Male Hatchlings Research Articles

Low‐cost tools mitigate climate change during reproduction in an endangered marine ectotherm

Abstract The impacts of anthropogenic climate change will be most dramatic for species that live in narrow thermal niches, such as reptiles. Given the imminent threat to biodiversity, and that actions to reduce carbon emissions are not yet sufficient, it is important that a sound evidence base of potential mitigation options is available for conservation managers. Successful incubation and production of male sea turtle hatchlings is threatened by increased global temperatures (sex is determined by the temperature at which eggs incubate). Here we test two conservation tools to reduce incubation temperatures: clutch splitting and clutch shading, on a nesting loggerhead turtle Caretta caretta population in the Eastern Atlantic Ocean. During the thermosensitive period of incubation, split and shaded clutches were both 1.00°C cooler than control nests. Clutch splitting (mean: 45 eggs) reduced nest temperatures by reducing metabolic heating during incubation compared to controls (mean: 92 eggs). Modelled primary sex ratios differed between nest treatments, with 1.50% (±6% SE) females produced in shaded nests, 45.00% (±7% SE) females in split nests and 69.00% (±6% SE) females in controls. Neither treatment affected hatchling size, success, mass or vigour. When clutch splitting was repeated 2 years later, hatch success was higher in split clutches compared to controls. Synthesis and applications. Clutch splitting and clutch shading successfully altered the thermal profile of incubating turtle nests. When there is sufficient knowledge to better understand the effects of intervention on fundamental population demographics, they will be useful for reducing incubation temperatures in sea turtle nests, potentially increasing nest survival and male hatchling production. The effect of clutch splitting in reducing nest temperature was lower relative to clutch shading, but requires significantly less funding, materials and specialist skill, key factors for management of turtle rookeries that are often in remote, resource‐limited areas.

Artificial and natural shade: Implications for green turtle (Chelonia mydas) rookery management

Global sea turtle populations are threatened by rising atmospheric temperatures which result in higher incubation temperatures. Increased incubation temperatures affect embryo survival, hatchling sex ratio and hatchling locomotor performance. This study aimed to determine the effectiveness of natural and artificial shade on reducing green turtle (Chelonia mydas) incubation temperature at Chagar Hutang beach, Redang Island, Malaysia. To achieve this objective, clutches were incubated using three treatments: unshaded control, artificial shade, and natural tree shade. Estimated hatchling sex ratios, hatching success and locomotor performance were compared across the shade treatments. Clutches incubated in the shaded hatchery were on average 1.3 °C cooler than the unshaded control group, but remained above 30.5 °C and were predicted to produce 100% female hatchlings. In contrast, natural shade from the fringing forest reduced incubation temperatures by 2.7 °C relative to the unshaded clutches, and were predicted to produce up to 85% male hatchlings. Hatching success was similar across treatments and little effect of treatment was observed on hatchling locomotor performance. We recommend relocating nests at high risk of inundation or predation to natural tree shade as a viable solution for ensuring at least some male hatchling production at Chagar Hutang beach.

Research Note: Effects of the in ovo injection of organic zinc, manganese, and copper and posthatch holding time before placement on broiler body temperature during grow out

Effects of the in ovo injection of organic microminerals (OM) (zinc, manganese, and copper) and posthatch holding time (HT) on the daily body temperature (bt) of broilers during grow out were determined. The hatching eggs from a Ross 708 breeder flock at 32 wk of age were incubated under standard commercial conditions. At 17 d of incubation, eggs were randomly allocated to 3 in ovo OM injection treatment (TRT) groups, and at 21 d of incubation, male hatchlings were randomly allocated to 2 posthatch HT treatment groups. Eggs were either not injected or were in ovo injected with diluent only or diluent containing the OM mixture. A 0-hour HT group had immediate access to water and feed, and a 24-hour HT (24HT) group contained birds that were kept in transport baskets in their pens without access to feed and water for 24 h before being released. Fifteen male birds were placed in each of 36 litter floor pens in a temperature-controlled facility. Approximately 2 birds in each of 6 replicate pens belonging to each TRT-HT combination had temperature transponders inserted subcutaneous in the mid-dorsal region of the neck. All birds were brooded under standard commercial conditions and had ad libitum access to feed and water after their respective HT. The bt of the same birds were determined daily at the same time each day beginning at hatch and ending on 39 d of posthatch age (AGE). There were no significant main or interactive effects involving TRT or HT for bt. However, there was a significant (P ≤ 0.0001) main effect because of AGE. A general increase in bt occurred during the 39 d grow out period. At hatch, bt was 40.54 ± 0.056°C and at AGE 39 was 41.46 ± 0.055°C. Under standard brooding conditions, a general increase in bt occurred in the Ross 708 broilers. However, these birds did not exhibit a significant bt response to TRT or a 24HT before placement.

