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The effects of dietary vitamin C on growth, liver vitamin C and serum cortisol in stressed and unstressed juvenile soft-shelled turtles (Pelodiscus sinensis).

We studied the effect of dietary vitamin C on growth, liver vitamin C and serum cortisol levels in stressed and unstressed juvenile soft-shelled turtles. Turtles were fed with vitamin C supplementation at dosages of 0, 250, 500, 2500, 5,000 or 10,000 mg/kg diet for 4 weeks. Vitamin C supplementation exerted significant effects on specific growth rate and liver vitamin C concentrations. The specific growth rate peaked in the group fed at 500 mg/kg diet, while liver vitamin C levels increased with increasing dietary vitamin C levels. Serum cortisol concentrations did not differ between groups of turtles fed diets supplemented with vitamin C in the range of 0-10,000 mg/kg. After acid stress, hepatic vitamin C levels were unaffected, while serum cortisol in the control group was significantly elevated (P<0.01). The other five groups of turtles did not show significant changes in serum cortisol compared with pre-stress levels.

Animals↗

Ganglion cell types of the turtle retina that project to the optic tectum: Intracellular HRP injections of retrogradely, rhodamine-marked cell bodies.

The turtle retina has been shown to have a variety of different morphological ganglion cell types as well as distinct physiological ganglion cell types. The major projection of the retina to the brain in nonmammalian vertebrates is to the optic tectum. In this study, we address the question of which retinal ganglion cell types project to the optic tectum in the turtle. Fluorescent rhodamine-labeled microspheres were used to trace the retinal ganglion cell projection to the superficial layers of the optic tectum. The fluorescent ganglion cell somata, retrogradely marked by transport from the contralateral optic tectum, were impaled with micropipettes containing rhodamine-horseradish peroxidase solution and this dye was iontophoresed into the cells under visual control. Most of the morphological ganglion cell types described in Golgi studies (Kolb, 1982; Kolb et al., 1988) were stained. Thus, the small cell types G1, G2, G3, G5, G6, and G7; the medium-sized types G10, G11, G12, G13, and G14; and the large-sized types G15, G16, G19, G20, and G21 project to the optic tectum in the turtle. We have added a new type, G2a, which proves to have some differences from the original G2 in branching pattern. We were unable to stain the small type G4, the medium-sized types G8 and G9, and the large cell types G17 and G18: this suggests that they might not project to the superficial layers of the dorsolateral optic tectum, at least, in the turtle.

Animals↗

Spectral properties of short-wavelength (blue) cones in the turtle retina.

Long- and medium-wavelength cones in the turtle retina participate in complex neural interactions. They are coupled via excitatory pathways to other cones and receive negative feedback inputs from luminosity-type horizontal cells. Little information has been collected on the S- (short-wavelength or blue) cones because they are scarce in the turtle retina and of smaller dimensions compared to the other cone types. In this paper, flash sensitivity action spectra and photoresponses of seven turtle S-cones were measured in the dark-adapted state and during chromatic background illuminations. The desensitizing action of monochromatic background lights was not uniform across the visible spectrum. A red background was most effective in desensitizing the S-cones to long-wavelength stimuli while a blue background light produced its strongest action on the photoresponses elicited by short-wavelength stimuli. The effects of chromatic adaptation on the S-cone action spectrum and on the kinetics of the small-amplitude photoresponses suggested that the S-cones in the turtle retina were involved in complex neural interactions. These included excitatory inputs probably originating in neighboring L-cones and inhibitory long-wavelength inputs probably mediated by L-type horizontal cells.

Absorption↗

Immunocytochemical and histochemical localization of nitric oxide synthase in the turtle retina.

