Foxl2 is required for the initiation of the female pathway in a temperature-dependent sex determination system in Trachemys scripta.

KDM6B-mediated epigenetic modification of the testicular regulator Dmrt1 has previously been identified as the primary switch of the male pathway in a temperature-dependent sex-determination (TSD) system; however, the molecular network of the female pathway has not yet been established. Here, we have functionally characterized for the first time an upstream regulator of the female pathway, the forkhead transcription factor FOXL2, in Trachemys scripta, a turtle species with a TSD system. FOXL2 exhibited temperature-dependent female-specific expression patterns before the onset of gonadal differentiation and was preferentially localized in ovarian somatic cells. Foxl2 responded rapidly to temperature shifts and estrogen. Importantly, forced expression of Foxl2 at the male-producing temperature led to male-to-female sex reversal, as evidenced by the formation of an ovary-like structure, and upregulation of the ovarian regulators Cyp19a1 and R-spondin1. Additionally, knockdown of Foxl2 caused masculinization at the female-producing temperature, which was confirmed by loss of the female phenotype, development of seminiferous tubules, and elevated expression of Dmrt1 and Sox9. Collectively, we demonstrate that Foxl2 expression is necessary and sufficient to drive ovarian determination in T. scripta, suggesting a crucial role of Foxl2 in female sex determination in the TSD system.

Systemic Caryospora-like coccidiosis in a clutch of hatchling red-eared slider turtles (Trachemys scripta elegans).

Caryospora-like organisms (CLOs) form a clade of at least 11 genotypes of related coccidia that can cause epizootic mortality in marine turtles. The biology, transmission, host species range, and host cell tropism of these organisms are still largely unknown. The goal of this study was to characterize the host cell tropism, pathologic and ultrastructural features, and phylogeny associated with the first report of a mortality event due to CLO in the freshwater red-eared slider turtle (Trachemys scripta elegans). Sudden mortalities within a clutch of captive-raised red-eared slider hatchlings (n = 8) were recorded, and deceased animals had severe segmental to diffuse, transmural, fibrinonecrotic enterocolitis and multifocal to coalescing hepatic necrosis, among other lesions associated with numerous intracytoplasmic developing stages of intralesional coccidia. Among the different developmental stages, merozoites were ultrastructurally characterized by an apical complex. A pan-apicomplexan polymerase chain reaction (PCR) yielded a 347 bp-amplicon matching the Schellackia/Caryospora-like clade with 99.1% identity to the US3 strain from green sea turtles (Chelonia mydas) and 99.1% identity to Schellackia sp. Isolate OC116. Surviving hatchlings were treated with toltrazuril sulfone (ponazuril) but were subsequently euthanized due to the risk of spreading the parasite to other chelonids in the collection. The ponazuril-treated hatchlings (n = 4) had mild proliferative anterior enteritis, with few intraepithelial coccidia in one hatchling confirmed as CLO by PCR. This is the first report of Caryospora-like coccidiosis in non-cheloniid turtles, highlighting the relevance of this disease as an emerging highly pathogenic intestinal and extra-intestinal form of coccidiosis of turtles with potential cross-species infectivity.

Low production of mitochondrial reactive oxygen species after anoxia and reoxygenation in turtle hearts.

Extremely anoxia-tolerant animals, such as freshwater turtles, survive anoxia and reoxygenation without sustaining tissue damage to their hearts. In contrast, for mammals, the ischemia-reperfusion (IR) injury that leads to tissue damage during a heart attack is initiated by a burst of superoxide (O2·-) production from the mitochondrial respiratory chain upon reperfusion of ischemic tissue. Whether turtles avoid oxidative tissue damage because of an absence of mitochondrial superoxide production upon reoxygenation, or because the turtle heart is particularly protected against this damage, is unclear. Here, we investigated whether there was an increase in mitochondrial O2·- production upon the reoxygenation of anoxic red-eared slider turtle hearts in vivo and in vitro. This was done by measuring the production of H2O2, the dismutation product of O2·-, using the mitochondria-targeted mass-spectrometric probe in vivo MitoB, while in parallel assessing changes in the metabolites driving mitochondrial O2·- production, succinate, ATP and ADP levels during anoxia, and H2O2 consumption and production rates of isolated heart mitochondria. We found that there was no excess production of in vivo H2O2 during 1 h of reoxygenation in turtles after 3 h anoxia at room temperature, suggesting that turtle hearts most likely do not suffer oxidative injury after anoxia because their mitochondria produce no excess O2·- upon reoxygenation. Instead, our data support the conclusion that both the low levels of succinate accumulation and the maintenance of ADP levels in the anoxic turtle heart are key factors in preventing the surge of O2·- production upon reoxygenation.

