Multiple environmental stressors induce complex transcriptomic responses indicative of phenotypic outcomes in Western fence lizard.

BACKGROUND: The health and resilience of species in natural environments is increasingly challenged by complex anthropogenic stressor combinations including climate change, habitat encroachment, and chemical contamination. To better understand impacts of these stressors we examined the individual- and combined-stressor impacts of malaria infection, food limitation, and 2,4,6-trinitrotoluene (TNT) exposures on gene expression in livers of Western fence lizards (WFL, Sceloporus occidentalis) using custom WFL transcriptome-based microarrays. RESULTS: Computational analysis including annotation enrichment and correlation analysis identified putative functional mechanisms linking transcript expression and toxicological phenotypes. TNT exposure increased transcript expression for genes involved in erythropoiesis, potentially in response to TNT-induced anemia and/or methemoglobinemia and caused dose-specific effects on genes involved in lipid and overall energy metabolism consistent with a hormesis response of growth stimulation at low doses and adverse decreases in lizard growth at high doses. Functional enrichment results were indicative of inhibited potential for lipid mobilization and catabolism in TNT exposures which corresponded with increased inguinal fat weights and was suggestive of a decreased overall energy budget. Malaria infection elicited enriched expression of multiple immune-related functions likely corresponding to increased white blood cell (WBC) counts. Food limitation alone enriched functions related to cellular energy production and decreased expression of immune responses consistent with a decrease in WBC levels. CONCLUSIONS: Despite these findings, the lizards demonstrated immune resilience to malaria infection under food limitation with transcriptional results indicating a fully competent immune response to malaria, even under bio-energetic constraints. Interestingly, both TNT and malaria individually increased transcriptional expression of immune-related genes and increased overall WBC concentrations in blood; responses that were retained in the TNT x malaria combined exposure. The results demonstrate complex and sometimes unexpected responses to multiple stressors where the lizards displayed remarkable resiliency to the stressor combinations investigated.

Multiple environmental stressors elicit complex interactive effects in the western fence lizard (Sceloporus occidentalis).

Evaluation of multiple-stressor effects stemming from habitat degradation, climate change, and exposure to chemical contaminants is crucial for addressing challenges to ecological and environmental health. To assess the effects of multiple stressors in an understudied taxon, the western fence lizard (Sceloporus occidentalis) was used to characterize the individual and combined effects of food limitation, exposure to the munitions constituent 2,4,6-trinitrotoluene (TNT), and Plasmodium mexicanum (lizard malaria) infection. Three experimental assays were conducted including: Experiment I--TNT × Food Limitation, Experiment II--Food Limitation × Malaria Infection, and Experiment III--TNT × Malaria Infection. All experiments had a 30 day duration, the malaria treatment included infected and non infected control lizards, food limitation treatments included an ad libitum control and at least one reduced food ration and TNT exposures consisting of daily oral doses of corn oil control or a corn oil-TNT suspension at 5, 10, 20, 40 mg/kg/day. The individual stressors caused a variety of effects including: reduced feeding, reduced testes mass, anemia, increased white blood cell (WBC) concentrations and increased mass of liver, kidney and spleen in TNT exposures; reduced cholesterol, WBC concentrations and whole body, testes and inguinal fat weights given food limitation; and increased WBC concentrations and spleen weights as well as decreased cholesterol and testes mass in malaria infected lizards. Additive and interactive effects were found among certain stressor combinations including elimination of TNT-induced hormesis for growth under food limitation. Ultimately, our study indicates the potential for effects modulation when environmental stressors are combined.

Toxicity of octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) in three vertebrate species.

