Eradication of Salmonella and Arizona species from turtle hatchlings produced from eggs treated on commercial turtle farms.

On commercial turtle farms more than 40% of the hatchlings excrete detectable levels of Salmonella and Arizona spp. when hatched from nonsanitized eggs incubated in sawdust or dirt-filled chambers. Over a 3-year period on 10 farms, more than 10(6) turtle eggs were treated in an attempt to hatch Salmonella-free turtles. Eggs were sanitized in disinfectant, treated by temperature- or pressure-differential dip methods in solutions containing 500 micrograms or more of gentamicin sulfate per ml, and hatched in sanitized plastic chambers free of bedding material. The Salmonella and Arizona spp. infection levels for turtles produced from treated eggs were 0 and 1.12% for years 1 and 2, respectively, whereas infection levels for hatchlings produced from nontreated eggs during these periods were 47 and 44%, respectively. During year 3, dip solutions were filtered daily, treated at 100 degrees C for 15 min on a weekly basis to free the solution of microbial contaminants and egg protein, charged with gentamicin after 10,000 to 20,000 eggs had been treated to maintain antimicrobial activity at 500 micrograms/ml or more, and maintained at pH 6.0 to preserve optimal antimicrobial activity. The implementation of these measures in year 3 resulted in an infection level of 0.15% when the tissues of 3 of 1,959 hatchlings tested were positive for Salmonella and Arizona spp., whereas the tissues of 66 (49.0%) of 135 hatchlings produced from nontreated eggs were positive.     

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.     

Treatment of Salmonella-Arizona-Infected Turtle Eggs with Terramycin and Chloromycetin by the Temperature Differential Egg Dip Method

Attempts to eliminate Salmonella and Arizona infection from newly hatched turtles were made by dipping fresh eggs in cold solutions of Terramycin and Chloromycetin at 1,000, 1,200, 1,500 and 2,000 mug per ml for either 10, 20, or 30 min. Control groups consisted of hatchings produced from nondipped eggs or eggs dipped in chilled water. In two of the four experiments 5 to 10 eggs were blended on days 15, 30, and 45 post antibiotic dip treatment. Twenty-five to 60 hatchlings from each control or experimental dip groups were held in containers and the water was tested (excretion method) for Salmonella and Arizona every 15 or 30 days for 180 to 210 days after hatching. Representative turtles were homogenized (blending method) to determine if systemic infections were present. All specimens tested were enriched in tetrathionate and selenite cystine broth. Nondipped eggs and water-dipped eggs routinely showed Salmonella and Arizona present in egg homogenate and hatchlings emerging from these eggs excreted these pathogens. Terramycin- and Chloromycetin-dipped eggs were uniformly negative for these pathogens, only if fresh eggs were dipped. Bacteriological assay of container water and whole turtle homogenate from hatchlings were negative for Salmonella and Arizona if eggs were dipped in 1,000 mug of Terramycin early in the egg laying season or if eggs were dipped in 1,500 or 2,000 mug of Terramycin per ml late in the egg laying season. The results of temperature-differential egg dip studies suggest that this is a feasible and promising method by which to eradicate Salmonella and Arizona from the turtle.     

Sea turtle species vary in their susceptibility to tropical cyclones

Severe climatic events affect all species, but there is little quantitative knowledge of how sympatric species react to such situations. We compared the reproductive seasonality of sea turtles that nest sympatrically with their vulnerability to tropical cyclones (in this study, “tropical cyclone” refers to tropical storms and hurricanes), which are increasing in severity due to changes in global climate. Storm surges significantly decreased reproductive output by lowering the number of nests that hatched and the number of hatchlings that emerged from nests, but the severity of this effect varied by species. Leatherback turtles (Dermochelys coriacea) began nesting earliest and most offspring hatched before the tropical cyclone season arrived, resulting in little negative effect. Loggerhead turtles (Caretta caretta) nested intermediately, and only nests laid late in the season were inundated with seawater during storm surges. Green turtles (Chelonia mydas) nested last, and their entire nesting season occurred during the tropical cyclone season; this resulted in a majority (79%) of green turtle nests incubating in September, when tropical cyclones are most likely to occur. Since this timing overlaps considerably with the tropical cyclone season, the developing eggs and nests are extremely vulnerable to storm surges. Increases in the severity of tropical cyclones may cause green turtle nesting success to worsen in the future. However, published literature suggests that loggerhead turtles are nesting earlier in the season and shortening their nesting seasons in response to increasing sea surface temperatures caused by global climate change. This may cause loggerhead reproductive success to improve in the future because more nests will hatch before the onset of tropical cyclones. Our data clearly indicate that sympatric species using the same resources are affected differently by tropical cyclones due to slight variations in the seasonal timing of nesting, a key life history process.     