Silky dogwood (Swida amomum) seed dispersal by freshwater turtles.

PREMISE: Effective seed dispersal is essential to the success of plant species. Swida amomum (silky dogwood) has a seed-dispersal syndrome characteristic of autumn-ripening shrubs with fleshy fruits; attached fruits are ingested and defecated by birds, while fallen fruits are consumed by ground-foraging birds and mammals. METHODS: We documented that fallen fruits of this shrub were consumed by two aquatic turtle species (eastern painted turtle [Chrysemys picta] and red-eared slider [Trachemys scripta]) and that their seeds were defecated. We compared germination success (percentage of seeds germinated) of defecated seeds, seeds collected from a pond surface, and seeds removed from shrubs. RESULTS: While four seed species were identified in fecal samples, seeds of S. amomum were the most frequent (93%) among samples and the most numerous (106 seeds) in any sample. Average proportion of fecal seeds germinated (85.99%) exceeded that of seeds from the pond surface (82.76%) and from shrubs (60.24%), albeit the difference in germination success was insignificant. When analyzed using fecal samples from painted turtles only, the difference in germination success between fecal seeds and those collected from pond or shrub became significant. CONCLUSIONS: Our findings represent the first report of S. amomum seeds being dispersed by turtle gut passage and suggest aquatic turtles could be an important part of a secondary seed dispersal process influencing woody plant community composition in temperate wetland ecosystems.

Divergent incubation temperature effects on thermal sensitivity of hatchling performance in two different latitudinal populations of an invasive turtle.

The incubation temperature for embryonic development affects several aspects of hatchling performance, but its impact on the thermal sensitivity of performance attributes remains poorly investigated. In the present study, Trachemys scripta elegans hatchlings from two different latitudinal populations were collected to assess the effects of different incubation temperatures on the locomotor (swimming speed) and physiological (heart rate) performances, and the thermal sensitivity of these two attributes. The incubation temperature significantly affected the examined physiological traits. Hatchling turtles produced at low incubation temperature exhibited relatively higher cold tolerance (lower body temperatures at which the animals lose the ability to escape from the lethal conditions), and reduced heart rate and swimming speed. Furthermore, the effect of incubation temperature on the thermal sensitivity of swimming speed differed between the low- and high-latitude populations. At relatively high incubation temperatures, the high-latitude hatchling turtles exhibited reduced thermal sensitivities of swimming speed than those of the low-latitude ones. Reduced thermal sensitivity of locomotor performance together with high cold tolerance, exhibited by the high-latitude hatchling turtles potentially reflected local adaptation to relatively colder and more thermally-variable environments.

Can colored object enrichment reduce the escape behavior of captive freshwater turtles?

The effect of environmental enrichment on the behavior and welfare in captivity of reptiles and of freshwater turtles in particular, which are popular aquarium and pet species, is very little studied compared to other taxa. We carried out a small scale case-study on the effect of colored object enrichment, with and without fish scent, on the behavior of a group of 15 cooters (Pseudemys sp.) and sliders (Trachemys scripta ssp.) on display at a public aquarium. The new enrichment aimed to reduce the escape behavior (interaction with transparent boundaries) and increase exploration and random swimming. We used simultaneous recording of behavior at whole group level and for focal individually-marked turtles. The escape behavior decreased on days with new enrichment before feeding at whole group level and for the focal turtles overall, in spite of the relatively low interest in the colored objects. Fish-scented objects attracted significantly more interest. Random swimming, enrichment focus, aggression and submission increased significantly, and basking decreased significantly at whole group level before feeding, with smaller differences after feeding. There were large differences between individual turtles with respect to activity budgets and changes in behavior on days with new enrichment, with both increases and decreases seen in escape behavior, aggression, and levels of activity. Our outcomes suggested that introducing new colored objects with food scent may be beneficial for reducing escape behavior in captive freshwater turtles. However, careful monitoring of effects at individual level and much larger scale investigations, including postenrichment periods, are needed.

