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Turtles and tortoises (chelonians) have been integral components of global ecosystems for about 220 million years and have played important roles in human culture for at least 400,000 years. The chelonian shell is a remarkable evolutionary adaptation, facilitating success in terrestrial, freshwater and marine ecosystems. Today, more than half of the 360 living species and 482 total taxa (species and subspecies combined) are threatened with extinction. This places chelonians among the groups with the highest extinction risk of any sizeable vertebrate group. Turtle populations are declining rapidly due to habitat loss, consumption by humans for food and traditional medicines and collection for the international pet trade. Many taxa could become extinct in this century. Here, we examine survival threats to turtles and tortoises and discuss the interventions that will be needed to prevent widespread extinction in this group in coming decades., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Life histories evolve in response to constraints on the time available for growth and development. Nesting date and its plasticity in response to spring temperature may therefore be important components of fitness in oviparous ectotherms near their northern range limit, as reproducing early provides more time for embryos to complete development before winter. We used data collected over several decades to compare air temperature and nest date plasticity in populations of painted turtles and snapping turtles from a relatively warm environment (southeastern Michigan) near the southern extent of the last glacial maximum to a relatively cool environment (central Ontario) near the northern extent of post-glacial recolonization. For painted turtles, population-level differences in reaction norm elevation for two phenological traits were consistent with adaptation to time constraints, but no differences in reaction norm slopes were observed. For snapping turtle populations, the difference in reaction norm elevation for a single phenological trait was in the opposite direction of what was expected under adaptation to time constraints, and no difference in reaction norm slope was observed. Finally, among-individual variation in individual plasticity for nesting date was detected only in the northern population of snapping turtles, suggesting that reaction norms are less canalized in this northern population. Overall, we observed evidence of phenological adaptation, and possibly maladaptation, to time constraints in long-lived reptiles. Where present, (mal)adaptation occurred by virtue of differences in reaction norm elevation, not reaction norm slope. Glacial history, generation time, and genetic constraint may all play an important role in the evolution of phenological timing and its plasticity in long-lived reptiles., (© 2016 by the Ecological Society of America.)

Portions of the reproductive tract of the male (Trachemys scripta) turtle were examined by immunocytochemistry for evidence of the capacity to produce and respond to estrogen hormones (via the expression of P450 aromatase and estrogen receptors). Aromatase was detected in both the Sertoli and Leydig cells of the testis and was expressed at different levels during the spermatogenic cycle, being highest in the quiescent testis and lowest during germ cell meiosis. ERalpha was found in the Leydig cells surrounding the seminiferous tubules as well as in the epithelial cells of the excurrent canals (rete testis, efferent ductule, and epididymis). ERbeta immunoreactivity was found in both the spermatogonia and Sertoli cells in the testis, and in the epithelial cells of excurrent canals.

Measuring standard metabolic rate (SMR) and specific dynamic action (SDA) has yielded insight into patterns of energy expenditure in snakes, but less emphasis has been placed on identifying metabolic variation and associated energy cost of circadian rhythms. To estimate SMR, SDA, and identify metabolic variation associated with circadian cycles in nocturnally active African house snakes (Lamprophis fuliginosus), we measured oxygen consumption rates (VO2) at frequent intervals before and during digestion of meals equaling 10%, 20% and 30% of their body mass. Circadian rhythms in metabolism were perceptible in the VO2 data during fasting and after the initial stages of digestion. We estimated SMR of L. fuliginosus (mean mass=16.7+/-0.3 g) to be 0.68+/-0.02 (+/-SEM) mL O2/h at 25 degrees C. Twenty-four hours after eating, VO2 peaked at 3.2-5.3 times SMR. During digestion of meals equaling 10-30% of their body mass, the volume of oxygen consumed ranged from 109 to 119 mL O2 for SMR, whereas extra oxygen consumed for digestion and assimilation ranged from 68 to 256 mL O2 (equivalent to 14.5-17.0% of ingested energy). The oxygen consumed due to the rise in metabolism during the active phase of the daily cycle ranged from 55 to 66 mL O2 during digestion. Peak VO2, digestive scope, and SDA increased with increasing meal size. Comparisons of our estimates to estimates derived from methods used in previous investigations resulted in wide variance of metabolic variables (up to 39%), likely due to the influence of circadian rhythms and activity on the selection of baseline metabolism. We suggest frequent VO2 measurements over multiple days, coupled with mathematical methods that reduce the influence of undesired sources of VO2 variation (e.g., activity, circadian cycles) are needed to reliably assess SMR and SDA in animals exhibiting strong circadian cycles.

Previous laboratory studies indicate that coal combustion wastes (a mixture composed of fly ash and other lower volume wastes such as bottom ash; hereafter collectively referred to as ash) adversely affect the health of benthic fish (Erimyzon sucetta; lake chubsucker), but fish in these studies were provided with ample uncontaminated food resources. Because aquatic disposal of ash can also adversely affect food resources for benthic fish, we hypothesized that changes in resources might exacerbate the effects of ash on fish observed in laboratory studies. We exposed juvenile E. sucetta in outdoor microcosms to water, sediment, and benthic resources from an ash-contaminated site or a reference site for 45 days and compared our findings to previous laboratory studies. Benthic invertebrate biomass was nearly three times greater in controls compared to ash microcosms. Total organic content of control sediment (41%) was also greater than in ash sediments (17%), suggesting that additional benthic resources may have also been limited in ash microcosms. Benthic invertebrates isolated from the ash microcosms had trace element concentrations (As, Cd, Co, Cr, Cs, Se, Sr, and V) up to 18 times higher than in weathered ash used in laboratory studies. The concentrations of trace elements accumulated by fish reflected the high dietary concentrations encountered in the ash microcosms and were associated with reduced growth (final mass = 0.07 g) and survival (25%) compared to controls (0.37 g and 67%, respectively). Accumulation of trace elements, as well as reductions in growth and survival, were more pronounced than in previous laboratory studies, suggesting that resource conditions may be important in mediating ash toxicity. Taken together, our studies suggest that ash discharge into aquatic systems is a more serious threat to the health of benthic fish than previously predicted based upon laboratory toxicity tests.