The explosive, octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine or high-melting explosive (HMX), has been found in soils in areas used for testing and training by the military. Many of these areas contain habitat for valued wildlife species. In an effort to better understand the environmental consequences from exposure, a reptilian (western fence lizard [Sceloporus occidentalis]), an amphibian (red-backed salamander [Plethodon cinereus]), and a mammalian species (rabbit [Oryctolagus cuniculus]) were exposed to HMX under controlled laboratory conditions. Lizards and rabbits were exposed to HMX by way of corn oil through gavage, and salamanders were exposed to HMX in soil. Two deaths occurred from acute oral exposures to lizards to 5000 mg HMX/kg BW. Histological and gross pathologic assessment suggested gut impaction as a possible cause of death. Salamanders exposed to concentrations of HMX in soil < or = 1970 mg HMX/kg soil for 10 days did not show adverse effects. Rabbits, however, showed neurologic effects manifested as hyperkinetic events with convulsions at > 24 h after oral exposures. An LD(50) for rabbits was calculated as 93 mg/kg (95% confidence interval 76-117). A subacute 14-day testing regime found a lowest observed effect level of 10 mg/kg-d and a no observed adverse effect level of 5 mg/kg-d based on hyperkinesia and seizure incidence, although changes suggesting functional hepatic alterations were also found. These data suggest that physiologic differences between species, particularly in gastrointestinal structure and function, can affect the absorption of HMX and hence lead to marked differences in toxicity from exposure to the same compound.

Soil ingestion may be an important route for the uptake of contaminants by some reptiles.

Some species of reptiles regularly ingest soil in the wild. Therefore, we evaluated the importance of soil ingestion as a route for the uptake of contaminants in lizards. We used sand as a substitute for soil during the present study. Different groups of leopard geckos (Eublepharis macularius) were provided with a control and five sodium selenite-spiked sand mixtures during a 28-d study. Twenty lizards were assigned to a control group and to each of five selenium-spiked sand mixtures that consisted of nominal selenium (Se) concentrations of 0.05, 0.46, 4.57, 11.41, and 22.83 mg Se/kg dry sand. Leopard geckos readily ingested the Se-spiked sand. We observed concentration-related effects in several endpoints. Overall growth in body mass was the most sensitive endpoint and was significantly (p < 0.05) lower in lizards that ingested the 4.57, 11.41, and 22.83 mg Se/kg sand mixtures compared to controls. Growth in snout-vent length, mean daily food ingestion, and food conversion efficiency were less sensitive and were significantly (p < 0.05) lower in lizards that ingested the 11.41 and 22.83 mg Se/kg sand mixtures compared to controls. Although our results are based on nominal amounts of Se ingested, leopard geckos appear to be as sensitive to sodium selenite as birds and mammals. The present study suggests that ingestion of soil could be an important potential route for the uptake of soil contaminants in some reptiles and should be evaluated in ecotoxicological studies and risk analyses of reptiles.

Toxic effects of oral hexahydro-1,3,5-trinitro-1,3,5-triazine in the western fence lizard (Sceloporus occidentalis).

Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) has been widely used as an explosive in munition formulations, resulting in contamination of wildlife habitat on military installations. To estimate health effects for reptilian species, acute, subacute, and subchronic oral toxicity studies were conducted using the Western fence lizard (Sceloporus occidentalis). Estimated oral median lethal doses were 72 (95% confidence interval [CI], 49-106) mg/kg body weight (slope, 3.754) for males and 88 (95% CI, 65-119) mg/kg (slope, 4.525) for females. Toxicity from RDX suggested the neurological system as the critical target tissue. A 14-d subacute study followed with males dosed orally with RDX (corn oil) at 0, 10, 20, 25, 30, 45, and 60 mg/kg/d. Signs of toxicity frequently included a characteristic body posture. A significant dose-survival relationship was seen over the range of doses, with a significant decrease in survival at 20 mg/kg/d. Males in the 60-d subchronic study were dosed at 0, 1, 2.5, 5, 8, and 11 mg/kg/d, and signs of toxicity included lethargy, cachexia, and anorexia. Survival was decreased at 8 and 11 mg/kg/d. Reduced growth rate and food consumption occurred at 5 mg/kg/d. Brain tissue was assayed for RDX when seizures were observed at a residue concentration of at least 18 microg/g. No abnormalities were observed in the hematologic indices, whereas plasma proteins were reduced. Hepatic enlargement and decreased testes mass occurred at 8 and 11 mg/kg/d. Plasma testosterone concentrations, sperm counts, and motility measures were variable for all treatment levels. Based on survival, growth rate, food intake, and testes to brain weight ratios, these data suggest a lowest-observed-adverse effect level of 5 mg/kg/d and a no-observed-adverse effect level of 2.5 mg/kg/d.