Recent interest in nitric oxide and its relationship to cGMP has produced many attempts to anatomically localize the enzyme synthesizing nitric oxide, nitric oxide synthase. In the retina, numerous previous studies have used the NADPH-diaphorase enzyme activity of nitric oxide synthase as a histochemical method to localize nitric oxide synthase. However, all NADPH-diaphorase activity is not necessarily nitric oxide synthase, because several enzymes have similar biochemical activity. Additionally, various histochemical methods have been used to demonstrate NADPH-diaphorase activity, which makes comparisons between studies difficult. The purpose of this study was twofold. First, we wanted to examine the histochemical labeling of NADPH-diaphorase in the turtle retina to allow comparisons to previous studies. Second, we wanted to compare the histochemical localization of NADPH-diaphorase activity to the immunocytochemical localization of nitric oxide synthase in the turtle retina. Our histochemical localization of NADPH-diaphorase activity and our localization of nitric oxide synthase-like immunoreactivity in the turtle retina both produced similar results. Both the histochemistry and immunocytochemistry consistently labeled photoreceptor inner segments, at least three amacrine cell types, and processes in the inner plexiform layer. In optimized double-labeled preparations, all cells with NADPH-diaphorase activity were also positive for nitric oxide synthase-like immunoreactivity, although some somata in the ganglion cell layer only had nitric oxide synthase-like immunoreactivity. The immunocytochemical localization of nitric oxide synthase in photoreceptors, amacrine cells, and putative ganglion cells indicates that nitric oxide may function at several levels of visual processing in the turtle retina.

Animals↗

Antioxidant systems and anoxia tolerance in a freshwater turtle Trachemys scripta elegans.

The effects of anoxic submergence (20 h at 5 degrees C) and subsequent 24 h aerobic recovery on the antioxidant systems of six organs were examined in freshwater turtles, Trachemys scripta elegans. Both xanthine oxidase and xanthine dehydrogenase were detected in turtle tissues with xanthine oxidase composing 36-75% of the total activity. Turtle organs displayed high constitutive activities of catalase (CAT), superoxide dismutase (SOD), and alkyl hydroperoxide reductase (AHR). Measurements of lipid peroxidation damage products (conjugated dienes, lipid hydroperoxides, thiobarbituric acid reactive substances) showed minimal changes during anoxia or recovery suggesting that natural anoxic-aerobic transitions occur without the free radical damage that is seen during ischemia-reperfusion in mammals. Anoxia exposure led to selected decreases in enzyme activities in organs, consistent with a reduced potential for oxidative damage during anoxia: SOD decreased in liver by 30%, CAT decreased in heart by 31%, CAT and total glutathione peroxidase (GPOX) decreased in kidney (by 68 and 41%), and CAT and SOD decreased in brain (by 80 and 15%). AHR, however, increased 2 and 3.5 fold during anoxia in heart and kidney respectively. Most anoxia-induced changes were reversed during aerobic recovery although brain enzyme activities remained suppressed. Some specific changes occurred during the recovery period: SOD increased from controls in heart by 45%, AHR increased to 200 and 168% of control values in red and white muscle respectively, and total GPOX decreased from controls in heart and white muscle by 75 and 77% respectively. The results show that biochemical adaptation for natural anoxia tolerance in turtles includes well-developed antioxidant defenses that minimize or prevent damage by reactive oxygen species during the reoxygenation of organs after anoxic submergence.

Acclimatization↗

Multiple paternity in loggerhead turtle (Caretta caretta) nests on Melbourne Beach, Florida: a microsatellite analysis.

Many aspects of sea turtle biology are difficult to measure in these enigmatic migratory species, and this lack of knowledge continues to hamper conservation efforts. The first study of paternity in a sea turtle species used allozyme analysis to suggest multiple paternity in loggerhead turtle (Caretta caretta) clutches in Australia. Subsequent studies indicated that the frequency of multiple paternity varies from species to species and perhaps location to location. This study examined fine-scale population structure and paternal contribution to loggerhead clutches on Melbourne Beach, FL, USA using microsatellite markers. Mothers and offspring from 70 nests collected at two locations were analysed using two to four polymorphic microsatellite loci. Fine-scale population differentiation was not evident between the sampled locations, separated by 8 km. Multiple paternity was common in loggerhead nests on Melbourne Beach; 22 of 70 clutches had more than one father, and six had more than two fathers. This is the first time that more than two fathers have been detected for offspring in individual sea turtle nests. Paternal genotypes could not be assigned with confidence in clutches with more than two fathers, leaving the question of male philopatry unanswered. Given the high incidence of multiple paternity, we conclude that males are not a limiting resource for this central Florida nesting aggregate.