First fossil gravid turtle provides insight into the evolution of reproductive traits in turtles

Here we report on the first discovery of shelled eggs inside the body cavity of a fossil turtle and on an isolated egg clutch, both referable to the Cretaceous turtle Adocus. These discoveries provide a unique opportunity to gain insight into the reproductive traits of an extinct turtle and to understand the evolution of such traits among living turtles. The gravid adult and egg clutch indicate that Adocus laid large clutches of rigid-shelled spherical eggs and established their nests near rivers, traits that are shared by its closest living relatives, the soft-shelled turtles. Adocus eggshell, however, was probably more rigid than that of living turtles, based on its great thickness and structure, features that may represent unique adaptations to intense predation or to arid nest environments. In light of the reproductive traits observed in Adocus, the distribution of reproductive traits among turtles reveals that large clutches of rigid-shelled eggs are primitive for hidden-necked turtles (cryptodirans) and that spherical eggs may have evolved independently within this group.     

Eradication of Arizona hinshawii from artificially infected turtle eggs.

Turtle eggs, 24 h old, were infected with Arizona hinshawii and treated 48 h later with gentamicin sulfate (Garasol; Shering Corp., Allantown, N.J.) by pressure differential egg dip treatment to ascertain the concentration of this reagent required to eradicate this pathogen from eggs. Infected eggs treated with 1,000 or 1,500 micrograms of gentamicin per ml of dip solution eliminated detectable A. hinshawii from eggs as determined by testing shells and embryo-yolk homogenates of 12-day-old eggs and the gastrointestinal tracts, kidneys, livers and gall bladders, and yolks of 50-day-old embryos. Treated eggs produced hatchlings which did not excrete detectable A. hinshawii at 72 h or 30 days after hatching, nor was this organism recovered from the visceral organs of these hatchlings when necropsied 30 days after hatching. Bacteriological assays on infected nontreated eggs showed that greater than 70% of the eggs harbored A. hinshawii, and eggs in this group produced hatchlings which actively excreted and harbored A. hinshawii. Eggs not infected or treated also produced turtles which excreted and systemically carried A. hinshawii and Salmonella spp. though not at the same level as did the turtles produced from infected, nontreated eggs.     

Eradication of Arizona hinshawii from artificially infected turtle eggs

Turtle eggs, 24 h old, were infected with Arizona hinshawii and treated 48 h later with gentamicin sulfate (Garasol; Shering Corp., Allantown, N.J.) by pressure differential egg dip treatment to ascertain the concentration of this reagent required to eradicate this pathogen from eggs. Infected eggs treated with 1,000 or 1,500 micrograms of gentamicin per ml of dip solution eliminated detectable A. hinshawii from eggs as determined by testing shells and embryo-yolk homogenates of 12-day-old eggs and the gastrointestinal tracts, kidneys, livers and gall bladders, and yolks of 50-day-old embryos. Treated eggs produced hatchlings which did not excrete detectable A. hinshawii at 72 h or 30 days after hatching, nor was this organism recovered from the visceral organs of these hatchlings when necropsied 30 days after hatching. Bacteriological assays on infected nontreated eggs showed that greater than 70% of the eggs harbored A. hinshawii, and eggs in this group produced hatchlings which actively excreted and harbored A. hinshawii. Eggs not infected or treated also produced turtles which excreted and systemically carried A. hinshawii and Salmonella spp. though not at the same level as did the turtles produced from infected, nontreated eggs.     