Effects of repeated exposure to malathion on growth, food consumption, and locomotor performance of the western fence lizard (Sceloporus occidentalis).

Effects of repeated pollutant exposure on growth, locomotor performance, and behavior have rarely been evaluated in reptiles. We administered three doses of malathion (2.0, 20, or 100mg/kg body weight) to western fence lizards (Sceloporus occidentalis) over an 81day period. Eight and 23% mortality occurred at 20 and 100mg/kg (p=0.079) and 85% of lizards in the 100mg/kg group exhibited clinical symptoms of poisoning. Growth, food consumption, body condition index, and terrestrial locomotor performance were not significantly influenced by malathion. However, arboreal sprint velocity was significantly reduced in lizards receiving 100mg/kg. Fifty percent of lizards in the 100mg/kg group also refused to sprint in the arboreal setting (p=0.085). Based on these results, arboreal locomotor performance was the most sensitive metric of exposure we evaluated. Further study of compounds such as malathion is warranted due to highly variable application rates and exposure scenarios.

Dose-related effects following oral exposure of 2,4-dinitrotoluene on the Western fence lizard, Sceloporus occidentalis.

2,4-dintitrotoluene (2,4-DNT) is an explosive frequently found in the soil of military installations. Because reptiles can be common on these sites, ecological risk assessments for compounds such as 2,4-DNT could be improved with toxicity data specific to reptiles. Western fence lizards, Sceloporus occidentalis, were used to develop a laboratory toxicity model for reptiles. A hierarchical approach was used; acute to subchronic studies were conducted to provide toxicity data relevant to short- and long-term exposures. First, a modified median lethal dose (LD50) study was conducted on male and female lizards using a stage-wise probit model. The LD50 was 577 mg/kg for female and 380 mg/kg for male lizards. Subsequently, a subacute experiment was conducted to further assess 2,4-DNT toxicity to male lizards and to define exposure levels for a longer term, subchronic study. The subchronic study was conducted for 60 consecutive days; male lizards were exposed to 0, 9, 15, 25, 42, 70 mg/kg/d. Dose-dependent mortality was observed in the three highest dose groups (25, 42, and 70 mg/kg/d); all other animals survived the study duration. Benchmark dose model calculations based on mortality indicated a 5% effect level of 15.8 mg/kg/d. At study termination, a gross necropsy was performed, organ weights were taken, and blood was collected for clinical and hematological analysis. Body weight, kidney weight, food consumption, postdose observations, and blood chemistries all were found to be significantly different from controls at doses above 9 mg/kg/d. Also, preliminary results suggest behavioral observations, and reduced food consumption may be a sensitive indicator of toxicity. The present study indicates Sceloporus occidentalis is suitable for evaluating toxicity of compounds to reptilian species.

Toxicity of oral exposure to 2,4,6-trinitrotoluene in the western fence lizard (Sceloporus occidentalis).