Low male production at the world’s largest green turtle rookery

Because the sex of all sea turtle hatchlings is determined by incubation temperature, with low temperatures producing mainly males and high temperatures producing mainly females, sea turtle populations worldwide are threatened by feminization of hatchlings due to increases in global temperature. Data obtained by laparoscopic sexing of immature individuals captured from a major feeding ground indicates that over several decades there has been little recruitment of males into the northern Great Barrier Reef (nGBR) green turtle Chelonia mydas population, one of the largest sea turtle populations in the world. Over 2 nesting seasons, we measured nest temperatures at Raine Island, the most important nesting site for this nGBR population, and predicted that almost all nests would have produced all female hatchlings. The few nests that produced some male hatchlings were constructed at the very end of the nesting season, and these nests had the lowest hatching success. Taking into account monthly variations in nest construction, hatching success, and hatchling sex ratio, we estimate that over an entire nesting season only 0.7% of hatchlings produced are male. Hence, we conclude that the nGBR population of green turtles has likely recruited very few males in recent years.

Male-Biased Sex Ratio of Adult-Sized Loggerhead Turtles (Caretta caretta) Migrating to High-Latitude Summer-Restricted Foraging Grounds in the Northwest Pacific1

Although part of the life history of loggerhead turtles (Caretta caretta) in the northwest Pacific population has been documented, there is still a paucity of data regarding their spatiotemporal ecology at the local level. The Sanriku Coast is known to be the high-latitude foraging grounds (39° N) for loggerhead turtles of the northwest Pacific population. This site is located far north from the northernmost loggerhead rookeries (>400 km) and more than 10 years of surveys have found that loggerhead turtles migrate there only in the summer. To better understand the spatiotemporal ecology of this population along the Sanriku Coast, the present study investigated the sex of 164 loggerhead turtles captured as bycatch along the Sanriku Coast between 2013 and 2017 by measuring the external morphology and serum testosterone concentrations. These analyses suggested that most of the adult-sized turtles in the Sanriku foraging grounds were males (few females), whereas both sexes of immature-sized turtles migrated to this Coast. This is possibly because migration to the Sanriku Coast is restricted to summer due to the seasonal sea surface temperature shift, and this period overlaps with the nesting period. Therefore, only immature turtles and adult males may be able to migrate. Understanding sex differences in spatiotemporal ecology should contribute to conservation practice in this population, where global warming, together with the turtle's temperature-dependent sex determination, can be expected to reduce the proportion of male hatchlings in the future.

Endocrine and physiological responses of hatchling American kestrels (Falco sparverius) following embryonic exposure to technical short-chain chlorinated paraffins (C10-13)

Short chain chlorinated paraffins (SCCPs) are complex mixtures of polychlorinated n-alkanes, shown to bioaccumulate but with unknown effects in wild birds. The present study examined development-related effects of SCCPs on captive American kestrels (Falco sparverius) treated in ovo on embryonic day (ED) 5 by injection with technical Chloroparaffin® (C10-13, 55.5% Cl) at environmentally relevant nominal (measured) concentrations of 10 (10), 50 (29) or 100 (97) ng ΣSCCP/g egg ww, and artificially incubated until hatching (ED27–ED29). The SCCP concentrations measured in the yolk sacs of the hatchling kestrels bracketed concentrations reported in the eggs of wild birds. Uptake and deposition of these SCCPs differed between male and female hatchlings, with only males showing differences in SCCP concentrations, being highest in the high-dose males than each of the other male groups. Embryonic exposure to SCCPs suppressed glandular total thyroxine (TT4) (20–33%) and reduced circulating triiodothyronine (TT3) (37–40%) in male hatchlings only when compared to control males, but had no effect on glandular TT3 or circulating TT4 in male or female kestrels. Histological assessments of thyroid glands showed that both sexes experienced significant structural changes indicative of gland activation. These thyroid glandular changes and the variations in SCCP concentrations were related to circulating TT3 in female hatchlings. Hepatic deiodinase enzyme (D1, D2) activities were stable and no SCCP-related changes were observed in hatching success, hatchling size, or immune organ size. However, several of the thyroid function indicators were correlated with hatchling size and smaller bursas and spleens, possibly indirectly through SCCP-induced changes in thyroid function. Because changes in thyroid function were evident at concentrations measured in wild bird eggs, similar changes may occur in wild nestlings. The potential impact of these changes on thyroid-mediated growth and survival in wild birds requires further investigation.