Effects of the egg incubation environment on turtle carapace development

Developing organisms are often exposed to fluctuating environments that destabilize tissue-scale processes and induce abnormal phenotypes. This might be common in species that lay eggs in the external environment and with little parental care, such as many reptiles. In turtles, morphological development has provided striking examples of abnormal phenotypic patterns, though the influence of the environment remains unclear. To this end, we compared fluctuating asymmetry, as a proxy for developmental instability, in turtle hatchlings incubated in controlled laboratory and unstable natural conditions. Wild and laboratory hatchlings featured similar proportions of supernumerary scales (scutes) on the dorsal shell (carapace). Such abnormal scutes likely elevated shape asymmetry, which was highest in natural nests. Moreover, we tested the hypothesis that hot and dry environments cause abnormal scute formation by subjecting eggs to a range of hydric and thermal laboratory incubation regimes. Shape asymmetry was similar in hatchlings incubated at five constant temperatures (26–30°C). A hot (30°C) and severely Dry substrate yielded smaller hatchlings but scutes were not overtly affected. Our study suggests that changing nest environments contribute to fluctuating asymmetry in egg-laying reptiles, while clarifying the conditions at which turtle shell development remains buffered from the external environment.     

Does maternal oviposition site influence offspring dispersal to suitable habitat?

Orientation and dispersal to suitable habitat affects fitness in many animals, but the factors that govern these behaviors are poorly understood. In many turtle species, hatchlings must orient and disperse to suitable aquatic habitat immediately after emergence from subterranean nests. Thus, the location of nest sites relative to aquatic habitats ideally should be associated with the direction of hatchling dispersal. At our study site, painted turtles (Chrysemys picta) nest to the west (on an island) and east (on the mainland) of a wetland, which determines the direction that hatchlings must travel to reach suitable aquatic habitat. To determine if hatchling orientation is intrinsically influenced by the location where their mothers nest, we employed a two-part cross-fostering experiment in the field, whereby half the eggs laid in mainland nests were swapped with half the eggs laid in island nests. Moreover, because C. picta hatchlings overwinter inside their nests, we performed a second cross-fostering experiment to fully decouple the effects of (1) the maternally chosen nest location, (2) the embryonic developmental location, and (3) the overwinter location. We released hatchlings into a circular arena in the field and found that turtles generally dispersed in a westerly direction, regardless of the maternally chosen nest location and independent of the locations of embryonic development and overwintering. Although this westerly direction was towards suitable aquatic habitat, we could not distinguish whether naïve hatchling turtles (i) use environmental cues/stimuli to orient their movement, or (ii) have an intrinsic bias to orient west in the absence of stimuli. Nevertheless, these findings suggest that the orientation behavior of naïve hatchling turtles during terrestrial dispersal is not dependent upon the location of maternally-chosen nest sites.     

Influence of the nest environment on bone mineral content in hatchling painted turtles (Chrysemys picta)

We performed an experiment at a field site in north-central Nebraska to assess the role of the nest environment in inducing variation in bone mineral content in hatchling painted turtles Chrysemys picta (Schneider 1783). The contents of several newly constructed nests were manipulated by reciprocal transplant, after which the eggs were allowed to incubate for 8 wk under natural conditions. The nests were then excavated, and the eggs were brought into the laboratory to complete incubation and hatch under standard conditions of temperature and moisture. The hatchlings were killed, and their carcasses and residual yolks were analyzed separately for calcium and phosphorus. More of the random variation in carcass calcium and phosphorus was related to the nest in which eggs incubated (37% and 42%, respectively) than was associated with the clutch of origin (21% and 37%). Moreover, hatchlings from some nests contained substantially more calcium and phosphorus than did hatchlings from other nests, both in terms of the absolute amounts of the elements in their carcasses (pointing to variation in body size) and in terms of the concentrations of those elements (pointing to variation in bone density). The amounts of calcium and phosphorus in carcasses of hatchlings were positively correlated with changes in mass of their eggs during the 8 wk that the eggs incubated in nests in the field, thereby indicating that the influence of the nest environment on developing embryos probably was mediated by water exchanges experienced by the eggs. These findings indicate that developmental plasticity underlies a major fraction of the variation in mineral content of hatchling painted turtles emerging from nests in the field. Phenotypic variation attributable to plasticity consequently needs to be addressed in models for life-history evolution of painted turtles and other chelonians producing eggs with soft, flexible shells.     