Contamination of the soil with the explosive 2,4,6-trinitrotoluene (TNT) has been found at military sites, many of which are habitats used by reptiles. To provide data useful in assessing ecological risk for reptilian species, acute, subacute, and subchronic oral toxicity studies were conducted with the western fence lizard (Sceloporus occidentalis). Oral median lethal dose (LD50) values for TNT in corn oil were 1,038 and 1,579 mg/kg of body weight for male and female lizards, respectively. Overt signs of toxicity included chromaturia, abdominal enlargement, and tremors. A 14-d subacute study followed in which male lizards were orally dosed with TNT (corn oil) at 0, 33, 66, 132, 263, 525, and 1,050 mg/kg of body weight each day. Clinical signs of toxicity, while similar to the LD50 study, were more subtle and noted in lizards receiving TNT amounts of at least 66 mg/kg/d. Chromaturia was an early consistent sign, often preceding the onset of adverse effects. Male lizards in the 60-d subchronic study were dosed at 0, 3, 15, 25, 35, and 45 mg/kg/d with nearly complete survival (>90%) for lizards in all treatments. Changes in food consumption and body weight were observed at 35 and 45 mg/kg/d. Alterations in hematological end points; blood chemistries (albumin, total protein, alkaline phosphatase, calcium); kidney, spleen, and liver weights; and adverse histopathology were observed in lizards exposed at 25 to 45 mg/kg/d. Testosterone concentration, sperm count, and motility were variable between treatments. Although not significant, incidences of hypospermia and testicular atrophy were observed in some individuals. Together, these data suggest a lowest-observed-adverse effect level of 25 mg/kg/d and a no-observed-adverse effect level of 15 mg/kg/d in S. occidentalis.

The effects of prey species on food conversion efficiency and growth of an insectivorous lizard.

Little is known about the effects of different prey species on lizard growth. We conducted a 6-week study to determine the relative effects of prey species on growth parameters of hatchling western fence lizards, Sceloporus occidentalis. Lizards were fed house cricket nymphs, Acheta domesticus, or mealworm larvae, Tenebrio molitor. The effects of prey species on growth were determined by measuring prey consumption, gross conversion efficiency of food [gain in mass (g)/food consumed (g)], gain in mass, and gain in snout-vent length. Lizards grew well on both the prey species. However, lizards that fed on crickets consumed a significantly higher percentage of their body mass per day than those fed mealworms. Nevertheless, lizards that consumed mealworms ingested significantly more metabolizable energy, had significantly higher food conversion efficiencies, significantly higher daily gains in mass, and significantly greater total growth in mass than lizards that fed on crickets.

Reproductive and thyroid hormone profiles in captive Western fence lizards (Sceloporus occidentalis) after a period of brumation.

Seasonal fluctuation in serum concentrations of sex steroid (testosterone [T] and 17beta-estradiol [E(2)]) and thyroid (triiodothyronine [T(3)] and thyroxine [T(4)]) hormones was determined in captive Western fence lizards (Sceloporus occidentalis). Samples were collected from male and female breeding pairs weekly for a 4-month period after their emergence from artificial brumation. Circulating levels of E(2) corresponded with the expected vitellogenic and ovulatory cycles in females, and surprisingly, E(2) in males followed a similar pattern, indicating a possible role in breeding behavior. Serum T was elevated in male lizards for the first 6 weeks after emergence from brumation, possibly related to an increase in the onset of active spermatogenesis. Thyroid hormones showed little cyclical activity throughout the breeding period, with the exception of small increases of T(3) at weeks 8 and 16, possibly implying an active role of this hormone with ovulation in females. Overall, these baseline hormone data are not only useful in developing this animal as a laboratory reptile model for assessment of endocrine-mediated toxicity, but also of value for understanding herpetological endocrinology and for application in the conservation of threatened species. Zoo Biol 27:36-48, 2008. (c) 2007 Wiley-Liss, Inc.

Impaired terrestrial and arboreal locomotor performance in the western fence lizard (Sceloporus occidentalis) after exposure to an AChE-inhibiting pesticide.