Production of male hatchlings at a remote South Pacific green sea turtle rookery: conservation implications in a female-dominated world

Sea turtles have temperature-dependent sex determination, with males being produced at low incubation temperatures and females at high temperatures within the thermal range for embryonic development. In the context of climate change, there are concerns that warming temperatures will lead to an increase in female production. If primary sex ratios are extremely skewed, low male production (and subsequent male availability) may threaten long-term population viability. Heightening these concerns is the fact that female-biased sex ratios are already reported at the majority of sea turtle nesting sites across the world. Here, we describe for the first time primary sex ratios at a remote green sea turtle (Chelonia mydas) nesting site in the South Pacific. Nesting surveys were conducted on Tetiaroa, French Polynesia, between the 2007/2008 and 2018/2019 nesting seasons and revealed a trend of increasing annual nest numbers with large inter-annual fluctuations. We deployed temperature loggers to record incubation conditions and estimated hatchling sex ratios. We recorded low incubation temperatures (mean = 28.5 °C, standard deviation = 0.7 °C) and estimated that currently 54% of all hatchlings produced are male. Low incubation temperatures may be linked to light sand color, shading from vegetation behind beaches and heavy rainfall. Since this site will likely continue to produce males in the future, there is reason for cautious optimism for this population of green sea turtles.

Effects of in ovo injection of L-ascorbic acid on growth performance, carcass composition, plasma antioxidant capacity, and meat quality in broiler chickens1,2,3.

The current study was designed to investigate the effects of the in ovo injection of different dosages of L-ascorbic acid (AA) on posthatch growth performance, carcass characteristics, plasma antioxidant capacity, and meat quality in broiler chickens. A total of 2,220 Ross 708 broiler hatching eggs containing live embryos at 17 D of incubation were subjected to 1 of 6 treatments (trt): non-injected control, saline-injected control, or saline containing 3, 6, 12 or 36mg of AA. An Inovoject m semi-automatic multi-egg injector was used to inject a 100 μL volume of sterile saline (0.85%) alone or containing the different AA dosage into each egg. After hatch, 14 male hatchlings from each trt group were randomly selected and placed in each of 10 replicate floor pens for growth performance evaluation through 45 D posthatch. Chicks from the 3 and 6mg AA trt groups had a higher average daily BW gain and a higher feed intake compared to the saline-injected control group during the grower phase. Chicks in the 12mg AA trt group exhibited a better feed efficiency in the late finisher phase and the entire growing phase than those in the saline-injected control group. Higher thigh and leg percentages were observed in chicks from the 3 and 6mg AA trt groups than in the non-injected control group. Compared to the non-injected or saline-injected control groups, birds in the 12mg AA trt group were observed to have a lower plasma malondialdehyde content during the grower and finisher phases, and breast muscle tenderness was elevated in chicks from the 36mg AA trt group. Taken together, these results suggest that the in ovo injection of AA (3 to 12mg per egg) have lasting positive effects on the posthatch growth, leg muscle development, and systemic antioxidant capacity of broilers. Higher injected dosages of AA (36mg per egg) may also have the potential to improve broiler meat quality.

Equitable Chick Survival in Three Species of the Non-Migratory Shorebird Despite Species-Specific Sexual Dimorphism of the Young.

