Clinical Pathology Reference Intervals for an In-Water Population of Juvenile Loggerhead Sea Turtles (Caretta caretta) in Core Sound, North Carolina, USA

The loggerhead sea turtle (Caretta caretta) is found throughout the waters of the Atlantic, Pacific, and Indian Oceans. It is a protected species throughout much of its range due to threats such as habitat loss, fisheries interactions, hatchling predation, and marine debris. Loggerheads that occur in the southeastern U.S. are listed as “threatened” on the U.S. Endangered Species List, and receive state and federal protection. As part of an on-going population assessment conducted by the National Marine Fisheries Service, samples were collected from juvenile loggerhead sea turtles in Core Sound, North Carolina, between 2004 and 2007 to gain insight on the baseline health of the threatened Northwest Atlantic Ocean population. The aims of the current study were to establish hematologic and biochemical reference intervals for this population, and to assess variation of the hematologic and plasma biochemical analytes by season, water temperature, and sex and size of the turtles. Reference intervals for the clinical pathology parameters were estimated following Clinical Laboratory Standards Institute guidelines. Season, water temperature, sex, and size of the turtles were found to be significant factors of variation for parameter values. Seasonal variation could be attributed to physiological effects of decreasing photoperiod, cooler water temperature, and migration during the fall months. Packed cell volume, total protein, and albumin increased with increasing size of the turtles. The size-related differences in analytes documented in the present study are consistent with other reports of variation in clinical pathology parameters by size and age in sea turtles. As a component of a health assessment of juvenile loggerhead sea turtles in North Carolina, this study will serve as a baseline aiding in evaluation of trends for this population and as a diagnostic tool for assessing the health and prognosis for loggerhead sea turtles undergoing rehabilitation

Patterns of loggerhead turtle ontogenetic shifts revealed through isotopic analysis of annual skeletal growth increments

Ontogenetic changes in resource use often delimit transitions between life stages. Ecological and individual factors can cause variation in the timing and consistency of these transitions, ultimately affecting community and population dynamics through changes in growth and survival. Therefore, it is important to document and understand behavioral and life history polymorphisms, and the processes that drive intraspecific variation in them. To evaluate juvenile loggerhead sea turtle (Caretta caretta) life history variation and to detect shifts in habitat and diet that occur during an oceanic-to-neritic ontogenetic shift, we sequentially analyzed the stable isotope composition of humerus bone growth increments from turtles that stranded dead on Southeastern U.S. beaches between 1997 and 2013 (n = 84). In one-half of the sampled turtles, growth increment-specific nitrogen stable isotope (δ¹⁵N) data showed significant increases in δ¹⁵N values over each turtle's life. These data were used to provide a new line of evidence that juvenile Northwest Atlantic loggerheads exhibit two major ontogenetic shift patterns: discrete shifts (n = 24), which were completed within one year, and facultative shifts (n = 14), which were completed over multiple years (up to five). The mean difference in pre- and post-ontogenetic shift δ¹⁵N values was 4.3‰. Differences in isotopic baselines between neritic and oceanic habitats of the Northwest Atlantic Ocean make it likely these patterns are driven by a coupled change in both habitat and diet, and that facultative shifters utilize both neritic and oceanic resources within transitional growth years. Mean size and age at transition between habitats (54.2 cm straightline carapace length, SCL; 11.98 yr) was within the range of previous estimates and did not differ between discrete and facultative shifters. Our results further expand our understanding of loggerhead sea turtle life history polymorphisms and demonstrate the value of bone tissue analysis to the study of this variation. Sequential analysis of annual skeletal growth increments provides a valuable method for reconstructing long-term ontogenetic changes in foraging ecology and habitat use in long-lived, cryptic marine species

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

Appendix B. Life history pattern classification method.

Life history pattern classification method

Appendix A. Turtle and prey stable isotope analyses.

Turtle and prey stable isotope analyses

Age and residency duration of loggerhead turtles at a North Pacific bycatch hotspot using skeletochronology.