We examined the effects of a commonly used AChE-inhibiting pesticide on terrestrial and arboreal sprint performance, important traits for predator avoidance and prey capture, in the western fence lizard (Sceloporus occidentalis). Lizards were exposed to carbaryl (2.5, 25, and 250 microg/g) and were raced before and 4, 24, and 96 h after dosing. In the terrestrial setting, exposure to low concentrations of carbaryl had stimulatory effects on performance, but exposure to the highest concentration was inhibitory. No stimulatory effects of carbaryl were noted in the arboreal environment and performance in lizards was reduced after exposure to both the medium and highest dose of carbaryl. Our findings suggest that acute exposure to high concentrations of carbaryl can have important sublethal consequences on fitness-related traits in reptiles and that arboreal locomotor performance is a more sensitive indicator of AChE-inhibiting pesticide poisoning than terrestrial locomotor performance.

Assessment of lead uptake in reptilian prey species.

As part of an investigation determining the trophically available fraction of metals in a model terrestrial food web, i.e., invertebrate prey to Western fence lizards (Sceloporus occidentalis), we evaluated the ability of several invertebrate prey to bioaccumulate lead and to form metals-rich granules, which are hypothesized to be non-available to predators. Crickets (Acheta domestica), tenebroid beetle larvae (Tenebrio molitor), and isopods (Porcellio scaber) were selected as model prey organisms. Lack of standard exposure methodologies for these species has presented a barrier to trophic transfer evaluations, as each species has particular requirements that create challenges for designing exposure conditions. We were able to devise exposure conditions for all three species that allow long-term exposure studies. All prey organisms accumulated lead from contaminated food, and for all species the majority of the accumulated Pb was associated with the exoskeleton (>50%), with metals-rich granules accounting for most of the remaining accumulated lead.

Insights into reptile dermal contaminant exposure: Reptile skin permeability to pesticides.

There is growing interest in improving ecological risk assessment exposure estimation, specifically by incorporating dermal exposure. At the same time, there is a growing interest in amphibians and reptiles as receptors in ecological risk assessment, despite generally receiving less research than more traditional receptors. Previous research has suggested that dermal exposure may be more important than previously considered for reptiles. We measured reptile skin permeability to four pesticides (thiamethoxam, malathion, tebuthiuron, trifluralin) using ventral skin samples. All four pesticides penetrated the skin but generally had low permeability. There was no apparent relationship between physicochemical properties and permeability coefficients. Malathion had a significantly greater permeability rate at all time points compared to the other pesticides. Tebuthiuron had a greater permeability than thiamethoxam. Reptiles and mammals appear to have similar skin permeability suggesting that dermal exposure estimates for mammals may be representative of reptiles.

Direct and indirect effects of petroleum production activities on the western fence lizard (Sceloporus occidentalis) as a surrogate for the dunes sagebrush lizard (Sceloporus arenicolus).

The dunes sagebrush lizard (Sceloporus arenicolus) is a habitat specialist of conservation concern limited to shin oak sand dune systems of New Mexico and Texas (USA). Because much of the dunes sagebrush lizard's habitat occurs in areas of high oil and gas production, there may be direct and indirect effects of these activities. The congeneric Western fence lizard (Sceloporus occidentalis) was used as a surrogate species to determine direct effects of 2 contaminants associated with oil and gas drilling activities in the Permian Basin (NM and TX, USA): herbicide formulations (Krovar and Quest) and hydrogen sulfide gas (H2S). Lizards were exposed to 2 concentrations of H2 S (30 ppm or 90 ppm) and herbicide formulations (1× or 2× label application rate) representing high-end exposure scenarios. Sublethal behavioral endpoints were evaluated, including sprint speed and time to prey detection and capture. Neither H2S nor herbicide formulations caused significant behavioral effects compared to controls. To understand potential indirect effects of oil and gas drilling on the prey base, terrestrial invertebrate biomass and order diversity were quantified at impacted sites to compare with nonimpacted sites. A significant decrease in biomass was found at impacted sites, but no significant effects on diversity. The results suggest little risk from direct toxic effects, but the potential for indirect effects should be further explored.

Transfer of selenium from prey to predators in a simulated terrestrial food chain.