Simple SummaryWe aimed to determine whether seasonal brood sex-ratio, sex-biased chick survival, and sex specific dimorphism at hatching or during growth occurs among three species of resident Australian shorebird. Our results describe no sex-bias in chick production, survival or growth rates between sexes for any of the three species studied. Sex-biases in populations can have important implications for species’ social biology, population demography and mating systems. It has recently been suggested that in some shorebirds, sex-specific bias in survival of precocial young may occur. This may be driven by variation in the brood sex-ratio and/or the sexual size dimorphism of young birds, which may influence predator escape capacity. Understanding the survival of young birds remains a significant knowledge gap for many taxa, especially when young birds are mobile and cryptic. Our aims were to estimate the sex-ratio variation in three species of Australian resident shorebird, specifically to determine: (1) whether seasonal brood sex-ratio variation at hatching is occurring, (2) the extent of any sex-biased chick survival, (3) if sex specific dimorphism at hatching or during growth occurs; and, (4) whether escape capacity differs between the sexes. We radio-tracked 50 Masked Lapwing Vanellus miles, 42 Red-capped Plover Charadrius ruficapillus and 27 Hooded Plover Thinornis cucullatus chicks from individual broods, examined the likelihood of hatchlings being male or female based on the hatching date within the breeding season, and compared size at hatching, growth and mortality of chicks of different sexes. There was no sex-bias with the hatching date across the breeding season, nor were there differences in survival or growth rates between sexes for any of the three species studied. In one species, male hatchlings had longer tarsi than females, but this did not result in differential escape propensity or improved survival. In conclusion, the hatching date, survival and growth of chicks from three species of resident shorebird was not influenced by their sex.

Effect of incubation temperature on sex-dependent embryo mortality and morphological traits in Mallard.

Although birds have genetically determined sex, the sex ratio has been reported to deviate from parity in several studies. Temperature-dependent sex determination, which is common in reptiles, is absent in birds. However, females are able to adjust their investment into eggs according to the sex of the embryo, which may cause sex-specific embryonic mortality. Incubation temperature may also cause sex-biased embryonic mortality, and it may differentially affect the phenotype of male and female hatchlings. We aimed to investigate differences between male and female Mallard embryos regarding their egg size, mortality during incubation and hatchling phenotype in relation to incubation temperature. Mallard eggs were incubated under six constant incubation temperatures (ranging from 35.0 to 38.0 °C). Hatchlings were weighed, and their morphological traits were measured. We determined the sex of hatchlings and unhatched embryos by genetic analysis and found higher male embryonic mortality at 35.5 °C (44 males vs. 28 females) and a higher proportion of female hatchlings at 38 °C (24 males vs. 38 females); however, these results were not statistically significant. Our results suggest that Mallard females do not differentiate quantitatively between sexes during egg production. Male hatchlings were significantly larger but not heavier than females. The size difference between sexes was most pronounced at temperatures around 36 °C, which is the mean temperature of naturally incubated Mallard eggs.

In ovo exposure to brominated flame retardants Part II: Assessment of effects of TBBPA-BDBPE and BTBPE on hatching success, morphometric and physiological endpoints in American kestrels

Tetrabromobisphenol A bis(2,3-dibromopropyl ether) (TBBPA-BDBPE) and 1,2-bis(2,4,6-tribromophenoxy)ethane (BTPBE) are both brominated flame retardants (BFRs) that have been detected in birds; however, their potential biological effects are largely unknown. We assessed the effects of embryonic exposure to TBBPA-BDBPE and BTBPE in a model avian predator, the American kestrel (Falco sparverius). Fertile eggs from a captive population of kestrels were injected on embryonic day 5 (ED5) with a vehicle control or one of three doses within the range of concentrations that have been detected in biota (nominal concentrations of 0, 10, 50 or 100 ng/g egg; measured concentrations 0, 3.0, 13.7 or 33.5 ng TBBPA-BDBPE/g egg and 0, 5.3, 26.8 or 58.1 ng BTBPE/g egg). Eggs were artificially incubated until hatching (ED28), at which point blood and tissues were collected to measure morphological and physiological endpoints, including organ somatic indices, circulating and glandular thyroid hormone concentrations, thyroid gland histology, hepatic deiodinase activity, and markers of oxidative stress. Neither compound had any effects on embryo survival through 90% of the incubation period or on hatching success, body mass, organ size, or oxidative stress of hatchlings. There was evidence of sex-specific effects in the thyroid system responses to the BTBPE exposures, with type 2 deiodinase (D2) activity decreasing at higher doses in female, but not in male hatchlings, suggesting that females may be more sensitive to BTBPE. However, there were no effects of TBBPA-BDBPE on the thyroid system in kestrels. For the BTPBE study, a subset of high-dose eggs was collected throughout the incubation period to measure changes in BTBPE concentrations. There was no decrease in BTBPE over the incubation period, suggesting that BTBPE is slowly metabolized by kestrel embryos throughout their ∼28-d development. These two compounds, therefore, do not appear to be particularly toxic to embryos of the American kestrel.