Coal fly ash and effluent from coal ash settling basins negatively affects metabolism and reproduction in a variety of organisms, including a number of fish species. Some species, most notably the eastern mosquitofish (Gambusia holbrooki), are known to maintain viable populations in areas contaminated by coal ash. While eastern mosquitofish are present in these systems, their degree of tolerance to coal ash has not been investigated using sublethal metrics of exposure. It is possible that eastern mosquitofish persist in habitats affected by coal ash, but experience significant costs such as changes in metabolism and fecundity. Thus, we investigated the effects of coal ash on standard metabolic rate and reproduction of eastern mosquitofish inhabiting a coal-ash contaminated settling basin. Standard metabolic rates of mosquitofish from the ash contaminated site and a reference site (mean O2 consumption = 0.286 mL/g x h +/- 0.007 and 0.291 +/- 0.008 mL/g x h, respectively) were not significantly different. Despite elevated contaminant concentrations in ash basin females (selenium, arsenic, copper, and cadmium) and their offspring (selenium), brood sizes and offspring viability did not differ between clutches collected from ash basin and reference site females. Our data provide further evidence of the high degree of tolerance of eastern mosquitofish to exposure to aquatic coal ash disposal generated by power plants. However, the basis for such tolerance to ash remains unclear. Further investigations are required to determine whether such tolerance is a result of species-specific characteristics or population characteristics due to local adaptation.

Estimating energy costs by respirometry is fundamental to many studies of the ecology, behavior and evolution of reptiles. However, traditional respirometry procedures seldom incorporate objective techniques for removal of outliers from estimates of metabolic parameters. We demonstrate how computer-automated respirometry equipment, which records many respiratory measurements over short intervals, can be coupled with mathematical procedures to produce robust estimates of pre- and post-prandial metabolism in banded water snakes (Nerodia fasciata fasciata). Standard metabolic rate of N. f. fasciata was estimated to be 1.21 ml O2/h (mass = 30.21 +/- 0.74 g) at 25 degrees C. After ingestion of a fish equaling 20% of their body mass, snakes exhibited a fivefold increase in metabolic rate with peak O2 consumption rate (VO2) reaching 6.5 ml O2/h. Total cost of digestion was 5.44 kJ, equivalent to approximately 21% of the energy in the meal. Repeated measurements of metabolism in the same individuals revealed that our methods yielded similar results, even when individuals exhibited different patterns of VO2 variation between respiratory trials. Our results underscore the importance of obtaining many VO2 measurements, coupled with objective removal of outlier values from estimates of metabolic rate, especially when metabolic values are to be interpreted in a comparative context.

Although the amount of energy that males and females invest in reproduction is an integral component of theories explaining the evolution of particular mating strategies, few studies have actually determined the amount of energy that each sex allocates to reproduction. We compared how energy is expended by male and female Anolis carolinensis lizards during both the breeding and postbreeding seasons. We used laboratory respirometry to determine resting metabolic rates (RMRs) of inactive, freshly captured lizards and the doubly labeled water technique to determine field metabolic rates (FMRs) of free-ranging lizards. Both RMRs and FMRs were influenced by body mass but not by sex. Season did not influence FMRs; however, RMRs of both sexes increased approximately 40% from the breeding to the postbreeding season. The seasonal increase in RMRs was attributed to a postreproductive increase in feeding rate and specific dynamic action. We used RMRs, FMRs, and thermal profiles of lizards to calculate energy budgets for breeding and postbreeding seasons. Energy budgets partitioned daily field energy (DFE; calculated from FMRs) into daily activity energy (DAE) and daily resting energy (DRE; calculated from RMRs). Energy expended for reproduction was estimated as DAE during the breeding season plus egg production (for females). Despite males having 40% greater body mass, females expended 46% more energy for reproduction than did males (906 and 619 J/d, respectively). Total metabolizable energy (TME=DFE+egg production for females) expended during the breeding season was similar for males and females (1,280 and 1,365 J/d, respectively). Although TME of females decreased 44% from the breeding to the postbreeding season (1,365 vs. 766 J/d), TME of males was similar during both seasons (1,280 vs. 1,245 J/d). There were both seasonal and sexual differences in DRE and DAE. Compared with most lizards from semiarid/desert habitats, A. carolinensis in a temperate habitat expends more total energy during the breeding season, allocates more energy to eggs, and appears to have more total energy available for reproduction.

During metamorphic climax, anuran larvae must rely on stored energy because changes in oral and digestive morphology prevent foraging and efficient assimilation. Thus, the time required to store adequate energy for metamorphic climax may set a lower limit on age at which it can occur. Therefore, the amount and type of energy used during metamorphic climax must be determined. To quantify the energetic costs of metamorphic climax in Bufo terrestris, oxygen consumption during climax was measured. Wet mass, dry mass, and lipid mass for a group of individuals at the initiation of climax (forelimb emergence, FL) and for another group at the end of climax (complete tail resorption, TR) were also measured to determine whether lipids were used to fuel metamorphic climax. The total amount of energy used, maintenance costs, and development costs during metamorphic climax varied considerably among individuals. Variation in energy metabolism during climax was not related to differences in energy metabolism during larval development or body mass at initiation of climax. TR individuals were significantly lighter in terms of wet mass and had less body water than FL individuals. However, the two groups did not differ in dry mass or lipid mass. Therefore, lipid catabolism is not a major source of energy during metamorphic climax in B. terrestris. As a result, decreases in age at metamorphosis may not be constrained by the need to store energy in the form of lipids.

For 38 of the past 50 years, Painted Turtles were studied on the University of Michigan's E.S. George Reserve in southeastern Michigan. We compared age specific body sizes, reproductive traits and survival of Painted Turtles ranging from 9 to 61 years of age to test contrasting predictions of the Relative Reproductive Rate and Senescence Hypotheses of aging. Indeterminate growth (i.e. continued body growth of adults) was important in increasing reproductive output of older turtles; however, growth rate of the oldest age-group was reduced compared to that of younger adults. Although clutch size and among year reproductive frequency did not increase with age, within year reproductive frequency (production of second clutches), egg size, and hatchling size did. Nest predation rates and the proportion of surviving nests that produced hatchlings were similar among age groups, and embryo mortality in nests was not related to age. Survivorship of males was less than that of females, and survivorship of the oldest group of females was not statistically different from that of a younger group of females. No decline in reproductive output or survivorship was detected in the oldest females as predicted by the Senescence Hypothesis. Thus, the majority of data on reproductive traits and survivorship support the Relative Reproductive Rate Hypothesis. We also compared Painted Turtles to Blanding's Turtles, another species studied on the E.S. George Reserve. That Painted Turtles exhibit indeterminate growth whereas Blanding's Turtles do not, appears to be a primary mechanism for some differences between species in the relationships between reproductive traits and age. An important mechanism for increasing reproductive output in both species was increased reproductive frequency in older females. Painted Turtles also increased offspring quality (egg and hatchling size) with age, whereas Blanding's Turtles did not. Compared to younger individuals, there was no reduction in survivorship in the oldest Painted Turtles and survivorship increased in the oldest Blanding's Turtles.