An assessment of initial body size in loggerhead sea turtle (Caretta caretta) hatchlings in Turkey

Eggs, hatchlings, and adult loggerhead turtles, and incubation durations of clutches, were measured on three Turkish beaches (Dalyan, Fethiye and G?ksu Delta), and some physical features of nests were compared. These features were not statistically different among the beaches, except for nest depth and distance to the high water mark. There was a positive relationship between hatchling mass and egg size. The carapace length of hatchlings was correlated with both egg diameter and incubation duration. The duration of asynchronous emergence of hatchlings on Fethiye beach was slightly longer than on the other two beaches, and the size of hatchlings decreased as asynchronous emergence proceeded. Of the hatchlings that emerged first, those that died were significantly smaller in SCL and mass than those that lived. These results suggest that smaller hatchlings may not be vigorous enough to emerge earlier from nests, and that they may be less fit.     

Metabolic heat estimation of the sea turtle Lepidochelys olivacea embryos

Several studies have reported the importance of metabolic heat on the increment of temperature in the sea turtle nests; however, the metabolic heat has not been calculated for sea turtle eggs. In this study, the metabolic heat generated by embryos of the sea turtle Lepidochelys olivacea was estimated from a thermal balance model by means of three measured temperatures—one in the center of the nest, and the others in the sand above and beside the nest. An experiment was conducted with a sample of 100 eggs from a Lepidochelys olivacea nest collected in the Baja Peninsula, Mexico. The results showed that during the incubation period, no metabolic heat was detected before day 19 but it increased from that day until a maximum of 0.84 W at day 34, when the incubation process was interrupted due to rain. This value corresponds to 31 emerged hatchlings. The novel model is a suitable framework to predict the temperature and metabolic heat within the nest.     

A novel hypothesis for the adaptive maintenance of environmental sex determination in a turtle

Temperature-dependent sex determination (TSD) is widespread in reptiles, yet its adaptive significance and mechanisms for its maintenance remain obscure and controversial. Comparative analyses identify an ancient origin of TSD in turtles, crocodiles and tuatara, suggesting that this trait should be advantageous in order to persist. Based on this assumption, researchers primarily, and with minimal success, have employed a model to examine sex-specific variation in hatchling phenotypes and fitness generated by different incubation conditions. The unwavering focus on different incubation conditions may be misplaced at least in the many turtle species in which hatchlings overwinter in the natal nest. If overwintering temperatures differentially affect fitness of male and female hatchlings, TSD might be maintained adaptively by enabling embryos to develop as the sex best suited to those overwintering conditions. We test this novel hypothesis using the painted turtle (Chrysemys picta), a species with TSD in which eggs hatch in late summer and hatchlings remain within nests until the following spring. We used a split-clutch design to expose field-incubated hatchlings to warm and cool overwintering (autumn–winter–spring) regimes in the laboratory and measured metabolic rates, energy use, body size and mortality of male and female hatchlings. While overall mortality rates were low, males exposed to warmer overwintering regimes had significantly higher metabolic rates and used more residual yolk than females, whereas the reverse occurred in the cool temperature regime. Hatchlings from mixed-sex nests exhibited similar sex-specific trends and, crucially, they were less energy efficient and grew less than same-sex hatchlings that originated from single-sex clutches. Such sex- and incubation-specific physiological adaptation to winter temperatures may enhance fitness and even extend the northern range of many species that overwinter terrestrially.     

