Paula F. P. Henry

Fifty-year trends in a box turtle population in Maryland

Abstract A survey conducted in 1995 investigated long term declines reported in a population of box turtles Terrapene carolina monitored each decade since 1945 in bottomland hardwood forest at the Patuxent Wildlife Research Center, Maryland. Methods duplicated past surveys in most respects, but were supplemented by radiotelemetry and a survey of dominant vegetation. Seventy different turtles were found on the 11.8 ha study area, a decline of >75% since peak populations were recorded in 1955. Searchers were less efficient in 1995 than in 1945–1975 for a variety of possible reasons. Among turtles recorded, approximately equal numbers persisted from each of the past five decades, with some individuals surviving >70 years. A sex ratio strongly favoring males was first recorded in 1975 and continued in 1995, but juveniles and subadults were found in greater proportion in 1995 than in any other survey. Six of nine radio-marked turtles left the bottomland study area and migrated to the adjoining bluffs to hibernate, suggesting more extensive movements and perhaps less stable home ranges than formerly thought. Age structure of trees indicated a gradual change to more shade-tolerant species. Examination of rates of change from survey data suggested that major losses probably resulted from changes in hydrology that exacerbated flooding in 1972, with recovery only beginning in 1995 and perhaps limited both by repeated flood events and successional changes in the forest. Slow recovery from losses may indicate that populations of this species would respond poorly to exploitation. ©

A noninvasive, direct real-time PCR method for sex determination in multiple avian species

Polymerase chain reaction (PCR)‐based methods to determine the sex of birds are well established and have seen few modifications since they were first introduced in the 1990s. Although these methods allowed for sex determination in species that were previously difficult to analyse, they were not conducive to high‐throughput analysis because of the laboriousness of DNA extraction and gel electrophoresis. We developed a high‐throughput real‐time PCR‐based method for analysis of sex in birds, which uses noninvasive sample collection and avoids DNA extraction and gel electrophoresis.

The eastern box turtle at the Patuxent Wildlife Research Center 1940s to the present: another view

Several long-term mark recapture studies have been conducted on box turtles (Terrapene c. carolina) providing valuable information on life span, basic demography, home range, and apparent effects of environmental changes on box turtle survival. One of the longest studied populations was first marked in 1942 on the Patuxent Wildlife Research Center in Maryland, and has been surveyed every 10 years until 1995. The age structure and gender ratio of these turtles in the field may support differential habitat use and survival estimates. A few of the turtles first marked during the 1945 study are still observed throughout the Center. Data collected from turtles marked in the more upland habitats during 1985-2002 indicate a younger age class distribution than that observed in the more protected biota of the bottomland floodplain study area. Extrapolating ages of turtles described in data collected throughout the long-term study, it was estimated that turtles, both males and females, can show reproduction-intent behaviors at ages greater than 54 years old. It is suggested that count data collection be continued on a more frequent cycle, extending over a larger part of the Center.

Effect of 17β-trenbolone on male and female reproduction in Japanese quail (Coturnix japonica)

The anabolic steroid 17β trenbolone (17β-TB), a known endocrine disrupting chemical, may influence reproductive functions in avian wildlife. We evaluated the effects of dietary exposure to 17β-TB at 5 and 20 ppm on reproductive functional endpoints in Japanese quail during and after sexual maturation. In the male, 5 and 20 ppm treatments revealed no differences in body and testes weight, testes histology, plasma testosterone concentrations, or size and weight of the foam glands. However, the onset of foam production was significantly earlier (days of age) in the 20 ppm males. In females, dietary 17β-TB at 20 ppm caused a reduction in the number of maturing yellow yolk follicles and overall egg production. Plasma testosterone concentrations were reduced compared to controls. Histology of the oviductal sperm storage tubules was normal in all treatments. The number of sperm holes, sites on the perivitelline layer (PVL) where sperm bound and hydrolyzed a path through the PVL, was significantly greater in the 10th egg laid compared to the 1st egg laid in the 20 ppm treatment. Potential effects, albeit transient, on endpoints associated with male maturation warrant further investigation into the sensitivity of these measures in the event of embryonic and/or trans-generational exposure to 17β-TB.

Bioaccessibility tests accurately estimate bioavailability of lead to quail

Hazards of soil-borne lead (Pb) to wild birds may be more accurately quantified if the bioavailability of that Pb is known. To better understand the bioavailability of Pb to birds, the authors measured blood Pb concentrations in Japanese quail (Coturnix japonica) fed diets containing Pb-contaminated soils. Relative bioavailabilities were expressed by comparison with blood Pb concentrations in quail fed a Pb acetate reference diet. Diets containing soil from 5 Pb-contaminated Superfund sites had relative bioavailabilities from 33% to 63%, with a mean of approximately 50%. Treatment of 2 of the soils with phosphorus (P) significantly reduced the bioavailability of Pb. Bioaccessibility of Pb in the test soils was then measured in 6 in vitro tests and regressed on bioavailability: the relative bioavailability leaching procedure at pH 1.5, the same test conducted at pH 2.5, the Ohio State University in vitro gastrointestinal method, the urban soil bioaccessible lead test, the modified physiologically based extraction test, and the waterfowl physiologically based extraction test. All regressions had positive slopes. Based on criteria of slope and coefficient of determination, the relative bioavailability leaching procedure at pH 2.5 and Ohio State University in vitro gastrointestinal tests performed very well. Speciation by X-ray absorption spectroscopy demonstrated that, on average, most of the Pb in the sampled soils was sorbed to minerals (30%), bound to organic matter (24%), or present as Pb sulfate (18%). Additional Pb was associated with P (chloropyromorphite, hydroxypyromorphite, and tertiary Pb phosphate) and with Pb carbonates, leadhillite (a lead sulfate carbonate hydroxide), and Pb sulfide. The formation of chloropyromorphite reduced the bioavailability of Pb, and the amendment of Pb-contaminated soils with P may be a thermodynamically favored means to sequester Pb. Environ Toxicol Chem 2016;35:2311-2319. Published 2016 Wiley Periodicals Inc. on behalf of SETAC. This article is a US Government work and, as such, is in the public domain in the United States of America.