Sex Determination in Green Turtle Hatchlings: Geometric Morphometry and Molecular Sex Markers

Determination of the sex ratio in sea turtle population is one of the most important parameters for
 the conservation and management of the species. We aimed to the test usability of the more than
 one approaches that identify the sex of hatchlings with alternative non-invasive methods.
 Geometric morphometry and molecular sex marker were used to indicate difference between the
 sexes in the green sea turtle (Chelonia mydas) hatchlings which sexes were previously determined
 by gonad histology. A total of 60 landmarks were identified from three body parts (carapace,
 plastron and tail) for geometric morphometry. No sex specific difference was found in these three
 body parts. Furthermore, we analyzed a total of 55 different oligonucleotide combinations using
 sex-based pool strategy but found no difference. These two techniques are insufficient for sex
 determination in the green turtle hatchlings. A relatively small number of the primer combinations
 and male hatchlings used to identify sex of C. mydas appears to be a disadvantage. In order to
 obtain clearer results with geometric morphometric and molecular sex markers, it is recommended
 to compare sexes in laboratory conditions with constant temperature.

Climate change resilience of a globally important sea turtle nesting population.

Few studies have looked into climate change resilience of populations of wild animals. We use a model higher vertebrate, the green sea turtle, as its life history is fundamentally affected by climatic conditions, including temperature-dependent sex determination and obligate use of beaches subject to sea level rise (SLR). We use empirical data from a globally important population in West Africa to assess resistance to climate change within a quantitative framework. We project 200years of primary sex ratios (1900-2100) and create a digital elevation model of the nesting beach to estimate impacts of projected SLR. Primary sex ratio is currently almost balanced, with 52% of hatchlings produced being female. Under IPCC models, we predict: (a) an increase in the proportion of females by 2100 to 76%-93%, but cooler temperatures, both at the end of the nesting season and in shaded areas, will guarantee male hatchling production; (b) IPCC SLR scenarios will lead to 33.4%-43.0% loss of the current nesting area; (c) climate change will contribute to population growth through population feminization, with 32%-64% more nesting females expected by 2120; (d) as incubation temperatures approach lethal levels, however, the population will cease growing and start to decline. Taken together with other factors (degree of foraging plasticity, rookery size and trajectory, and prevailing threats), this nesting population should resist climate change until 2100, and the availability of spatial and temporal microrefugia indicates potential for resilience to predicted impacts, through the evolution of nest site selection or changes in nesting phenology. This represents the most comprehensive assessment to date of climate change resilience of a marine reptile using the most up-to-date IPCC models, appraising the impacts of temperature and SLR, integrated with additional ecological and demographic parameters. We suggest this as a framework for other populations, species and taxa.

Sex-specific responses in neuroanatomy of hatchling American kestrels in response to embryonic exposure to the flame retardants bis(2-ethylhexyl)-2,3,4,5-tetrabromophthalate and 2-ethylhexyl-2,3,4,5-tetrabromobenzoate.

Bis(2-ethylhexyl)-2,3,4,5-tetrabromophthalate (BEH-TEBP) and 2-ethylhexyl-2,3,4,5-tetrabromobenzoate (EH-TBB), flame retardant components of FireMaster 550® and 600® have been detected in tissues of wild birds. To address the paucity of information regarding potential impacts of flame retardants on the brain, brain volume regions of hatchling American kestrels (Falco sparverius) were evaluated following in ovo injection at embryonic day 5 with safflower oil or to 1 of 3 doses of either BEH-TEBP (12, 60, or 107 ng/g egg) or EH-TBB (11, 55, or 137 ng/g egg). The doses for both chemicals reflected concentrations reported in wild birds. The volumes of the hippocampus and telencephalon and volumetric differences between left and right hemispheres were measured in hatchlings (embryonic day 28). A sex-specific effect of BEH-TEBP on relative hippocampus volume was evident: the hippocampus was significantly enlarged in high-dose females compared to control females but smaller in low-dose females than the other females. There was no significant effect of EH-TBB on hippocampus volume in female kestrel hatchlings or of either chemical in male hatchlings and no effects of these concentrations of EH-TBB or BEH-TEBP on telencephalon volume or the level of symmetry between the hemispheres of the brain. In sum, embryonic exposure of female kestrels to these BEH-TEBP concentrations altered hippocampus volume, having the potential to affect spatial memory relating to ecologically relevant behavior such as prey capture, predator avoidance, and migration. Environ Toxicol Chem 2018;37:3032-3040. © 2018 SETAC.