Animals↗

Polyandry in a marine turtle: females make the best of a bad job.

The female perspective on reproductive strategies remains one of the most active areas of debate in biology. Even though a single mating is often sufficient to satisfy the fertilization needs of most females and the act of further mating incurs costs, multiple paternity within broods or clutches is a common observation in nature. Direct or indirect advantage to females is the most popular explanation. However, the ubiquity of this explanation is being challenged by an increasing number of cases for which benefits are not evident. For the first time, we test possible fitness correlates of multiple paternity in a marine turtle, an organism that has long attracted attention in this area of research. Contrary to the wide-spread assumption that multiple mating by female marine turtles confers fitness benefits, none were apparent. In this study, the environment played a far stronger role in determining the success of clutches than whether paternity had been single or multiple. A more likely explanation for observations of multiply sired clutches in marine turtles is that these are successful outcomes of male coercion, where females have conceded to superfluous matings as a compromise. Thus, multiple matings by female marine turtles may be a form of damage control as females attempt to make the best of a bad job in response to male harassment.

Animals↗

Identification and initial characterization of serum growth hormone binding protein in the turtle Chrysemys dorbigni.

Proteins that bind growth hormone (GHBP) have been identified in the blood of many mammalian and avian species, but not in reptilian species. We carried out binding studies with the serum of turtles using chromatographic techniques as well as the dextran-charcoal separation method. As in other species, we found at least two different GHBPs: one with high MW and low affinity and the other with lower MW and higher affinity. The high affinity GHBP was partially purified using gel filtration and affinity chromatography, reaching a degree of purification of 11,000 times (0.17 nmol/g of serum protein in the serum vs 1900 nmol/g protein in the purified material). When the high affinity GHBP was characterized, it was found to have a dissociation constant (Kd: 2.6 +/- 0.7 nM) similar to those described for mouse or rat, but lower than those for chicken, rabbit or man. The binding capacity (Bmax) was 120 +/- 43 fmoles/mg of protein, which can be also expressed as 1.08 +/- 0.38 pmol/ml of serum. A preliminary MW estimation of 50-60 kDa was obtained for turtle higher affinity GHBP. The specificity of this high affinity GHBP is somatogenic, since bovine GH competes as well as human GH for 125I-hGH bound to binding protein, while ovine PRL competes only partially and with low affinity. Unrelated hormones, as insulin and glucagon, can not displace the 125I-hGH bound to turtle GHBP. A very important seasonal variation in turtle GHBP activity was observed: maximum binding was found in November (springtime), followed by a continuous decline over March and May.

Animals↗

Multiple data sets, high homoplasy, and the phylogeny of softshell turtles (Testudines: Trionychidae).

We present a phylogenetic hypothesis and novel, rank-free classification for all extant species of softshell turtles (Testudines:Trionychidae). Our data set included DNA sequence data from two mitochondrial protein-coding genes and a approximately 1-kb nuclear intron for 23 of 26 recognized species, and 59 previously published morphological characters for a complimentary set of 24 species. The combined data set provided complete taxonomic coverage for this globally distributed clade of turtles, with incomplete data for a few taxa. Although our taxonomic sampling is complete, most of the modern taxa are representatives of old and very divergent lineages. Thus, due to biological realities, our sampling consists of one or a few representatives of several ancient lineages across a relatively deep phylogenetic tree. Our analyses of the combined data set converge on a set of well-supported relationships, which is in accord with many aspects of traditional softshell systematics including the monophyly of the Cyclanorbinae and Trionychinae. However, our results conflict with other aspects of current taxonomy and indicate that most of the currently recognized tribes are not monophyletic. We use this strong estimate of the phylogeny of softshell turtles for two purposes: (1) as the basis for a novel rank-free classification, and (2) to retrospectively examine strategies for analyzing highly homoplasious mtDNA data in deep phylogenetic problems where increased taxon sampling is not an option. Weeded and weighted parsimony, and model-based techniques, generally improved the phylogenetic performance of highly homoplasious mtDNA sequences, but no single strategy completely mitigated the problems of associated with these highly homoplasious data. Many deep nodes in the softshell turtle phylogeny were confidently recovered only after the addition of largely nonhomoplasious data from the nuclear intron.