Environmentally induced variation in body size of turtles hatching in natural nests

Eggs from three snapping turtles (Chelydra serpentina) were divided between two natural nests in a factorial experiment assessing the role of the nest environment as a cause for variation in body size and energy reserves of hatchlings at our study site in northcentral Nebraska. Nest # 1 was located in an unshaded area on the south side of a high sandhill, whereas nest #2 was located in an unshaded area on level ground. Eggs in nest #1 increased in mass over the course of incubation, with eggs at the bottom of the nest gaining more mass than eggs nearer to the surface. In constrast, eggs in nest #2 lost mass during incubation, with eggs at the bottom declining less in mass than eggs at the top of the cavity. Hatchlings from nest #1 were much larger (but contained smaller masses of unused yolk) than hatchlings from nest #2. Additionally, eggs from the lower layers in both nests tended to produce larger hatchings (but with smaller masses of unused yolk) than eggs from the upper layers. Thus, ecologically important variation in body size and nutrient reserves of hatchling snapping turtles results from variation in the environment among and within nests.     

How do goannas find sea turtle nests?

Squamate reptiles rely heavily on visual and chemical cues to detect their prey, so we expected yellow‐spotted goannas (Varanus panoptes) which are predators of sea turtle nests on mainland beaches in northern Australia would use these cues to find sea turtle nests. Ghost crabs (Ocypode ceratophthalmus and Ocypode cordimanus) are also common on Australian sea turtle nesting beaches and frequently burrow into sea turtle nests. However, the potential for ghost crab burrowing activity at sea turtle nests to signal the location of a nest to goannas has not been investigated. Here, we used camera traps and presence of tracks at nests to record goanna activity around selected nests during the incubation period and 10 days after hatchling turtles emerged from their nests. We also recorded the number of ghost crab burrows around nests to evaluate ghost crab activity. Our results indicated that nest discovery by goannas was independent of nest age, but that the nest visitation rate of goannas and crabs increased significantly after a nest had been opened by a goanna or after hatchlings had emerged from the nest. There was no apparent connection between ghost crab burrows into a nest and the likelihood of that nest being predated by goannas.     

Physiological ecology of overwintering in the hatchling painted turtle: multiple-scale variation in response to environmental stress

We integrated field and laboratory studies in an investigation of water balance, energy use, and mechanisms of cold-hardiness in hatchling painted turtles (Chrysemys picta) indigenous to west-central Nebraska (Chrysemys picta bellii) and northern Indiana (Chrysemys picta marginata) during the winters of 1999-2000 and 2000-2001. We examined 184 nests, 80 of which provided the hatchlings (n=580) and/or samples of soil used in laboratory analyses. Whereas winter 1999-2000 was relatively dry and mild, the following winter was wet and cold; serendipitously, the contrast illuminated a marked plasticity in physiological response to environmental stress. Physiological and cold-hardiness responses of turtles also varied between study locales, largely owing to differences in precipitation and edaphics and the lower prevailing and minimum nest temperatures (to -13.2 degrees C) encountered by Nebraska turtles. In Nebraska, winter mortality occurred within 12.5% (1999-2000) and 42.3% (2000-2001) of the sampled nests; no turtles died in the Indiana nests. Laboratory studies of the mechanisms of cold-hardiness used by hatchling C. picta showed that resistance to inoculative freezing and capacity for freeze tolerance increased as winter approached. However, the level of inoculation resistance strongly depended on the physical characteristics of nest soil, as well as its moisture content, which varied seasonally. Risk of inoculative freezing (and mortality) was greatest in midwinter when nest temperatures were lowest and soil moisture and activity of constituent organic ice nuclei were highest. Water balance in overwintering hatchlings was closely linked to dynamics of precipitation and soil moisture, whereas energy use and the size of the energy reserve available to hatchlings in spring depended on the winter thermal regime. Acute chilling resulted in hyperglycemia and hyperlactemia, which persisted throughout winter; this response may be cryoprotective. Some physiological characteristics and cold-hardiness attributes varied between years, between study sites, among nests at the same site, and among siblings sharing nests. Such variation may reflect adaptive phenotypic plasticity, maternal or paternal influence on an individual's response to environmental challenge, or a combination of these factors. Some evidence suggests that life-history traits, such as clutch size and body size, have been shaped by constraints imposed by the harsh winter environment.     