Sex-Specific Effects of Incubation Temperature on Embryonic Development of Zebra Finch (Taeniopygia guttata) Embryos.

In oviparous species, the embryonic environment-particularly temperature-can alter phenotype and survival of an individual by affecting its size as well as its metabolic rate. Previous studies have shown that incubation temperatures can affect sex ratio in birds; specifically, low incubation temperatures were shown to produce a male-biased sex ratio in zebra finches (Taeniopygia guttata) possibly because of a higher pre- or postnatal mortality rate in females. We hypothesized that sexes respond differently to suboptimal incubation temperature, leading to a male-biased sex ratio. To test this hypothesis, zebra finch eggs were incubated at 36.1°, 37.5°, or 38.5°C and hatching success, hatchling mass, residual yolk mass, and pectoralis mass were measured. We found that while hatchling mass was similar between the sexes at 37.5°C, female hatchlings were heavier at 36.1°C, and male hatchlings were heavier at 38.5°C. Pectoralis muscle mass was similar between the sexes at 36.1°C; however, at 37.5°C, female pectoralis mass was heavier at hatching than that of males. Females at 37.5°C also had lower residual yolk at hatching compared with males, reflecting a higher use of energy by female embryos compared with male embryos at this temperature. In contrast, residual yolk was similar between the sexes at 36.1° and 38.5°C. Our results suggest that there are sex differences in how incubation temperature alters organ mass and yolk energy reserve; this can lead to a difference in survival at different incubation temperatures between the sexes. Taken together with previous studies showing that females alter incubation behavior with ambient temperature, rising ambient temperatures could impact phenotype and survival of avian offspring in a sex-specific manner.

Going back to the roots: finding a strategy for the management of nesting loggerhead sea turtles in New Caledonia

Incubation temperature plays a vital role in sea turtle life history because it influences embryonic growth, sex determination and hatchling attributes such as body size, residual yolk size, self-righting ability, crawling speed and swimming speed. For these reasons there is concern that predicted increases in air temperature, as a result of global warming, will increase nest temperatures and result in decreased hatching success, decrease or cease male hatchling production, and decreased hatchling quality. In a previous study examining incubation temperature at a loggerhead turtle (Caretta caretta) rookery located at La Roche Percée, New Caledonia, high nest temperatures and root invasion by beach morning glory (Ipomoea pes-caprae) were found to adversely affect hatching success and locomotor performance. In the current study, we relocated loggerhead turtle nests into shaded hatcheries. Shading nests decreased sand and nest temperatures and was predicted to increase male hatchling production slightly, but nest emergence success was decreased due to invasion of cottonwood (Hibiscus tiliaceus) roots into some nests. Using shaded structures is a viable and affordable management option to counteract the high sand temperatures found on some sea turtle nesting beaches, but these shade structures need to be located some distance from trees and other plants to ensure that root penetration into nests does not adversely affect nest emergence success.

Impacts of Invasive Phragmites australis on Diamondback Terrapin Nesting in Chesapeake Bay

The range expansion of exotic plant species, including the invasive reed Phragmites australis, causes widespread structural and functional changes to coastal ecosystems along the Atlantic and Gulf Coasts of North America. Native estuarine species, such as the diamondback terrapin (Malaclemys terrapin), are at risk of adverse effects from rapid habitat changes due to exotic invasions. Diamondback terrapins currently face population threats including by-catch mortality in crab pots, predation, and habitat loss, and populations may continue to suffer if deleterious plant invasions into preferred nesting habitats are left unchecked. We examined the extent to which Phragmites affects nesting of a breeding population of diamondback terrapins at Fisherman Island National Wildlife Refuge on the eastern shore of Virginia, where Phragmites has recently expanded into known areas of terrapin nesting. With data collected from the 2015 nesting season, we quantified the extent to which Phragmites shading could impact nest incubation temperature and determined how Phragmites density impacts the risk of rhizome invasion into nests. We conclude that Phragmites cover greater than 50% would decrease incubation temperatures of terrapin nests sufficiently to produce predominantly male hatchlings. Phragmites cover had no observed effect on root growth into simulated nests, but cover by other dune plant species explained observed trends in root growth. These results suggest that terrapins may be negatively impacted by Phragmites expansion into open nesting sites. Breeding site fidelity exhibited by terrapins and other estuarine species could limit the ability of their populations to adjust to rapid coastal expansion of invasive plant species.