Dmrt1 induces the male pathway in a turtle species with temperature-dependent sex determination.

The molecular mechanism underlying temperature-dependent sex determination (TSD) has been a long-standing mystery; in particular, the thermosensitive genetic triggers for gonadal sex differentiation are largely unknown. Here, we have characterized a conserved DM domain gene, Dmrt1, in the red-eared slider turtle Trachemys scripta (T. scripta), which exhibits TSD. We found that Dmrt1 has a temperature-dependent, sexually dimorphic expression pattern, preceding gonadal sex differentiation, and is capable of responding rapidly to temperature shifts and aromatase inhibitor treatment. Most importantly, loss- and gain-of-function analyses provide solid evidence that Dmrt1 is both necessary and sufficient to initiate male development in T. scripta Furthermore, the DNA methylation dynamics of the Dmrt1 promoter are tightly correlated with temperature and could mediate the impact of temperature on sex determination. Collectively, our findings demonstrate that Dmrt1 is a candidate master male sex-determining gene in this TSD species, consistent with the idea that DM domain genes are conserved during the evolution of sex determination mechanisms.

Genetic variation and admixture of red-eared sliders (Trachemys scripta elegans) in the USA.

The most ubiquitous, abundant, and invasive turtle on Earth, Trachemys scripta elegans (TSE, "red-eared slider"), is one of four taxa in a clade that is native to the USA and adjacent Mexico (three subspecies of Trachemys scripta plus Trachemys gaigeae). The present range-wide study of this clade is based on 173 known-locality mtDNA sequences combined with ddRAD libraries for 43 samples emphasizing the western part of the range of TSE, its contact with that of T. gaigeae, and anthropogenic hybrids between TSE and T. s. scripta. The data presented here are the first to sample the TSE × T. s. scripta intergrade zone or TSE × T. s. scripta crosses from introduced turtles. In the western part of its range (New Mexico and Texas), most samples of TSE from the Pecos River have mtDNA haplotypes matching T. gaigeae. Structure analysis of SNPs from the ddRAD show evidence of genetic admixture between T. gaigeae and TSE in all included samples from the Rio Grande and Pecos River. These populations also exhibit T. gaigeae-like head stripes, i.e., a postorbital marking that does not reach the eye. The genetic and morphological data are thereby reconciled, as both suggest that these TSE are intergrades. We recommend that these populations continue to be considered TSE, despite the admixture with T. gaigeae. In the Eastern United States, some samples of the morphologically intermediate subspecies T. s. troostii are not genetically distinct from TSE and some samples share morphological characters and genetic affinities with T. s. scripta. Based on these observations we conclude that the taxon T. s. troostii represents intergrades between TSE and T. s. scripta and should not be considered a valid taxon. Near the already established part of the intergrade zone between TSE and T. s. scripta, TSE mtDNA haplotypes have naturally introgressed into typical-looking samples of T. s. scripta in Georgia. Hybrids between introduced TSE and T. s. scripta are also confirmed deeper within the natural range of T. s. scripta in South Carolina and Virginia. Given the examples of feral hybrids deep within its range shown here and elsewhere, the threat of genetic pollution of T. s. scripta by feral TSE is established.

Induction of foxo3a protects turtle neurons against oxidative stress.