Numerous field surveys suggest that coal combustion wastes (ash) adversely affect fish populations, but few controlled laboratory studies have evaluated the responses of individual fish to ash exposure. Available information suggests that sublethal effects of ash, including decreased growth rates and reduced fecundity, may be important manifestations of ash toxicity. Here, we hypothesized that ash may also alter swimming performance, which could ultimately have important implications for the autecology of affected species. To test this hypothesis, we measured sprint speed and critical swimming speed (U(crit)) of juvenile lake chubsuckers (Erimyzon sucetta) exposed to ash in the laboratory. Fish exposed to ash for 90-100 days accumulated significant concentrations of As, Se, Sr, and V; exhibited severe fin erosion; and had reduced sprint speed and U(crit). Compared to controls, sprint speed of ash-exposed fish was reduced by 30% at 5 cm and the percent reduction was further reduced to 104% at 20 cm. Critical swimming speed was approximately 50% lower in fish exposed to ash compared to controls. Additionally, the typical positive relationship between standard length and U(crit) was absent in fish exposed to ash. Because reductions in swimming performance could not be attributed to pollutant-induced differences in body condition or fin morphology, we hypothesize that physiological disruptions (e.g., increased energy demands, decreased oxygen uptake or transport) are responsible for the observed effects and warrant further attention.

The germ cell development in the slider turtle (Trachemys scripta) testis was investigated by viewing the histology of the seminiferous epithelium in plastic sections with a light microscope. Germ cell morphologies in the slider turtle testis were similar to the morphologies of other vertebrate germ cell types. However, the slider turtle seminiferous epithelium contained germ cells that progress through spermatogenesis in a temporal rather than a spatial pattern, resulting in a single spermatogenic event that climaxed with one massive sperm release in November. Mature sperm then are stored within the epididymis until breeding commences in the following spring. The germ cell development strategy in the slider turtle is different from that of other amniotes and is more reminiscent of the developmental strategy found in the anamniotic testis. This temporal progression of germ cells through spermatogenesis within a tubular testis represents a transitional model that may be evolutionarily significant., (Copyright 2003 Wiley-Liss, Inc.)

Biodosimetry, the estimation of received doses by determining the frequency of radiation-induced chromosome aberrations, is widely applied in humans acutely exposed as a result of accidents or for clinical purposes, but biodosimetric techniques have not been utilized in organisms chronically exposed to radionuclides in contaminated environments. The application of biodosimetry to environmental exposure scenarios could greatly improve the accuracy, and reduce the uncertainties, of ecological risk assessments and biomonitoring studies, because no assumptions are required regarding external exposure rates and the movement of organisms into and out of contaminated areas. Furthermore, unlike residue analyses of environmental media, environmental biodosimetry provides a genetically relevant biomarker of cumulative lifetime exposure. Symmetrical chromosome translocations can impact reproductive success, and could therefore prove to be ecologically relevant as well. We describe our experience in studying aberrations in the yellow-bellied slider turtle as an example of environmental biodosimetry., (Copyright 2002 Elsevier Science Ltd.)

We provide an overview of research related to environmental effects of disposal of coal combustion residues (CCR) in sites in the United States. Our focus is on aspects of CCR that have the potential to negatively influence aquatic organisms and the health of aquatic ecosystems. We identify major issues of concern, as well as areas in need of further investigation. Intentional or accidental release of CCR into aquatic systems has generally been associated with deleterious environmental effects. A large number of metals and trace elements are present in CCR, some of which are rapidly accumulated to high concentrations by aquatic organisms. Moreover, a variety of biological responses have been observed in organisms following exposure to and accumulation of CCR-related contaminants. In some vertebrates and invertebrates, CCR exposure has led to numerous histopathological, behavioral, and physiological (reproductive, energetic, and endocrinological) effects. Fish kills and extirpation of some fish species have been associated with CCR release, as have indirect effects on survival and growth of aquatic animals mediated by changes in resource abundance or quality. Recovery of CCR-impacted sites can be extremely slow due to continued cycling of contaminants within the system, even in sites that only received CCR effluents for short periods of time. The literature synthesis reveals important considerations for future investigations of CCR-impacted sites. Many studies have examined biological responses to CCR with respect to Se concentrations and accumulation because of teratogenic and reproductively toxic effects known to be associated with this element. However, the complex mixture of metals and trace elements characteristic of CCR suggests that biological assessments of many CCR-contaminated habitats should examine a variety of inorganic compounds in sediments, water, and tissues before causation can be linked to individual CCR components. Most evaluations of effects of CCR in aquatic environments have focused on lentic systems and the populations of animals occupying them. Much less is known about CCR effects in lotic systems, in which the contaminants may be transported downstream, diluted or concentrated in downstream areas, and accumulated by more transient species. Although some research has examined accumulation and effects of contaminants on terrestrial and avian species that visit CCR-impacted aquatic sites, more extensive research is also needed in this area. Effects in terrestrial or semiaquatic species range from accumulation and maternal transfer of elements to complete recruitment failure, suggesting that CCR effects need to be examined both within and outside of the aquatic habitats into which CCR is released. Requiring special attention are waterfowl and amphibians that use CCR-contaminated sites during specific seasons or life stages and are highly dependent on aquatic habitat quality during those periods. Whether accidentally discharged into natural aquatic systems or present in impoundments that attract wildlife, CCR appears to present significant risks to aquatic and semiaquatic organisms. Effects may be as subtle as changes in physiology or as drastic as extirpation of entire populations. When examined as a whole, research on responses of aquatic organisms to CCR suggests that reducing the use of disposal methods that include an aquatic slurry phase may alleviate some environmental risks associated with the waste products.

Study organisms in chronic toxicological bioassays are often provided with excessive resources to remove food limitations as a confounding experimental variable. Under more ecologically realistic situations, resources are often less abundant and such restrictions may alter the responses of organisms to environmental contaminants. Here, we investigated the interaction between resource level and sediment toxicity in the lake chubsucker, Erimyzon sucetta. For 78 days we fed fish one of three ration levels (1X, 2X, 4X; uncontaminated food) that was grazed directly from either clean sand or coal ash-contaminated sediments. Despite provision of uncontaminated food, fish exposed to the contaminated sediments accumulated significant whole body concentrations of As, Se, Sr, and V. Food ration affected the pattern of Se accumulation, with lowest concentrations accumulated by fish supplied with the lowest rations (1X). Paradoxically, fish in the 1X-ash treatment were most adversely effected by ash-exposure, despite having Se burdens much lower than fish in the 2X- and 4X-ash treatments. Fish in the 1X-ash treatment exhibited higher mortality, lower proportional growth, and increased incidence of fin erosion compared to fish provided with higher rations. Such results may, in part, be explained by the apparent inability of fish with reduced rations to maintain positive energy balance, as evidenced by their higher standard metabolic rates compared to control fish fed similar rations. Our results underscore the importance of considering resource quantity and nutritional factors in chronic bioassays in order to draw more ecologically realistic conclusions about contaminant effects.