Animals↗

Ecological correlates and evolutionary divergence in the skull of turtles: a geometric morphometric assessment.

Resource use and phylogeny are often correlated with morphological variation. Moreover, because biological shapes are often complex and evolve depending on several internal constraints, they must be assessed using integrative methods. We analyzed the morphological variation of the turtle skull in the context of an adaptive radiation. Our focus are turtles of the superfamily Testudinoidea, which are remarkably diverse, both in number of species and in ecology. In this study, we depict morphological variation in the turtle skull in three dimensions with respect to diet, phylogeny, and habitat using modern geometric morphometrics. Our study revealed that morphological specialization was related to both diet and habitat. Morphological variation is decomposed in regard of both resource use (habitat and diet) and phylogeny. Feeding mode depending on environment was suggested as a key factor determining morphological evolution and diversification of turtle skulls. Diet (especially durophagy) leads to parallel morphologies in different clades. Phylogeny seemed to constrain only localized features of the skull and remained of minor influence, because overall morphotypes, closely correlated with ecological factors, occurred in both clades. In conclusion, the adaptive radiation of the Testudinoidea is revealed to demonstrate a clear relationship between the skull shape and life style.

Adaptation, Physiological↗

Response to dietary dilution in an omnivorous freshwater turtle: implications for ontogenetic dietary shifts.

Several species of freshwater turtles in the family Emydidae undergo an ontogenetic dietary shift; as juvenile turtles mature, they change from a primarily carnivorous to a primarily herbivorous diet. It has been hypothesized that this shift results from an unfavorable ratio of gut capacity to metabolic rate that prevents small reptiles from processing adequate volumes of plant material to meet their energetic demands. Effects of dietary dilution on intake were evaluated in two size classes of red-eared sliders (Trachemys scripta elegans) to test whether small reptiles have a lower capacity to compensate for low-quality diets through increased intake than do larger conspecifics. Artificial diets with an inert diluent were offered to two size classes of turtles, and mass-specific intakes of dry matter, energy, and nitrogen were calculated. Both small (28.7+/-4.9 g body mass, mean mass+/-SD) and large (1,230+/-94 g body mass) turtles compensated for dietary dilution and maintained constant energy and nitrogen intakes on diets with lower energy content than common aquatic plants. Thus, body size did not affect the ability to respond to nutritional dilution, which suggests that processing limitations imposed by small body size do not constrain juveniles from adopting an herbivorous diet.

Animals↗

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.

Adaptation, Physiological↗

Survival and physiological responses of hatchling blanding's turtles (Emydoidea blandingii) to submergence in normoxic and hypoxic water under simulated winter conditions.

Overwintering habits of hatchling Blanding's turtles (Emydoidea blandingii) are unknown. To determine whether these turtles are able to survive winter in aquatic habitats, we submerged hatchlings in normoxic (155 mmHg Po2) and hypoxic (6 mmHg Po2) water at 4 degrees C, recording survival times and measuring changes in key physiological variables. For comparison, we simultaneously studied hatchling softshell (Apalone spinifera) and snapping (Chelydra serpentina) turtles, which are known to overwinter in aquatic habitats. In normoxic water, C. serpentina and A. spinifera survived to the termination of the experiment (76 and 77 d, respectively). Approximately one-third of the E. blandingii died during 75 d of normoxic submergence, but the cause of mortality was unclear. In hypoxic water, average survival times were 6 d for A. spinifera, 13 d for E. blandingii, and 19 d for C. serpentina. Mortality during hypoxic submergence was probably caused by metabolic acidosis, which resulted from accumulated lactate. Unlike the case with adult turtles, our hatchlings did not increase plasma calcium and magnesium, nor did they sequester lactate within the shell. Our results suggest that hatchling E. blandingii are not particularly well suited to hibernation in hypoxic aquatic habitats.