The detrimental effects of oxidative stress caused by the accumulation of Reactive Oxygen Species (ROS) factor into aging, senescence and several neurodegenerative diseases. Mammalian models are extremely susceptible to the stresses that follow the restoration of oxygen after anoxia; however some organisms including the freshwater turtle Trachemys scripta can withstand extended anoxia and reoxygenation without apparent pathology. The ability of the turtle to withstand these conditions is thought to be linked to the upregulation of protective mechanisms such as heat shock proteins (HSP) as well as the suppression of ROS formation and the upregulation of antioxidant defenses. One such antioxidant mechanism is the transcription factor Forkhead box O3a (FOXO3a), that has been shown to be activated in several animal models during oxidative stress. In this study, we utilized both the transfection of a plasmid carrying foxo3a and the pharmacological manipulation of foxo3a using the green tea extract Epigallocatechin-3-gallate (EGCG) to investigate the protective role of FOXO3a in the turtle brain. Our studies found that transcript levels of foxo3a were upregulated significantly during reoxygenation with greater increases during chemical oxidative stress. Induction of foxo3a by direct transfection significantly decreased cell death during chemical oxidative stress. Cells treated with EGCG also showed increased foxo3a expression and decreased cell death in the presence of H2O2. These results agree with results seen in other animal models and suggest that EGCG (through the upregulation of foxo3a) may be a therapeutic target against oxidative stress damage that warrants further investigation.

Pharmacokinetics of injectable marbofloxacin after intravenous and intramuscular administration in red-eared sliders (Trachemys scripta elegans).

Fluoroquinolone antibacterial drugs are currently used in reptilian medicine because of their broad spectrum of activity including the most frequent pathogens of these species. The disposition kinetics of marbofloxacin (MBX) at a single dose of 2 mg/kg were determined in healthy red-eared sliders after intravenous (IV) and intramuscular (IM) administration. The influence of renal portal system on the bioavailability of the drug was investigated by using forelimb and hindlimb as IM injection sites. Apparent volume of distribution at steady-state (Vss ) and systemic clearance (Cl) of marbofloxacin after IV administration were estimated to be 48.21 ± 5.42 ml/kg and 23.38 ± 2.90 ml/hr·kg, respectively. The absolute bioavailabilities after IM route were 45.96% (forelimb) and 52.09% (hindlimb). The lack of statistically significant differences in most of the pharmacokinetic parameters after the two IM injection sites suggests a negligible influence of renal portal system in clinical use of MBX, although the Cmax after IMfore administration is advantageous, having into account the concentration-dependent action of this antibiotic. The absence of visible adverse reactions in the animals and the advantageous pharmacokinetic properties suggest the possibility of its safe and effective clinical use in red-eared sliders.

TEMPORAL STABILITY OF IN VITRO VENOUS BLOOD GAS, pH, AND LACTATE VALUES OF COWNOSE RAYS (RHINOPTERA BONASUS) AND RED-EARED SLIDER TURTLES (PSEUDEMYS SCRIPTA ELEGANS).

This study assessed the in vitro temporal changes that occur in blood pH and lactate concentrations for an elasmobranch species and a chelonian species, as well as blood gases (partial pressures of carbon dioxide [pCO2] and oxygen [pO2]) for a chelonian species, with a portable clinical point-of-care analyzer. Blood samples were collected from 10 cownose rays (Rhinoptera bonasus) and 10 red-eared sliders (Pseudemys scripta elegans), stored on ice, and serially analyzed at six time points up to 90 min postcollection. Results indicate that analysis should be conducted as soon as possible after blood collection for these species, with immediate analysis being preferred. However, if analysis must be delayed, syringes may be capped, placed on ice, and analyzed at a later time. Analysis within 90 min provided clinically acceptable results for pH and lactate in both species and for pCO2 in red-eared sliders, whereas substantial artifactual increases of pO2 were seen in red-eared sliders.

Auditory brainstem responses in the red-eared slider Trachemys scripta elegans (Testudoformes: Emydidae) reveal sexually dimorphic hearing sensitivity.