Little currently is known about the accumulation or effects of contaminants on reptiles. To date, most studies examining reptile exposure to trace elements report tissue burdens of field-captured animals, but seldom provide insight into the dose, duration, or mode of exposure involved. For two years, we fed juvenile banded water snakes (Nerodia fasciata) prey items collected from a coal ash-contaminated site that contained elevated levels of As, Cd, Cu, Se, Sr, and V. With the exception of Cu, snakes accumulated significant concentrations of elements, usually in a dose-dependent manner. Accumulation varied significantly among liver, kidney, and gonads, and in most cases between sexes. Selenium accumulation was most notable, greatly exceeding established toxicity thresholds for other vertebrates. Despite the high concentrations of pollutants accumulated, snakes exposed to the contaminated diet survived through the study and exhibited normal food consumption, growth, condition factor, overwinter survival and mass loss, metabolic rate, and gonadosomatic index. The results of this study confirm that diet can be a significant route of exposure to trace elements in snakes and indicate that further studies on snakes are warranted to better understand their responses to contaminants.

The storage and behavior of sperm collected from the epididymis of two emyiid turtles were examined. In Chrysemys picta, the weight of the epididymis does not change significantly throughout the year as does the testis. However, in this species, as well as in Trachemys scripta, the epididymis contains sperm throughout the entire year. Sperm from both species have a relatively low motility and velocity of movement. In C. picta, equally low motilities are observed both in the autumn, shortly after spermiation, and in spring. Motility could be augmented by the addition of isobutyl methyl xanthine (IBMX, 0.5 mmol l(-1)). Epididymal sperm remained viable in excess of 40 days in vitro when stored in F-10 buffer, during which time motility and swimming velocity could be augmented with IBMX. The longevity and low motility of turtle sperm facilitates its storage by either males or females, and creates conditions that promote the wide dissemination, over time, of gametes produced in a narrow time window., (Copyright 2002 Wiley-Liss, Inc.)

Habitat contamination can alter numerous biological processes in individual organisms. Examining multiple individual-level responses in an integrative fashion is necessary to understand how individual health or fitness reflects environmental contamination. Here we provide an example of such an integrated perspective based upon recent studies of an amphibian (the bullfrog, Rana catesbeiana ) that experiences several, disparate changes when larval development occurs in a trace element-contaminated habitat. First, we present an overview of studies focused on specific responses of individuals collected from, or transplanted into, a habitat contaminated by coal combustion residues (CCR). These studies have reported morphological, behavioral, and physiological modifications to individuals chronically interacting with sediments in the CCR-contaminated site. Morphological abnormalities in the oral and tail regions in contaminant-exposed individuals influenced other properties such as grazing, growth, and swimming performance. Behavioral changes in swimming activities and responses to stimuli appear to influence predation risk in the contaminant-exposed population. Significant changes in bioenergetics in the contaminated habitat, evident as abnormally high energetic expenditures for survival (maintenance) costs, may ultimately influence production pathways (growth, energy storage) in individuals. We then present a conceptual model to examine how interactions among the affected systems (morphological, behavioral, physiological) may ultimately bring about more severe effects than would be predicted if the responses were considered in isolation. A complex interplay among simultaneously occurring biological changes emerges in which multiple, sublethal effects ultimately can translate into reductions in larval or juvenile survival, and thus reduced recruitment of juveniles into the population. In systems where individuals are exposed to low concentrations of contaminants for long periods of time, research focused on one or few sublethal responses could substantially underestimate overall effects on individuals. We suggest that investigators adopt a more integrated perspective on contaminant-induced biological changes so that studies of individual-based effects can be better integrated into analyses of mechanisms of population change.

Whereas ecological assessments of contaminants are concerned with populations and higher levels of organization, most mechanistic work in toxicology is directed at effects on individuals and their parts. We propose that studies based on individuals can be useful in ecological analysis of polluted systems when based on the concepts of resource allocation-based life history analysis. At the heart of the resource allocation approach is the concept of operative environments of individuals (i.e., environmental factors influencing birth, death, or migration). Contaminants can have strong influences on operative environments, modifying resource allocation strategies that reflect changes in energy assimilation and demands. By examining contaminant-induced responses of individuals from the perspective of changing operative environments, individual-based changes and population dynamics can be addressed in an ecologically rigorous manner.

Recent studies of several vertebrates and an invertebrate have shown elevated standard metabolic rate (SMR) following chronic exposure to a mixture of trace elements in a contaminated habitat. In this study, we examined whether another invertebrate, a crayfish (Procambarus acutus), also experienced elevated SMR in response to the same contaminants. We compared SMR of individuals inhabiting the contaminated site with SMR of individuals from uncontaminated reference sites. We also examined SMR of individuals collected from the reference areas and exposed in the laboratory for 50 days to sediment and food derived from the contaminated site. Individuals collected from the contaminated site had elevated SMR compared to individuals collected from the unpolluted areas (25.1 vs. 19.2 J g(-1) day(-1)). Individuals exposed to contaminated sediment and food in the laboratory experienced elevations in SMR compared to controls after 27 days of exposure (35.2 vs. 29.4 J g(-1) day(-1)), but after 50 days of exposure, metabolic rate no longer differed between treatments. Growth of contaminant-exposed individuals was lower than growth of reference animals throughout the laboratory study. Elevated SMR associated with contaminant exposure may reflect energy-demanding mechanisms required to combat deleterious effects of contaminants. Our results support the prediction that increases in energy expenditure in the contaminated habitat would negatively influence production processes, such as growth. Results from this study in conjunction with observations from other species suggest that increased SMR is a common response among several taxa to the mixture of contaminants in the study site.