Acclimatization↗

Ontogenetic scaling of bite force in lizards and turtles.

Because selection on juvenile life-history stages is likely strong, disproportionately high levels of performance (e.g., sprint speed, endurance, etc.) might be expected. Whereas this phenomenon has been demonstrated with respect to locomotor performance, data for feeding are scarce. Here, we investigate the relationships among body dimensions, head dimensions, and bite force during growth in lizards and turtles. We also investigate whether ontogenetic changes in bite performance are related to changes in diet. Our analyses show that, for turtles, head dimensions generally increase with negative allometry. For lizards, heads scale as expected for geometrically growing systems. Bite force generally increased isometrically with carapace length in turtles but showed significant positive allometry relative to body dimensions in lizards. However, both lizards and turtles display positive allometric scaling of bite force relative to some measures of head size throughout ontogeny, suggesting (1) strong selection for increased relative bite performance with increasing head size and (2) intrinsic changes in the geometry and/or mass of the jaw adductors during growth. Whereas our data generally do not provide strong evidence of compensation for lower absolute levels of performance, they do show strong links among morphology, bite force, and diet during growth.

Animals↗

Cold tolerance in hatchling painted turtles (Chrysemys picta): supercooling or tolerance for freezing?

We studied tolerance for cold in hatchling painted turtles (Chrysemys picta) from Lake Metigoshe, Bottineau County, North Dakota, to determine whether neonates in populations near the northern limit of distribution rely on a tolerance for freezing or on a capacity for supercooling to survive their first winter of life. We placed hatchlings individually into artificial hibernacula constructed in jars of damp, loamy sand and then cooled the jars to approximately -0.45 degrees C, which was below the equilibrium freezing point for water held by the sand but above that for body fluids of the neonatal turtles. A piece of ice next was placed on the surface of the sand in each jar to induce freezing of the soil water. After the soil water had frozen to an equilibrium, the temperature in the jars was lowered by 1 degrees C/d to minima averaging -2.5 degrees C, -4.5 degrees C, -6.5 degrees C, and -10.5 degrees C in different treatments. These temperatures were maintained for varying periods, so that animals in each treatment were exposed to temperatures below the equilibrium freezing point for their body fluids for a total of 11 d. Thirty of 32 hatchlings survived exposure to -2.5 degrees C; 24 of 32 survived at -4.5 degrees C; 14 of 32 withstood -6.5 degrees C; and 7 of 32 tolerated -10.5 degrees C. Freezing exotherms were detected in temperature profiles for turtles that succumbed but not in those for hatchlings that survived. Thus, the ability of hatchlings to withstand subzero temperatures for extended periods apparently requires that they avoid freezing. Although other workers contend that tolerance for freezing is the key to survival over winter by hatchling painted turtles from the region of Lake Metigoshe, our findings indicate that neonates rely primarily on their ability to remain unfrozen and supercooled.

Animals↗

Testing dispersal hypotheses in foraging green sea turtles (Chelonia mydas) of Brazil.