Hearing sensitivity is of general interest from the perspective of understanding the functionality and evolution of vertebrate auditory systems. Sexual dimorphism of auditory systems has been reported in several species of vertebrates, but little is known about this phenomenon in turtles. Some morphological characteristics, such as middle ear and tympanic membrane that influence the hearing sensitivity of animals can result in hearing sexual dimorphism. To examine whether sexual dimorphism in hearing sensitivity occurs in turtles and to compare hearing characteristics with respect to the shape of the tympanic membrane, we measured the hearing sensitivity and tympanum diameter in both sexes of Trachemys scripta elegans. The results showed that, with the exception of 0.9 kHz, auditory brainstem response thresholds were significantly lower in females than in males for frequencies in the 0.2-1.1 kHz range, indicating that the hearing of females shows greater sensitivity. No significant differences were detected in the tympanum diameter of both sexes. These results showed that sexually dimorphic hearing sensitivity has evolved in turtles; however, this difference does not appear to be related to differences in the size of the tympanic membrane. The possible origin and function of the sexual differences in auditory characteristic are discussed.

Tissue-dependent variation of hydrogen sulfide homeostasis in anoxic freshwater turtles.

Hydrogen sulfide (H2S) controls numerous physiological responses. To understand its proposed role in metabolic suppression, we measured free H2S and bound sulfane sulfur (BSS) in tissues of the freshwater turtle Trachemys scriptaelegans, a species undergoing strong metabolic suppression when cold and anoxic. In warm normoxic turtles, free H2S was higher in red blood cells (RBCs) and kidney (∼9-10 µmol l-1) than in brain, liver and lung (∼1-2 µmol l-1). These values overall aligned with the tissue H2S-generating enzymatic activity. BSS levels were similar in all tissues (∼0.5 µmol l-1) but ∼100-fold higher in RBCs, which have a high thiol content, suggesting that RBCs function as a circulating H2S reservoir. Cold acclimation caused significant changes in free and bound H2S in liver, brain and RBCs, but anoxia had no further effect, except in the brain. These results show tissue-dependent sulfide signaling with a potential role in brain metabolic suppression during anoxia in turtles.

INTRAVENOUS LIPID EMULSION TREATMENT REDUCES SYMPTOMS OF BREVETOXICOSIS IN TURTLES (TRACHEMYS SCRIPTA).

Harmful algal blooms (HABs) occur when excess nutrients allow dinoflagellates to reproduce in large numbers. Marine animals are affected by blooms when algal toxins are ingested or inhaled. In the Gulf of Mexico, near annual blooms of Karenia brevis release a suite of compounds (brevetoxins) that cause sea turtle morbidity and mortality. The primary treatment at rehabilitation facilities for brevetoxin-exposed sea turtles is supportive care, and it has been difficult to design alternative treatment strategies without an understanding of the effects of brevetoxins in turtles in vivo. Previous studies using the freshwater turtle as a model species showed that brevetoxin-3 impacts the nervous and muscular systems, and is detoxified and eliminated primarily through the liver, bile, and feces. In this study, freshwater turtles (Trachemys scripta) were exposed to brevetoxin (PbTx-3) intratracheally at doses causing clear systemic effects, and treatment strategies aimed at reducing the postexposure neurological and muscular deficits were tested. Brevetoxin-exposed T. scripta displayed the same behaviors as animals admitted to rehabilitation centers for toxin exposure, ranging from muscle twitching and incoordination to paralysis and unresponsiveness. Two treatment regimes were tested: cholestyramine, a bile acid sequestrant; and an intravenous lipid emulsion treatment (Intralipidt) that provides an expanded circulating lipid volume. Cholestyramine was administered orally 1 hr and 6 hr post PbTx-3 exposure, but this regime failed to increase toxin clearance. Animals treated with Intralipid (100 mg/kg) 30 min after PbTx-3 exposure had greatly reduced symptoms of brevetoxicosis within the first 2 hr compared with animals that did not receive the treatment, and appeared fully recovered within 24 hr compared with toxin-exposed control animals that did not receive Intralipid. The results strongly suggest that Intralipid treatment for lipophilic toxins such as PbTx-3 has the potential to reduce morbidity and mortality in HAB-exposed sea turtles.