Coal combustion wastes are enriched in a number of potentially toxic compounds and may pose risks to biota exposed to the wastes. Slider turtles (Trachemys scripta) are common inhabitants of coal ash settling basins in South Carolina, USA, where they feed on contaminated prey items and accumulate high levels of potentially toxic compounds in their tissues. Furthermore, female sliders sometimes nest in contaminated spill piles and thus may expose embryos to contaminated soils. We examined two potential pathways by which female T. scripta may influence the survivorship and quality of their offspring in a contaminated habitat: (1) nesting in contaminated soil and (2) maternal transfer of pollutants. Eggs were collected from turtles captured in coal ash-polluted or unpolluted sites; individual clutches were incubated in both ash-contaminated and uncontaminated soil in outdoor, artificial nests. Incubation in contaminated soil was associated with reduced embryo survivorship. Adult females from the polluted site accumulated high levels of As, Cd, Cr, and Se in their tissues, yet Se was the only element transferred maternally to hatchlings at relatively high levels. Hatchlings from polluted-site females exhibited reduced O2 consumption rates compared to hatchlings from reference sites. Relatively high levels of Se transferred to hatchlings by females at the ash-polluted site might contribute to the observed differences in hatchling physiology.

For 35 of the past 47 years, Blanding's turtles were studied on the University of Michigan's E.S. George Reserve in southeastern Michigan. Blanding's turtle is one of the longest-lived emydid turtles with individuals reaching ages greater than 75 years. We compared body sizes, reproductive traits and survival of Young, Middle, and Oldest age groups of Blanding's turtles to test predictions from two contrasting hypotheses of aging. The relative reproductive rate hypothesis predicts traits that increase the reproductive output or survival rates of older compared to younger individuals, whereas the senescence hypothesis predicts a reduction in reproductive output or survival in older versus younger individuals. Body size did not increase with age among groups; therefore, indeterminate growth was not a mechanism for increased reproductive output of the oldest individuals. Survivorship, reproductive frequency and size-adjusted mean clutch size were all higher in the Oldest age group compared to the younger age groups. Nest predation rate was highest in the Young age group compared to either group of older turtles. In nests that survived predation, the proportion of nests that failed entirely due to developmental problems was lowest in the Young, intermediate in the Middle, and highest in the Oldest age group. Successful nests produced similar numbers of hatchlings and similar sized hatchlings in all three age groups. Traits such as egg and offspring size, and offspring produced per nest did not support either the relative reproductive rate or the senescence hypothesis of aging. Increased embryo mortality in nests of older females compared to younger turtles supports predictions from the senescence hypothesis. Three traits; increased clutch size, reproductive frequency, and survivorship of individuals in the Oldest age group compared to younger turtles support the relative reproductive rate hypothesis for evolution of longevity.

Using a whole-chromosome FISH painting probe we previously developed for chromosome 1 of the yellow-bellied slider turtle (Trachemys scripta), we investigated the dose-rate effect for radiation-induced symmetrical translocations in T. scripta fibroblasts and lymphocytes. The dose rate below which no reduction in effect per unit dose is observed with further dose protraction was approximately 23 cGy h(-1). We estimated the whole-genome spontaneous background level of complete, apparently simple symmetrical translocations in T. scripta lymphocytes to be approximately 1.20 x 10(-3)/cell projected from aberrations occurring in chromosome 1. Similar spontaneous background levels reported for humans are some 6- to 25-fold higher, ranging from about 6 x 10(-3) to 3.4 x 10(-2) per cell. This relatively low background level for turtles would be a significant advantage for resolution of effects at low doses and dose rates. We also chronically irradiated turtles over a range of doses from 0-8 Gy delivered at approximately 5.5 cGy h(-1) and constructed a lymphocyte dose-response curve for complete, apparently simple symmetrical translocations suitable for use with animals chronically exposed to radiation in contaminated environments. The best-fitting calibration curve (not constrained through the zero dose estimate) was of the form Y(as) = c + aD + bD(2), where Y(as) was the number of apparently simple symmetrical translocations per cell, D was the dose (Gy), a = (0.0058 +/- 0.0009), b = (-0.00033 +/- 0.00011), and c = (0.0015 +/- 0.0013). With additional whole-chromosome probes to improve sensitivity, environmental biodosimetry using stable chromosome translocations could provide a practical and genetically relevant measurement end point for ecological risk assessments and biomonitoring programs.

Fluorescence in situ hybridization of a whole chromosome 1-specific probe from the yellow-bellied slider turtle (Trachemys scripta) to cells from four other species of turtle ranging from a desert tortoise to a loggerhead sea turtle resulted in specific and exclusive hybridization to chromosome 1 in all five species. Previous observations of conservation in the giemsa banding pattern and chromosome morphology and number among turtles are thus extended to the DNA sequence level, revealing a cytogenetic stability of chromosome 1 in these turtles during the past 66-144 million years. This contrasts with the situation for various hominoid species where, in many instances, extensive chromosomal rearrangements have been reported in one third of that time period. Our probe, which was prepared by microdissecting whole chromosomes from embryonic T. scripta fibroblasts and amplifying using DOP-PCR, is the first report of a whole-chromosome FISH probe for any reptile.

Compared with birds, mammals, fish, and even amphibians, very little is known about the effects of contaminants on reptiles. Recent evidence that many reptile populations may be declining has stimulated demand for toxicological studies of reptiles as well as development of nondestructive sampling techniques useful for assessing and monitoring contaminant exposure. The current study experimentally evaluated the utility of shed skins, tail clips, and blood samples as nondestructive indices of trace element exposure in banded water snakes, Nerodia fasciata. For 13.5 months, snakes were either fed fish from a coal ash-contaminated site or uncontaminated food from a reference site. Snakes fed contaminated prey accumulated As, Cd, Se, Sr, and V in various organs (i.e. liver, kidney, and/or gonads). Moreover, non-parametric discriminant function analysis revealed that snakes could be placed in two groups that reliably reflected their experimental diet based upon Se, Sr, and As concentrations in tail clips, blood, and/or shed skins. We suggest that nondestructive sampling techniques, particularly analyses of blood and tail clips, may be easily applied in evaluations of contaminant exposure in the field and laboratory and may prevent excessive destructive sampling of potentially threatened reptile species.

Lake chubsuckers (Erimyzon sucetta) were exposed to coal ash-polluted sediments under conservative experimental conditions (filtered artificial soft water and abundant uncontaminated food). After 4 months of exposure, fish grazing the polluted sediments had significantly elevated body burdens of Se, Sr, and V. Selenium levels were particularly elevated, reaching mean whole body concentrations of 5.6 microg/g dry mass by the end of experimental manipulations. Twenty-five percent of fish exposed to pollutants died during the study. All surviving fish exposed to ash exhibited substantial decreases in growth and severe fin erosion. Total nonpolar lipids were two times higher in fish from the control treatment, but percent lipid did not differ between treatments. Because fish were presented with the same amount of food during the study, it appears fish exposed to ash utilized more energy for daily activities and/or were less efficient at converting available energy to tissues for growth and storage. The results were particularly interesting because we were unable to detect differences in standard metabolic rate (SMR) of fish between treatments. Increased energy expenditures not detectable in estimates of maintenance based on SMR, such as costs of digestion or activity, may have contributed to decreased energetic efficiency. Our findings corroborate previous studies which have documented the toxicity of ash-derived pollutants in fish.