Little is known about the accumulation and effects of selenium in reptiles. We developed a simplified laboratory food chain where we fed commercial feed laden with seleno-D,L-methionine (30 microg/g dry mass) to crickets (Acheta domestica) for 5-7 d. Se-enriched crickets (approximately 15 microg/g Se [dry mass]) were fed to juvenile male and female lizards (Sceloporus occidentalis) for 98 d while conspecifics were fed uncontaminated crickets. Lizards fed contaminated prey accumulated Se concentrations ranging from 9.3 (in female carcass) to 14.1 (in female gonad) microg/g compared to <1.5 microg/g in tissues of controls. Female gonad concentrations approached the highest of thresholds for reproductive toxicity in oviparous vertebrates. However, we observed no consistent effect of dietary treatment on sublethal parameters or survival. Our simplified food chain proved to be an ecologically relevant method of exposing lizards to Se, and forms the foundation for future studies on maternal transfer and teratogenicity of Se.

Effect of temperature on toxicity of a natural pyrethrin pesticide to green anole lizards (Anolis carolinensis).

Metabolic rates of reptiles vary with body temperature; therefore, the sensitivity of reptiles to a particular dose level of a pesticide might be expected to vary as well. The purpose of the present study was twofold: To evaluate the effects of temperature on the toxicity to green anole lizards (Anolis carolinensis) of a single concentration of a natural pyrethrin pesticide via percutaneous exposure, and to compare the effects of temperature (20 vs 35 degrees C) on the toxicity of different concentrations of pyrethrins to green anoles. When lizards were exposed to a solution that contained 300 mg/L of pyrethrins, the mortality of lizards maintained at 15 and 20 degrees C was significantly higher (p < 0.01) than the mortality of lizards maintained at 35 and 38 degrees C. In addition, the median lethal concentrations of pyrethrins for lizards maintained at 20 and 35 degrees C were 77.6 and greater than 300 mg/L, respectively. Therefore, temperature clearly influenced the sensitivity of lizards to pyrethrin pesticides.

Improving reptile ecological risk assessment: oral and dermal toxicity of pesticides to a common lizard species (Sceloporus occidentalis).

Reptiles have been understudied in ecotoxicology, which limits consideration in ecological risk assessments. The goals of the present study were 3-fold: to improve oral and dermal dosing methodologies for reptiles, to generate reptile toxicity data for pesticides, and to correlate reptile and avian toxicity. The authors first assessed the toxicity of different dosing vehicles: 100 µL of water, propylene glycol, and acetone were not toxic. The authors then assessed the oral and dermal toxicity of 4 pesticides following the up-and-down procedure. Neither brodifacoum nor chlorothalonil caused mortality at doses ≤ 1750 µg/g. Under the "neat pesticide" oral exposure, endosulfan (median lethal dose [LD50] = 9.8 µg/g) was more toxic than λ-cyhalothrin (LD50 = 916.5 µg/g). Neither chemical was toxic via dermal exposure. An acetone dosing vehicle increased λ-cyhalothrin toxicity (oral LD50 = 9.8 µg/g; dermal LD50 = 17.5 µg/g), but not endosulfan. Finally, changes in dosing method and husbandry significantly increased dermal λ-cyhalothrin LD50s, which highlights the importance of standardized methods. The authors combined data from the present study with other reptile LD50s to correlate with available avian data. When only definitive LD50s were used in the analysis, a strong correlation was found between avian and reptile toxicity. The results suggest it is possible to build predictive relationships between avian and reptile LD50s. More research is needed, however, to understand trends associated with chemical classes and modes of action.

Life-history traits of the lizard Sceloporus undulatus from two populations raised in a common laboratory environment.