Testing theories of dispersal is challenging in highly migratory species. In sea turtles, population size, geographic distance, natal homing, and ocean currents are hypothesized to affect dispersal. Little is known, however, about these mechanisms in sea turtles foraging along the South American coast. Green sea turtles feeding at Ubatuba (UB, n = 114) and Almofala (AF, n = 117), Brazil, were sequenced at the mitochondrial DNA (mtDNA) control region (486 bp) and genotyped at 7 microsatellite loci to test dispersal hypotheses. Fifteen mtDNA haplotypes were revealed, including a previously undescribed sequence, and the average observed heterozygosity (H(o)) was 76.4%. Overall short-term temporal differences were not detected, and differentiation was less pronounced in microsatellite than in mtDNA analyses. Mitochondrial results reveal significant differentiation between the Brazilian feeding grounds and most other Atlantic groups, whereas microsatellites uncover similarities to some of the geographically closest populations. Ubatuba and Almofala are mixed stocks, drawn primarily from Ascension, with lesser contributions from Surinam/Aves and Trindade. Costa Rica is also a significant source of individuals feeding at AF. The results are consistent with a model of juvenile natal homing impacted by other factors. Effective protection of turtles foraging along the extensive Brazilian coast may enhance breeding populations thousands of kilometers away.

Animal Migration↗

First application of the SINE (short interspersed repetitive element) method to infer phylogenetic relationships in reptiles: an example from the turtle superfamily Testudinoidea.

Although turtles (order Testudines) constitute one of the major reptile groups, their phylogenetic relationships remain largely unresolved. Hence, we attempted to elucidate their phylogeny using the SINE (short interspersed repetitive element) method, in which the sharing of a SINE at orthologous loci is indicative of synapomorphy. First, a detailed characterization of the tortoise polIII/SINE was conducted using 10 species from eight families of hidden-necked turtles (suborder Cryptodira). Our analysis of 382 SINE sequences newly isolated in the present study revealed two subgroups, namely Cry I and Cry II, which were distinguishable according to diagnostic nucleotides in the 3' region. Furthermore, four (IA-ID) and five (IIA-IIE) different SINE types were identified within Cry I and Cry II subgroups, respectively, based on features of insertions/deletions located in the middle of the SINE sequences. The relative frequency of occurrence of the subgroups and the types of SINEs in this family were highly variable among different lineages of turtles, suggesting active differential retroposition in each lineage. Further application of the SINE method using the most retrotranspositionally active types, namely IB and IC, challenged the established phylogenetic relationships of Bataguridae and its related families. The data for 11 orthologous loci demonstrated a close relationship between Bataguridae and Testudinidae, as well as the presence of the three clades within Bataguridae. Although the SINE method has been used to infer the phylogenies of a number of vertebrate groups, it has never been applied to reptiles. The present study represents the first application of this method to a phylogenetic analysis of this class of vertebrates, and it provides detailed information on the SINE subgroups and types. This information may be applied to the phylogenetic resolution of relevant turtle lineages.

Amino Acid Sequence↗

Reproductive deficits in male freshwater turtle Chrysemys picta from Cape Cod, Massachusetts.

Contaminated groundwater plumes have formed on the Massachusetts Military Reservation (MMR), a Superfund site on Cape Cod, Massachusetts, as a result of chemical waste disposal. The plumes are of concern to the local people who rely on groundwater as a drinking water source. We used the freshwater turtle as a sentinel species to monitor the reproductive effects of exposure and, by inference, the potential for impact on human health. Our observations of male Chrysemys picta field-trapped from Moody Pond (an impacted site) and Washburn Pond (a reference site) on Cape Cod extended and supported prior observations of reproductive deficits. Morphometric comparison of precloacal length (PCL), which is a sexually dimorphic trait in the turtle, showed that Moody Pond males had a significantly longer PCL than Washburn Pond males. Moody Pond turtles showed reduced testicular weight, which was associated with significantly smaller seminiferous tubule diameter. Epididymal sperm counts were also markedly reduced in Moody Pond animals compared to Washburn Pond animals. Testicular histology and gonial proliferation, as determined by PCNA, were similar in both male populations, while the Moody Pond males had significantly higher germ cell apoptosis than the animals in Washburn Pond. These results suggest that a low-level mixture of xenobiotic contaminants impairs the reproductive functions of turtles exposed to the impacted site but not to the reference site environment.

Animals↗