The viability and motility of spermatozoa harvested from the epididymides of turtles were estimated to elucidate properties that might enable them to be stored over long periods of time. Spermatozoa from the painted turtle, Chrysemys picta, were analysed and compared with spermatozoa from two other turtles, Trachemys scripta and Sternotherus odoratus using the Cellsoft analysis system for videotaped images. Spermatozoa from C. picta and T. scripta, suspended in F-10 medium, showed low motility (3-6% motile) and motion velocities, whereas the motility of spermatozoa from S. odoratus was higher (40% motile). Spermatozoa from C. picta and S. odoratus, but not T. scripta, had higher motilities and motion velocities when incubated at 2 degrees C before analyses. C. picta spermatozoa were unresponsive to calcium concentrations ranging from 10(-8) to 10(-1) mol l(-1), potassium concentrations ranging from 0. 1 to 10 mmol l(-1), and to pH values in the range 5.9-8.4. Spermatozoa from C. picta were sensitive to hypo-osmotic media, and showed reduced motility at 25% of normal osmolarity and no motility at 10% of normal osmolarity. Distorted cells and missing flagellae were noted at 50% of normal osmolarity. C. picta spermatozoa were viable up to 40 days after harvest when incubated at 4 degrees C; during this time, both motility and motion velocity were increased in response to 0.5 mmol 3-isobutyl-1-methylxanthine l(-1). Spermatozoa from turtles have osmotic properties and resistance to changing chemical environments similar to spermatozoa from other vertebrates that have internal fertilization, and appear to be stable over long periods of time compared with spermatozoa from other vertebrate species.

Nonhuman organisms are being exposed to ionizing radiations at radionuclide-contaminated sites around the world. Direct methods are seldom available for measuring biologically relevant doses received by these organisms. Here we extend biological dosimetry techniques, which are much better developed for humans and a few other mammalian species, to a nonmammalian species. Turtles were chosen because a long-lived animal would best serve the need for low-level, chronic exposure conditions. We chose the yellow-bellied slider turtle (Trachemys scripta), which is known to have a maximum life span of at least 22 years. As reported elsewhere, we first isolated an embryonic fibroblast cell line and constructed whole-chromosome-specific DNA libraries for chromosome 1 by microdissection and PCR. A FISH painting probe was prepared and used to establish a dose-response curve for ionizing radiation-induced chromosome interchange aberrations in turtle fibroblasts. This was compared to the dose response for human fibroblasts treated under similar conditions in our laboratory. With respect to induction of chromosome interchange aberrations, human fibroblasts were approximately 1.7 times more sensitive than the T. scripta fibroblasts. To the extent that symmetrical interchanges are persistent over long periods, this approach could eventually provide a measure of the integrated lifetime dose these organisms receive from radionuclides in their environment and give a measure of the extent of relevant genetic damage over that time.

Optimization of culture techniques for turtle and other reptilian lymphocytes is essential for facilitating cytogenetic and immunologic research for these animals. We examined a variety of conditions and parameters relevant to turtle lymphocyte culture including: different mitogenic agents, alone and in combination; lymphocyte separation protocols; culture volume; time required to stimulate lymphocytes to mitosis; importance of humidity and gas exchange in culture incubation; suitability of different culture media; effects of varying serum concentrations; ability of interleukin-2 (IL-2) to stimulate lymphocyte growth and prevent apoptosis; and feasibility of inducing premature chromosome condensation. The best conditions of those we studied for obtaining mitotic cells were (1) the combined use of phytohemagglutinin-M form (2%) and lipopolysaccharides (0.55 microg/ml), (2) the use of 5% autologous turtle serum (as opposed to fetal bovine serum), and (3) collection of mitotic cells around 96 hours after mitogenic stimulation. Human, recombinant IL-2 did not increase the fraction of lymphocytes in mitosis over the range of concentrations tested and calyculin A was ineffective at inducing premature chromosome condensation in turtle lymphocytes over the range of concentrations tested. This test regime provides a guideline for determination of appropriate lymphocyte culture conditions in turtles and other reptiles.

In many organisms, body size is positively correlated with traits that are presumably related to fitness. If directional selection frequently favors larger offspring (the "bigger is better" hypothesis), the results of such selection should be detectable with field experiments. We tested the "bigger is better" hypothesis in hatchling snapping turtles (Chelydra serpentina) by conducting one long-term and three short-term experiments on the University of Michigan E.S. George Reserve in southeastern Michigan. In the fall of 1995 and 1996, we released hatchlings at artificial nests separated from the nearest wetland by fences. We recorded the proportion of hatchlings recaptured, the time it took hatchlings to move to fences from artificial nests 45, 55, and 80 m away, and dispersion along the fence. We determined whether the response variables and probability of recapture at fences were associated with hatchling body size. During 1995, average travel times of hatchlings from the experimental nests were not related to distance from the fence; however, time to recapture was positively correlated with dispersion from the zero point on the fence, and the maximum time to reach the fence was almost twice as long for hatchlings from the 80-m nest compared to those from the 45-m nest. Sixty-seven percent of the hatchlings reached the fence and the proportions doing so from each nest were not different. Body size was not significantly related to probability of recapture in either of the 1995 experiments. In 1996, 59% of released hatchlings were recaptured. Time to recapture was not related to dispersion from the zero point or to body size. Cubic spline analysis suggested stabilizing selection on body size. We also conducted a set of long-term hatchling release experiments between 1980-1993 to compare the survival of hatchlings released at nest sites to that of hatchlings released directly into marshes, and we looked for relationships between survivorship and hatchling body size. During 7 years in which more than 30 hatchlings were released, 413 hatchlings were released directly into the marsh and 262 were released at nests: their probability of survival did not differ. Over all years, for both release groups combined and for each group separately, survival was not related to body size. In 1983 alone, survival was also not related to body size for either group or for both groups combined. In our three short-term experiments and one long-term experiment, we found no evidence to support the "bigger is better" hypothesis. When selection on body size did occur, selection was stabilizing, not directional for larger size.