Sea Turtle Nesting as a Process Influencing a Sandy Beach Ecosystem

Sea turtle egg mortality, egg predation, and small organisms associated with turtle nests were studied at Playa Ostional, Costa Rica. Sites with concentrated sea turtle nesting were compared with solitary nesting sites as a function of place and time based on ANOVA, Akaike's Information Criterion, and Bayesian analyses. Results indicate that sea turtle egg mortality was significantly associated (P < 0.005) with flowing water that erodes or saturates nesting sites, and with overlapped nesting in which sea turtles disturb each other's nests. Sarcophagid and calliphorid fly larvae (Bayesian prior = 1.19; posterior = 2.27), fungi (prior = 1.14; posterior = 1.92), mites (prior = 0.51; posterior = 1.15), and several other types of small organisms increased in number after turtle egg laying (N= 303 nests; 34,451 turtle eggs). During peak sea turtle nesting periods, visitation to nesting sites by poachers and vertebrate predators was high, and relative number of nests disturbed by these predators was low (P < 0.02). In multimodel analysis, the three most parsimonious models were: (1) turtle egg mortality and distance from mean high tide; (2) turtle egg predation and distance from mean high tide; and (3) turtle egg mortality and nesting density, with Akaike weights of 0.224, 0.203, and 0.153 respectively. Intensive sea turtle nesting might result in upwelling and turnover of nesting debris and nest organisms, and may influence biotic community structure of sandy beach ecosystems.     

Reproductive Biology of Captive Green Sea Turtles Chelonia mydas

A captive colony of green sea turtles, Chelonia mydas, has beenmaintained and observed at a commercial sea turtle farm on GrandCayman Island, B.W.I., since 1973. Observations of this breedingcolony show that the mating and nesting behaviour of the captivegreen sea turtle is similar to that observed in wild populations.Evidence indicates that mating observed prior to a female'snesting in a given season determines the hatchabilityof thatseason's egg production. Annual per female egg production ofthe captive colony appears to be two to five times greater thanthat reported for wild colonies. Observations on the reproductivebiology of green sea turtles hatched and raised under farm conditionssuggests that the minimum age of sexual maturity is eight tonine years of age. The number of eggs per nest, the number ofnests per season per female and hatch rate tend to increasewith successive seasons nesting for these turtles reaching sexualmaturity.     

Climate change affects the early-life history of a freshwater turtle in a severely drying region

Freshwater turtles are one of the most threatened vertebrate groups. Climate change is a major threat to these species, with impacts affecting all life-history stages. There is currently a limited understanding of how changes in climate may alter the environmental triggers for hatching and emergence from the nests of freshwater turtle hatchlings. This precludes making predictions about how climate change may impact freshwater turtle recruitment success. The southwestern snake-necked turtle (Chelodina oblonga) is endemic to south-western Australia, a global biodiversity hotspot that has undergone severe climatic drying. Recruitment failure is thought to be occurring in many populations of the species. However, there is little understanding as to how environmental change may be influencing recruitment. This study aimed to: (1) determine the incubation duration and hatching and hatchling emergence success of C. oblonga, (2) determine if the species exhibits hatching or emergence synchrony and/or delayed emergence and (3) quantify the effects of temperature and rainfall on hatchling emergence. Using this information, the study assesses how climatic drying and warming may be impacting C. oblonga's early life-history. Between 2018 and 2020 nest sites were monitored around a large urban wetland with weekly assessments of egg and hatchling status. Incubation duration and hatching and hatchling emergence success were calculated, and generalized linear models were built to determine how temperature and/or rainfall predicted emergence. Hatchlings either emerged shortly after hatching or overwintered in the nest, and both hatching and emergence were asynchronous. Both emergence periods were positively associated with temperature and rainfall. This study reveals that incubation duration, hatching success, hatchling emergence and survival are all likely to be impacted by recent and projected climate change, and especially drying. Warming and drying are predicted for many temperate regions globally, and it is therefore important that their impacts on the early life history of freshwater turtles be better understood.