Evidence of embryonic regulation of maternally derived yolk corticosterone.

In recent years, the potential for maternal stress effects to adaptively alter offspring phenotype has received considerable attention. This research has identified offspring traits that are labile in response to maternal stress; however, an understanding of the mechanisms underlying these effects is lagging and is crucial to appreciating the significance of this maternal effect. In the present study, we sought to better understand maternal stress effects by examining the potential for embryonic regulation of corticosterone exposure, determining the phenotypic consequences of elevated corticosterone during development, and characterizing the levels of maternally transferred corticosterone in unmanipulated eggs using Trachemys scripta By dosing eggs with tritiated corticosterone and tracking the steroid throughout development, we found that most corticosterone is metabolized, and less than 1% of the corticosterone dose reaches the embryo as free corticosterone. We also found that exogenous dosing of corticosterone, in concentrations sufficient to overwhelm embryonic metabolism, reduces embryonic survival and negatively impacts hatchling traits important to fitness. Our results demonstrate that concentrations of maternal corticosterone in the yolks of unmanipulated eggs are low and are significantly lower than the doses of corticosterone required to elicit phenotypic effects in hatchlings. Taken together, these results provide evidence that both the embryo and the female may minimize corticosterone accumulation in the embryo to avoid reductions in embryonic survival and negative impacts on offspring phenotype and fitness.

Experimentally Induced Selenosis in Yellow-Bellied Slider Turtles ( Trachemys scripta scripta).

Selenosis, or selenium toxicosis, occurs in wildlife and livestock, usually because of excessive intake of selenium via selenium-containing plants. Although it is known that wild slider turtles can accumulate large amounts of selenium, little is known about how selenium exposure may affect these reptiles. In this study, the authors report histopathologic changes in yellow-bellied sliders ( Trachemys scripta scripta) caused by experimental exposure to selenomethionine. Microscopic changes in kidney and claw tissue were most significant and resembled those reported in birds. Turtles in the selenium treatment groups had acute tubular degeneration and regeneration in the kidney, with hyaline droplets in the high-dose animals, and changes in the claws ranging from epidermal hyperplasia with disorganization and intercellular edema to ulceration, and accumulation of seroheterophilic exudate between the epidermis and cornified layer. Although selenium burdens in this study are comparable with values found in wild slider turtles, more data are needed to determine if similar histopathologic abnormalities arise in wild animals exposed to high levels of selenium.

The high resolution melting analysis (HRM) as a molecular tool for monitoring parasites of the wildlife.

In an interconnected world, the international pet trade on wild animals is becoming increasingly important. As a consequence, non-native parasite species are introduced, which affect the health of wildlife and contribute to the loss of biodiversity. Because the investigation of parasite diversity within vulnerable host species implies the molecular identification of large samples of parasite eggs, the sequencing of DNA barcodes is time-consuming and costly. Thereby, the objectives of our study were to apply the high resolution melting (HRM) approach for species determination from pools of parasite eggs. Molecular assays were validated on flatworm parasites (polystomes) infecting the Mediterranean pond turtle Mauremys leprosa and the invasive red-eared slider Trachemys scripta elegans in French natural environments. HRM analysis results indicated that double or multiple parasitic infections could be detected from wild animal populations. They also showed that the cycle of parasite eggs production was not regular over time and may depend on several factors, among which the ecological niche and the target species. Thereby, monitoring parasites from wild endangered animals implies periodic parasitological surveys to avoid false negative diagnostics, based solely on eggs production.

Effects of topical insulin on second-intention wound healing in the red-eared slider turtle (Trachemys scripta elegans) - a controlled study.