To assess the responsiveness of the interrenal axis to stress, we injected toads exposed to coal combustion wastes and toads from an unpolluted reference site with adrenocorticotropic hormone (ACTH), as well as the vehicle alone (saline). Initial circulating levels of corticosterone in toads captured at the polluted area were significantly higher than levels in toads from the reference site. Corticosterone levels in toads from the polluted site remained high even after 2 weeks of laboratory acclimation and injection with saline. The results may suggest disruption of hepatic enzymes responsible for the metabolic clearance of steroid hormones. Injection of toads from the polluted site with ACTH had no effect on plasma corticosterone levels, whereas a similar treatment of toads from the reference site stimulated a marked increase in corticosterone. Our study provides evidence that toads exposed to coal combustion wastes may be less efficient at responding to additional environmental stressors.

Relative to small embryos, large embryos may have longer developmental periods and, subsequently, relatively greater maintenance budgets. Because of the potentially increased metabolic costs of maintaining large embryos for long embryonic periods, Salthe and Mecham (1974) suggested that as ovum size increases among amphibians, ovum lipids (the primary stored metabolic energy reserves) should increase at a proportionally greater rate. To test Salthe and Mecham's hypothesis, we quantified egg lipids for 13 amphibian species from the southeastern United States. As ovum size increased among species, total, nonpolar, and polar lipids increased at rates uniform with or relatively lower than rates of increase in ovum size, in contrast to the hypothesis of Salthe and Mecham. However, variation in ovum lipids among species may be related to differences in breeding biology. Our results indicate that the amount of lipids allocated to ova do not merely depend on ovum size, but rather on the selective environments of the embryo and neonate.

The oviducts of two species of freshwater turtles were examined throughout the year to determine the distribution of sperm within the oviduct, the locations of stored sperm, and the onset of mating. Sperm were stored in undifferentiated tubules and ducts of submucosal glands at the periphery of large glandular segments of the albumen and uterine regions of the oviduct. Sperm transfer to females commenced in October in Sternotherus odoratus and in January in Trachemys scripta. The role of sperm storage in the annual reproductive cycle of freshwater turtles is evaluated in light of the onset of sperm transfer to females and the timing of ovulation and oviposition. Stored sperm are most likely to be utilized in the fertilization of eggs ovulated in the second and subsequent egg clutches.

A number of recent studies have linked developmental, physiological, and behavioral abnormalities in amphibians to coal combustion wastes (coal ash). Few studies, however, have determined trace element concentrations in amphibians exposed to coal ash. In the current study we compare total body concentrations of 20 trace elements in adult southern toads, Bufo terrestris, inhabiting coal ash settling basins with toads that were not exposed to the combustion wastes (reference). In addition, we document the accumulation of trace elements in toads transplanted from reference sites to field enclosures in an ash settling basin for 7 and 12 weeks. Arsenic, selenium, and vanadium levels were significantly elevated in toads captured at the ash-contaminated site in comparison to toads from the reference site. All three of these elements were also significantly elevated in toads exposed to the contaminated habitat for only 7 weeks. Our study suggests that adult anurans can bioaccumulate particularly high levels of selenium and may be useful bioindicators in agricultural and coal ash-impacted habitats.

We measured egg components and pre-ovulatory parental investment in kinosternid turtles (Kinosternon baurii, Kinosternon subrubrum, Sternotherus minor, and Sternotherus odoratus) from the southeastern USA. Allocation patterns were determined by comparing lipid content of eggs and hatchlings, to determine whether females of species with hatchlings that exhibit a delayed nest-emergence strategy: (1) allocate higher proportions of energy storage lipids to eggs, (2) produce hatchlings with higher levels of storage lipids, and (3) have higher levels of pre-ovulatory parental investment in comparison to species whose hatchlings exhibit immediate emergence. Whereas total non-polar lipid (NPL) proportions by dry mass of eggs varied significantly among species, NPL proportions of hatchlings were not significantly different. Pre-ovulatory parental investment in care (proportion of hatchling NPL to egg NPL) was 40, 50, and 55% for K. subrubrum, S. minor, and S. odoratus, respectively. Lipid class composition of eggs and hatchlings was studied to distinguish lipids allocated for energy storage from those allocated to other functions. For both eggs and hatchlings, individual lipid classes (triacylglycerol, triacylglycerol fatty acid, cholesterol, cholesterol ester, and phospholipid) as proportions of total lipid, were similar among species. The major lipid class component of eggs and hatchlings of all species was triacylglycerol (> 83%), an energy storage lipid. Substantial changes in lipid classes during embryogenesis were similar among species and included: (1) depletion of triacylglycerol, (2) increase in cholesterol esters, and (3) changes in phospholipid composition. Incubation time varied significantly among species, and appeared to be responsible for differential energy utilization during embryogenesis. Our results are inconsistent with the previously observed pattern that hatchlings exhibiting a delayed nest-emergence strategy are allocated higher proportions of energy storage lipids than those that exhibit immediate emergence. However, because the species that overwinters in the nest (K. subrubrum) hatches approximately 40 days later than the species that typically does not (S. odoratus), hatchling K. subrubrum may contain higher non-polar lipid proportions than hatchling S. odoratus during similar winter time periods. Kinosternid hatchlings contain enough stored lipids to support basal maintenance costs for substantial time periods. We suggest that such reserves may be critical to hatchling survival during a period of negative energy balance, regardless of nest emergence strategy.

We investigated the relationship between maintenance costs (standard metabolic rates, measured as O2 consumption at rest) in tadpoles of the bullfrog, Rana catesbeiana, and exposure to contaminants in a coal ash-polluted habitat (characterized by a variety of trace elements). We compared metabolic rates of tadpoles collected from the polluted site with those from an unpolluted reference pond. Tadpoles collected in the polluted site had 40%-97% higher standard metabolic rates than those collected from the reference pond. We also reciprocally transplanted eggs of the bullfrog between the polluted site and another reference pond and compared standard metabolic rates of tadpoles at 25 and 80 d posthatching. Metabolic rates of tadpoles raised in the polluted site were from 39% to 175% higher than those raised in a reference pond, depending on tadpole age and temperature at which metabolic rates were measured. There were no effects of site of origin of the eggs (polluted or unpolluted) on metabolic rates. Survival to hatching did not differ between sites, although survival to the end of the experiment (80 d posthatching) was lower in the polluted area than in the reference site. Surviving tadpoles were larger in wet body mass in the polluted site than in the reference pond, possibly due to lower survival in the former, but there was no relationship between survival and metabolic rate. It is clear that some feature of the polluted habitat was responsible for causing substantial elevation of standard metabolic rates of tadpoles. We hypothesize that the mixture of trace elements present in sediment and water in the polluted site was responsible for the observed physiological differences.

