles at sea. However, genetic markers found in mitochondrial DNA (mtDNA) provide a basis for predicting the origin of juvenile turtles in developmental habitats. Mixed stock analysis was used to determine which nesting populations were contributing individuals to a foraging aggregation of immature loggerhead turtles (mean 63.3 cm straight carapace length [SCL]) captured in coastal waters off Hutchinson Island, Florida. The results indicated that at least three different western Atlantic loggerhead sea turtle subpopulations contribute to this group: south Florida (69%), Mexico (20%), and northeast Florida-North Carolina (10%). The conservation and management of these immature sea turtles is complicated by their multinational genetic demographics.

Water temperature and internesting intervals for loggerhead (Caretta caretta) and green (Chelonia mydas) sea turtles

Temperature loggers were attached to the carapace of green turtles (Chelonia mydas) at Ascension Island and Cyprus and to loggerhead turtles (Caretta caretta) at Cyprus, in order to record the ambient temperature experienced by individuals during the internesting interval, i.e. the period between consecutive clutches being laid. Internesting intervals were relatively short (10-14 days) and mean ambient temperatures relatively warm (27-28degreesC), compared to previous observations for these species nesting in Japan, although a single internesting interval versus temperature relationship described all the data for these two species from the different areas. The implication is that water temperature has both a common and a profound effect on the length of the internesting interval for these two species: internesting intervals are shorter when the water is warmer. (C) 2002 Elsevier Science Ltd. All rights reserved.

Gonad development and hormone titres in Loggerhead Sea Turtles (Caretta caretta) in the NE Atlantic

The study proposed to describe sexual development in pelagic stage loggerhead sea turtles Caretta caretta and compare this to hatchlings and adults. It is meant as an ontogenic approach, in order to understand reproductive development and population composition and their dynamics in the pelagic environment. The study focused on the pelagic loggerheads that are found in the waters offshore Madeira Island (Portugal) in the North-eastern Atlantic and use it as a developmental habitat. The innovating character of this work relied on the lack of any description regarding the gonad ontogenesis and reproductive development for the pelagic stage in any of the 7 existing sea turtle species, all of them in danger of extinction. Three methods were used to diagnose the sex of each juvenile individual and asses the level of reproductive development: (1) laparoscopy, (2) gonad biopsy and (3) the assessment of two sex steroids circulating levels, namely testosterone and estradiol. In order to cover all life stages and compare data obtained for the juvenile stage, hatchlings and nesting female adults were sampled at the nearest nesting rookery at Boa Vista Island in the Cape Verde Archipelago. Gonads from dead hatchlings were collected for gonad histology and blood was collected from nesting females for sex steroids assessment. Laparoscopies revealed to be a valid sexing method for the juvenile stage, since gonads are morphologically differentiated at these size classes. Moreover, laparoscopy was validated using gonad histology. Gonad histology of juveniles showed that gonads are already completely differentiated into ovaries or testes at the size classes examined, but development seems to be quiescent. Males present already developed seminiferous tubules with spermatogonia lining the interior of the seminiferous tubule. Female gonads present oocytes at different development stages, but only oocytes up to stage III were observed. The maximum oocyte diameter in each individual correlated with body size, suggesting that reproductive development is an on-going process in juvenile females. The circulating levels of both testosterone and estradiol in juveniles of both sexes were very low and consistently lower than the ones observed in the nesting females from Boa Vista Island. No bimodal distribution was found for any of the sex steroids analysed and thus circulating hormone levels were not a reliable tool for sexing juvenile individuals with a non-invasive technique. The ratio testosterone:estradiol did not show a bimodal distribution either. The levels of testosterone correlated with sea surface temperature. The fact that temperatures observed during this study were below 24ºC might have hindered a differential testosterone pattern between juvenile males and females. Sex ratios for this population were generated according to laparoscopy results and compared among years and size classes. An overall sex ratio of 2 females for each male was found, but they varied among size classes but not among years. Possible causes for the sex ratios observed are discussed. This study is a contribution to our knowledge on the pelagic stage of loggerhead turtles, namely on the population structure regarding sex ratio, which is a vital tool for implementing conservation strategies.

Contribution to the chemoreception capacity of juvenile Loggerhead sea turtles (Caretta caretta, L.)

Loggerhead sea turtle juveniles (Caretta caretta), pelagic stage, are found in waters of Madeira archipelago. Pelagic turtles are in the main growth phase of their life cycle and consequently higher energy needs. However, knowledge about the ecology of pelagic loggerhead sea turtles is still quite rudimentary, mainly about the mechanisms that lead them to find food in the vast ocean. Studies with other pelagic species, such as procellariiform birds, revealed that the olfactory system play an important role for the detection of feeding areas, through the detection of concentration peaks of DMS (dimethylsulfide), a scent compound that naturally exists in the marine environment and it is related to areas of high productivity. Based on the assumption that loggerhead sea turtles use a similar mechanism, behavioural experiments were conducted in order to analyze the chemoreception capacity to DMS (airborne chemoreception - theoretically responsible for the long distance detection of areas with food patches; and aquatic chemoreception - theoretically responsible for the short distance detection of preys). The first step was to observe if pelagic loggerheads demonstrate sensitivity to DMS and the second was to verify if they really use the DMS, in natural conditions, as an airborne cue to find areas where food patches might be available. Four juveniles of loggerhead sea turtles were tested in captivity and three wild turtles in the open ocean. The results of airborne chemoreception experiments in captivity revealed that one turtle clearly demonstrated sensitivity to DMS and the sea experiments confirmed this result. However, the experiments were not conclusive on the question whether the pelagic turtles actually use the DMS as an airborne cue to detect long distance food patches. In aquatic chemoreception experiments was not observed sensitivity to DMS by the three sea turtles tested. In the classical conditioning experiment, where DMS and food were given nearly at the same time revealed that after a certain period of time, the sea turtle tested did not associated the DMS stimulus with a possible food reward. The main cause of mortality of loggerhead sea turtles in Madeira waters is due to the accidental capture (bycatch) by deep pelagic longlines fishery which the target species is the black-scabbard (Aphanopus carbo) fish. Chub mackerel (Scomber japonicus) is one of the baits used in this fishery. Aquatic chemoreception experiments were conducted in order to evaluate the attractiveness of the chub mackerel for sea turtles. For the three sea turtles tested, the results showed that in 90% of the cases the sea turtles were extremely attracted by the underwater smell of this fish.