For migratory marine animals, like sea turtles, effective conservation can be challenging because key demographic information such as duration of life stages and exposure to spatially explicit threats in different habitats are often unknown. In the eastern Pacific near the Baja California Peninsula (BCP), Mexico, tens of thousands of endangered North Pacific loggerhead sea turtles (Caretta caretta) concentrate at a foraging area known to have high rates of fishery bycatch. Because stage survivorship of loggerheads in the BCP will vary significantly depending on the number of years spent in this region, we applied skeletochronology to empirically estimate residency duration in this loggerhead hotspot. The observed age distribution obtained from skeletochronology analysis of 146 dead-stranded loggerheads ranged from three to 24 years old, suggesting a BCP residency of >20 years. Given the maximum estimated age and a one-year migration to western Pacific nesting beaches, we infer age-at-maturation for BCP loggerheads at ~25 years old. We also examine survivorship at varying BCP residency durations by applying our findings to current annual mortality estimates. Predicted survivorship of loggerheads spending over 20 years in this BCP foraging habitat is less than 10%, and given that ~43,000 loggerhead turtles forage here, a significant number of turtles are at extreme risk in this region. This is the first empirical evidence supporting estimated age-at-maturation for BCP North Pacific loggerheads, and the first estimates of BCP stage survivorship. Our findings emphasize the urgent need for continued and effective international conservation efforts to minimize bycatch of this endangered species

Female-Bias in a Long-Term Study of a Species with Temperature-Dependent Sex Determination: Monitoring Sex Ratios for Climate Change Research

<div><p>Alterations have occurred and continue to manifest in the Earth’s biota as a result of climate change. Animals exhibiting temperature dependent sex determination (TSD), including sea turtles, are perhaps most vulnerable to a warming of the Earth as highly skewed sex ratios can result, potentially leading to population extinction resulting from decreased male recruitment. Recent studies have begun to quantify climate change impacts to sea turtle populations, especially in terms of predicting effects on hatchling sex ratios. However, given the inherent difficulty in studying sex ratios at this life stage, a more accurate assessment of changes in population sex ratios might be derived by evaluating the juvenile portion of foraging aggregations. We investigated the long-term trend in sex ratio of a juvenile loggerhead (<i>Caretta caretta</i>) sea turtle population inhabiting Pamlico and Core Sounds, North Carolina, USA. We used plasma testosterone reference ranges measured using radioimmunoassay (RIA) to assign sex for 959 turtles and confirmed sex assignment of a subset (N = 58) of the sampled turtles through laparoscopic examination of their gonads. Our results demonstrate that for this particular population of loggerheads, sex ratios (3Females:1Male) had not significantly changed over a 10 year period (1998–2007), nor showed any significant difference among 5-cm straight carapace length (SCL) size classes. Ultimately, these findings provide a basis for comparison with future sex ratios, and highlight the importance of establishing similar long-term studies monitoring secondary, rather than primary, sex ratios, so that needed mitigation measures to climate change impacts can be implemented.</p></div

Age and residency duration of loggerhead turtles at a North Pacific bycatch hotspot using skeletochronology.

For migratory marine animals, like sea turtles, effective conservation can be challenging because key demographic information such as duration of life stages and exposure to spatially explicit threats in different habitats are often unknown. In the eastern Pacific near the Baja California Peninsula (BCP), Mexico, tens of thousands of endangered North Pacific loggerhead sea turtles (Caretta caretta) concentrate at a foraging area known to have high rates of fishery bycatch. Because stage survivorship of loggerheads in the BCP will vary significantly depending on the number of years spent in this region, we applied skeletochronology to empirically estimate residency duration in this loggerhead hotspot. The observed age distribution obtained from skeletochronology analysis of 146 dead-stranded loggerheads ranged from three to 24 years old, suggesting a BCP residency of &gt;20 years. Given the maximum estimated age and a one-year migration to western Pacific nesting beaches, we infer age-at-maturation for BCP loggerheads at ~25 years old. We also examine survivorship at varying BCP residency durations by applying our findings to current annual mortality estimates. Predicted survivorship of loggerheads spending over 20 years in this BCP foraging habitat is less than 10%, and given that ~43,000 loggerhead turtles forage here, a significant number of turtles are at extreme risk in this region. This is the first empirical evidence supporting estimated age-at-maturation for BCP North Pacific loggerheads, and the first estimates of BCP stage survivorship. Our findings emphasize the urgent need for continued and effective international conservation efforts to minimize bycatch of this endangered species