This study describes an interrenal stress response in adult toads, Bufo terrestris, after exposure to coal combustion waste (characterized by a variety of trace elements). In the first portion of this study, free-ranging male toads captured at the coal ash polluted site exhibited significantly higher circulating levels of corticosterone (B) in both June/July and August than conspecifics captured at uncontaminated sites. In addition, both calling and noncalling males from the polluted site had higher B levels than conspecifics engaged in the same behaviors at reference sites. Testosterone levels were elevated in toads from the polluted site, regardless of capture month or behavioral state, suggesting altered androgen production, utilization, and/or clearance. In the second portion of this study, male toads from reference sites were transplanted to enclosures at the polluted site or an uncontaminated site. Toads held at the polluted site exhibited significant increases in B after 10 days of exposure compared to toads held at the reference site. B levels remained significantly elevated in toads transplanted to the polluted site after 12 weeks. We hypothesize that high concentrations of various trace elements in the polluted site are responsible for these hormonal responses., (Copyright 1997 Academic Press. Copyright 1997 Academic Press)

/ We examined data relative to species abundance, distribution, anddiversity patterns of reptiles and amphibians to determine how perceptionschange over time and with level of sampling effort. Location data werecompiled on more than one million individual captures or observations of 98species during a 44-year study period on the US Department of Energy's(DOE) Savannah River Site National Environmental Research Park (SRS-NERP) inSouth Carolina. We suggest that perceptions of herpetofaunal speciesdiversity are strongly dependent on level of effort and that land managementdecisions based on short-term data bases for some faunal groups could resultin serious errors in environmental management. We provide evidence thatacquiring information on biodiversity distribution patterns is compatiblewith multiyear spatially extensive research programs and also provide aperspective of what might be achieved if long-term, coordinated researchefforts were instituted nationwide.To conduct biotic surveys on government-managed lands, we recommend revisionsin the methods used by government agencies to acquire and report biodiversitydata. We suggest that government and industry employees engaged inbiodiversity survey efforts develop proficiency in field identification forone or more major taxonomic groups and be encouraged to measure the status ofpopulations quantitatively with consistent and reliable methodologies. Wealso suggest that widespread academic cooperation in the dissemination ofinformation on regional patterns of biodiversity could result byestablishment of a peer-reviewed, scientifically rigorous journal concernedwith status and trends of the biota of the United States. KEY WORDS: Abundance; Amphibian; Biodiversity; Distribution; Landmanagement; Reptile

Long-term ecological data were used to evaluate the relative importance of movements, breeding structure, and reproductive ecological factors to the degree of spatial and age-specific variation in genetic characteristics of painted turtles (Chrysemys picta) on the E. S. George Reserve in southeastern Michigan. Estimates of the degree of spatial genetic structuring were based on the proportion of total genotypic variance partitioned within and between subpopulations (inferred from hierarchical F-statistics based on variation at 18 protein loci), and in terms of gene correlations (co-ancestry among individuals derived from reproductive data on full-sib families of females nesting at specific nesting areas). Little variation in allele frequency was observed among turtles from different marshes (F mt = 0.003), though significant variation was observed among turtles from different nesting areas associated with each marsh (F nm = 0.046). Gene correlations among individuals within nesting areas varied greatly over years (0.032-0.171; mean = 0.069) and were negatively correlated to the proportion of females that successfully nested during each year. General concordance between independent estimates of genotypic correlations (i.e., F nm derived from protein electrophoretic variation vs. mean co-ancestry) suggests that allozyme data, when collected over spatial scales consistent with species behavioral characteristics and reproductive ecology, may accurately reflect the apportionment of gene diversity within and among subpopulations. The magnitude and patterning of allelic variation among nesting areas and individuals appears to be primarily a function of gametic correlations among members of full-sib families, irrespective of the degree of gene flow or female nesting-site fidelity. Comparisons of genetic characteristics among 11 cohorts (1974-1984) revealed that heterozygosity (H) and inbreeding coefficients (F) varied greatly. Cohort estimates of H and F were correlated to female nesting success and to estimates of co-ancestry for the same years. Results clearly reflect the concomitant importance of ecological factors (principally the proportion of the female population that successfully produce offspring during each year) in determining the magnitude and patterning of gene correlations within and among groups, and to the genotypic composition of offspring born during each year., (© 1993 The Society for the Study of Evolution.)

Some freshwater turtles appear unable to produce eggs large enough to achieve the balance between size and number of eggs predicted by optimal egg size theory. We present evidence that pelvic girdle structure constrains egg size and thus offspring size in females of smaller-bodied species (Chrysemys picta and Deirochelys reticularia). The constraint is demonstrated by the correspondence of slopes of the increase of the pelvic aperture and egg width with increasing body size. This constraint appears to be relaxed in a larger-bodied species (Pseudemys scripta), in which the increase in pelvic aperture relative to body size is greater than the increase in egg width. This type of structural constraint on a reproductive trait should not occur unless there is strong selection on pelvic architecture for other functions, such as locomotion, support, and limb retraction, that prevent expansion of the pelvic aperture. Although other explanations may exist for other groups of organisms that appear to vary egg size, the large variation in egg size associated with body size in some species of turtles can be reconciled with optimal egg size theory only if a pelvic constraint model is accepted.

Plasma levels of testosterone (T), thyroxine (T4) and follicle-stimulating hormone (FSH) were measured in a field population of the painted turtle, Chrysemys picta, in Michigan. All three hormones showed pronounced seasonality; plasma luteinizing hormone (LH) remained nondetectable. Plasma T and FSH concentrations were highly correlated and exhibited biphasic cycles with peaks in spring and fall, whereas T4 showed a single peak in summer, coincident with the nadir in T. Hormone levels were minimal on the day that animals emerged from hibernation at the end of March; T and FSH then increased rapidly (in ca. 1 week after emergence) to a transient peak that persisted for only ca. 2 weeks. Plasma T and FSH rose again in September before the onset of hibernation and shortly after the late summer peak in spermatogenetic recrudescence. Plasma T4 increased more gradually after emergence in spring and did not peak for 1-2 months, coincident with the nadir in T. The profile in plasma FSH is consistent with the view that this gonadotropin may regulate androgen secretion but it did not show the expected relationship to the spermatogenetic cycle. Separate studies indicated that the turtles are sensitive to capture and handling; hormone levels (especially androgen) fall precipitously within a day of capture. These "stress" effects may account for discrepancies with previous studies of plasma T in this species.

Coleonyx variegatus is adapted to readily sacrifice its tail to predators. This adaptation is associated with characteristic tail behavior and rapid tail regeneration. There is no facultative metabolic increase associated with tail regeneration, and energy normally allocated to body growth and maintenance is diverted to tail regeneration. This supports the contention that tail behavior, autotomy, and rapid regeneration evolved as mechanisms promoting survival in terms of predator escape.