Assessment of hatchery management for the loggerhead turtle (Caretta caretta) nests on Göksu Delta, Turkey

Sea turtles suffer from numerous threats such as predation, tidal inundation, and coastal building. We aimed to assess the effectiveness of hatchery technique on Göksu Delta beach, Turkey during the nesting season of 2010. For this purpose, eggs of loggerhead turtle (Caretta caretta) nests under the risk of tidal inundation were transferred into two protected hatchery sites that were constructed on Göksu Delta beach, in which there were no risk of predation and inundation, and the other nests were left to remain in situ. Hatching success of hatchery and natural nests were calculated. In addition, temperatures of both hatchery and natural nests (5 and 6 nests, respectively) were recorded, and sex ratios were determined using these nest temperatures and incubation periods. It was determined that hatching success increased with increasing distance from the sea (until 31 m) (r2 = 0.98), increasing incubation period (up to 60 days) (r2 = 0.68), and decreasing clutch size (r2 = 0.93), implying that the production of male hatchling is high in a nest if hatching success of that nest is high (or vice versa). Mean hatching success, incubation period, and nest temperature for hatchery nests were found to be 72.8%, 51.1 days, and 30.2 °C, respectively, while the corresponding values for natural nests were found to be 40.5%, 49.9 days, and 31.1 °C, respectively. It was also detected that hatchery nests produced higher proportion of male hatchlings (25.2%) compared with natural nests (13.5%). Based on all these results, it can be concluded that hatchery management on this beach is not a completely effective conservation technique. Nest relocation and/or hatchery management enable the conservationists to increase hatching success rate, but they cause sex ratio alteration. In this respect, we can only partially support nest relocation and/or hatchery management. They should be considered as a last option due to sex ratio alteration.

The effects of nest incubation temperature on embryos and hatchlings of the loggerhead sea turtle: Implications of sex difference for survival rates during early life stages

In sea turtles, higher hatch and emergence successes, larger body size of hatchlings, and superior terrestrial and swimming locomotor performances are important to maximize survival rates. Here, the hatch and emergence successes, embryonic mortality rates, hatchling morphology, and terrestrial and swimming locomotor performances of the loggerhead sea turtle (Caretta caretta) from male and female producing nests were investigated. Mortality rates during the early embryonic stage in the male producing nests were significantly lower than those in the female producing nests. The hatchlings from male producing nests showed higher terrestrial locomotor performances than those from female producing nests. Furthermore, while the other factors did not show differences between the two nesting groups, the high incubation temperatures significantly affected the mortality rate of embryos and the swimming performance of hatchlings. The present study indicates that male hatchlings are likely to have higher survival rates than female hatchlings during the critical terrestrial period. In addition, the present study suggests that as global warming proceeds, female-fate embryos and/or female hatchlings are likely to face lower survival rates than males, owing to high mortality rate of embryos and low swimming performance of hatchlings.

Histochemical and immunohistochemical studies of the gonads and paramesonephric ducts of male and female hatchlings of loggerhead sea turtles (Caretta caretta)

The sex of neonatal sea turtles is difficult to determine, because neonates lack heteromorphic sex chromosomes and dimorphic external characteristics; internal dimorphic morphology is defined at hatching. We used histochemical staining and made measurements in the gonads and paramesonephric ducts (PD) of both sexes to determine structural differences in female and male loggerhead sea turtle (Caretta caretta) hatchlings. We detected differences in the gonads and PD between the sexes including the amounts of mucopolysaccharides, collagen and elastic fibers. We determined that the thickness of the gonadal cortex and the diameter of the PD lumen are reliable sex-specific characteristics. We also assessed immunolocalization of aromatase, an enzyme complex that converts androgens to estrogens, and found differences in the localization and intensity of aromatase immunostaining in the gonads and PD of female and male hatchlings. Comprehensive studies of the sexual differences of sea turtles are important for conservation programs.