Hatchling Sceloporus undulatus elongatus from Washington Co., Utah and S. u. garmani from Woods Co., Oklahoma were raised to maturity and reproduction under identical laboratory conditions with ad libitum food availability. Growth, allometry, age and size of maturity, clutch size and egg mass were compared among lab-raised cohorts from the two populations, among lab-raised and field-caught animals (including their field-caught mothers) and, for growth, with values obtained by previously published field studies on the same or nearby populations. For all traits population differences observed in previous field studies and current field samples resulted from both a plastic response to proximate environmental conditions and intrinsic (possibly genetic) difference. The most plastic traits were growth and age of maturity. Cohorts from both populations expressed the ability to mature in less than 6 months in the laboratory but only the S.u. garmani express early maturity in the field. Allometric differences generated during growth in the lab were not observed in field samples but may reflect an adaptive physiological difference. The least plastic trait was egg mass. The only trait for which the rank order of the difference in the field was reversed in the lab was growth rate. S.u. elongatus grew significantly faster than S.u. garmani in the lab but much slower in the field. The tendency of S.u. garmani females to breed at minimum size of maturity may be greater than that of S.u. elongatus.

Development of a terrestrial vertebrate model for assessing bioavailability of cadmium in the fence lizard (Sceloporus undulatus) and in ovo effects on hatchling size and thyroid function.

In the terrestrial environment, standardized protocols are available for measuring the exposure and effects of contaminants to invertebrates, but none currently exist for vertebrates. In an effort to address this, we proposed that developing lizard embryos may be used as a terrestrial vertebrate model. Lizard eggs may be particularly susceptible to soil contamination and in ovo exposure may affect hatchling size, mortality, as well as thyroid function. Toxicant-induced perturbations of thyroid function resulting from in ovo chemical exposure may result in toxicity during the critical perinatal period in reptiles. Fertilized Eastern fence lizard (Sceloporus undulatus) eggs were placed in cadmium (Cd)-spiked expanded perlite (0, 1.48, 14.8, 148, 1480, 14,800 microg Cd/g, nominal concentrations), artificially incubated at 28 degrees C, and examined daily for mortality. Whole lizard hatchlings as well as failed hatches were homogenized in ethanol and the homogenate was divided for Cd body residue analysis and thyroid hormone (triiodothyronine (T3) and thyroxine (T4)) analyses. Acute mortality was observed in the two highest doses (1480 and 14800 microg Cd/g). Cadmium body residues showed a higher internal concentration with increasing exposure concentration indicating uptake of Cd. There was a decrease in T3:T4 ratio at the highest surviving dose (148 microg Cd/g), however, there were no differences observed in hatchling size measured as weight and snout-vent length, or in whole body thyroid hormone levels. In summary, this study has shown Cd amended to a solid phase representing soil (perlite) can traverse the thin, parchment-like shell membrane of the fence lizard egg and bioaccumulate in lizard embryos. We believe this study is a good first step in investigating and evaluating this species for use as a model.

Evaluation of western fence lizards (Sceloporus occidentalis) and eastern fence lizards (Sceloporus undulatus) as laboratory reptile models for toxicological investigations.

A need is recognized for one or more laboratory reptile models for use in ecotoxicological studies and risk assessments. Maintenance of breeding populations of most reptile species under laboratory conditions is not practical because of their size and slow maturation rate. However, a number of species of spiny lizards (Sceloporus sp.) are small, mature quickly, and reproduce under laboratory conditions. We evaluated three populations of western fence lizards (S. occidentalis) and four populations of eastern fence lizards (S. undulatus) for their performance under laboratory conditions. We reared an F1 generation of each population and compared their performance relative to survival, growth, maturation rate, and reproductive output. A population from the San Joaquin Valley (CA. USA) performed especially well under laboratory conditions and is a good candidate for a laboratory model. We also examined the sensitivity of developing fence lizard embryos to an estrogenic chemical to determine if male secondary sex characteristics were affected. Microinjecting eggs with an estrogenic chemical (17alpha-ethinylestradiol) feminized males and prevented development of embryonic secondary sex characteristics. Therefore, embryonic fence lizards may be useful for studying the effects of endocrine-disrupting chemicals.