BACKGROUND: Compared with mammals, wound healing in reptiles is characterized by reduced wound contraction and longer healing times. The aim of this study is to describe the clinical and histopathological effects of topical insulin on second-intention healing of experimentally induced wounds in skin without dermal bony plates of Trachemys scripta elegans exposed to daily variations in ambient temperature and in an aquatic environment. Forty-four healthy adult females were assigned to two groups: Group 1 (n = 24) was used to assess clinical features such as wound contraction; Group 2 (n = 20) was used for histological evaluation and morphometric analysis. Topical porcine insulin (5 IU/ml diluted in glycerol) was applied daily 1 week. For each control time (2, 7, 14, 21 and 28 days post-wounding), re-epithelisation and wound remodelling were evaluated histologically and the number of main inflammatory cells (heterophils, macrophages, lymphocytes and fibroblasts) was scored. RESULTS: Mean wound contraction was higher in the insulin-treated group at each time point and differences were significant at day 28 (P < 0.0001). Histologically, these clinical findings were associated with better re-epithelisation, inflammatory response, collagen synthesis and remodelling of the wounds. Morphometrically, insulin-treated wounds had significantly higher mean counts of heterophils (day 7), macrophages (days 2, 7 and 14) and fibroblasts (days 14 and 21), whereas lymphocyte counts were significantly lower at day 21. These results demonstrate that topical insulin modifies the inflammatory response of turtle skin up-regulating inflammatory cells at early stages and promoting wound healing. CONCLUSIONS: Topical insulin is a potentially useful therapy in skin wounds of Trachemys scripta and should be evaluated in non-experimental wounds of turtles and other reptiles.

Effects of deslorelin acetate on plasma testosterone concentrations in captive yellow-bellied sliders (Trachemys scripta sp.).

In Europe, the yellow-bellied slider (Trachemys scripta sp.) is a non-native species in competition with native freshwater turtles. Research on contraception could be useful to control the captive population. Identifying a method of contraception in chelonians would potentially help to control aggression in other chelonian species. The objective of the current study was to evaluate the effects of a single 4.7-mg deslorelin acetate implant on plasma testosterone concentrations in yellow-bellied sliders (Trachemys scripta sp.). Eleven adult male yellow-bellied sliders were used for the study. Males from the treatment group (n = 6) received a 4.7-mg deslorelin acetate implant, whereas males from the control group (n = 5) did not receive any treatment. All individuals were housed under the same environmental conditions. Testosterone plasma concentrations of the control group and the treatment group were measured at six time points (T0-T6) between April and September. No difference between the control group and the deslorelin treatment group was observed at T0, T2, T3, T4, T5 or T6. However, mean plasma testosterone concentration was significantly higher in the treatment group than in the control group at T1. This suggests that treatment with a 4.7-mg deslorelin acetate implant has a transient stimulatory effect on the anterior pituitary in yellowbellied sliders without a negative feedback on testosterone production. Further studies with a higher dosage of deslorelin acetate are needed to draw conclusions on its contraceptive effect.

Continuous arterial PO2 profiles in unrestrained, undisturbed aquatic turtles during routine behaviors.

Mammals and birds maintain high arterial partial pressure of oxygen (PO2 ) values in order to preserve near-complete hemoglobin (Hb) oxygen (O2) saturation. In diving mammals and birds, arterial O2 follows a primarily monotonic decline and then recovers quickly after dives. In laboratory studies of submerged freshwater turtles, arterial O2 depletion typically follows a similar pattern. However, in these studies, turtles were disturbed, frequently tethered to external equipment and confined either to small tanks or breathing holes. Aquatic turtles can alter cardiac shunting patterns, which will affect arterial PO2  values. Consequently, little is known about arterial O2 regulation and use in undisturbed turtles. We conducted the first study to continuously measure arterial PO2  using implanted microelectrodes and a backpack logger in undisturbed red-eared sliders during routine activities. Arterial PO2  profiles during submergences varied dramatically, with no consistent patterns. Arterial PO2  was also lower than previously reported during all activities, with values rarely above 50 mmHg (85% Hb saturation). There was no difference in mean PO2  between five different activities: submerged resting, swimming, basking, resting at the surface and when a person was present. These results suggest significant cardiac shunting occurs during routine activities as well as submergences. However, the lack of relationship between PO2  and any activity suggests that cardiac shunts are not regulated to maintain high arterial PO2  values. These data support the idea that cardiac shunting is the passive by-product of regulation of vascular resistances by the autonomic nervous system.