Spatiotemporal analysis of gene flow in Chesapeake Bay Diamondback Terrapins (Malaclemys terrapin)

There is widespread concern regarding the impacts of anthropogenic activities on connectivity among populations of plants and animals, and understanding how contemporary and historical processes shape metapopulation dynamics is crucial for setting appropriate conservation targets. We used genetic data to identify population clusters and quantify gene flow over historical and contemporary time frames in the Diamondback Terrapin (Malaclemys terrapin). This species has a long and complicated history with humans, including commercial overharvesting and subsequent translocation events during the early twentieth century. Today, terrapins face threats from habitat loss and mortality in fisheries bycatch. To evaluate population structure and gene flow among Diamondback Terrapin populations in the Chesapeake Bay region, we sampled 617 individuals from 15 localities and screened individuals at 12 polymorphic microsatellite loci. Our goals were to demarcate metapopulation structure, quantify genetic diversity, estimate effective population sizes, and document temporal changes in gene flow. We found that terrapins in the Chesapeake Bay region harbour high levels of genetic diversity and form four populations. Effective population sizes were variable. Among most population comparisons, estimates of historical and contemporary terrapin gene flow were generally low (m ≈ 0.01). However, we detected a substantial increase in contemporary gene flow into Chesapeake Bay from populations outside the bay, as well as between two populations within Chesapeake Bay, possibly as a consequence of translocations during the early twentieth century. Our study shows that inferences across multiple time scales are needed to evaluate population connectivity, especially as recent changes may identify threats to population persistence.

Toxicity of Pb‐contaminated soil to Japanese quail (Coturnix japonica) and the use of the blood–dietary Pb slope in risk assessment

This study relates tissue concentrations and toxic effects of Pb in Japanese quail (Coturnix japonica) to the dietary exposure of soil-borne Pb associated with mining and smelting. From 0% to 12% contaminated soil, by weight, was added to 5 experimental diets (0.12 to 382 mg Pb/kg, dry wt) and fed to the quail for 6 weeks. Benchmark doses associated with a 50% reduction in delta-aminolevulinic acid dehydratase activity were 0.62 mg Pb/kg in the blood, dry wt, and 27 mg Pb/kg in the diet. Benchmark doses associated with a 20% increase in the concentration of erythrocyte protoporphyrin were 2.7 mg Pb/kg in the blood and 152 mg Pb/kg in the diet. The quail showed no other signs of toxicity (histopathological lesions, alterations in plasma-testosterone concentration, and body and organ weights). The relation of the blood Pb concentration to the soil Pb concentration was linear, with a slope of 0.013 mg Pb/kg of blood (dry wt) divided by mg Pb/kg of diet. We suggest that this slope is potentially useful in ecological risk assessments on birds in the same way that the intake slope factor is an important parameter in risk assessments of children exposed to Pb. The slope may also be used in a tissue-residue approach as an additional line of evidence in ecological risk assessment, supplementary to an estimate of hazard based on dietary toxicity reference values.

Female hatchling American kestrels have a larger hippocampus than males: A link with sexual size dimorphism?

The brain and underlying cognition may vary adaptively according to an organisms ecology. As with all raptor species, adult American kestrels (Falco sparverius) are sexually dimorphic with females being larger than males. Related to this sexual dimorphism, kestrels display sex differences in hunting and migration, with females ranging more widely than males, suggesting possible sex differences in spatial cognition. However, hippocampus volume, the brain region responsible for spatial cognition, has not been investigated in raptors. Here, we measured hippocampus and telencephalon volumes in American kestrel hatchlings. Female hatchlings had a significantly larger hippocampus relative to the telencephalon and brain weight than males (∼12% larger), although telencephalon volume relative to brain weight and body size was similar between the sexes. The magnitude of this hippocampal sex difference is similar to that reported between male and female polygynous Microtus voles and migratory and non-migratory subspecies of Zonotrichia sparrows. Future research should determine if this sex difference in relative hippocampus volume of hatchling kestrels persists into adulthood and if similar patterns exist in other raptor species, thus potentially linking sex differences in the brain to sex differences of space use of adults in the wild.

Effects on circulating steroid hormones and gene expression along the hypothalamus-pituitary-gonadal axis in adult Japanese quail exposed to 17β-trenbolone across multiple generations

&NA; We investigated the effects of the androgenic growth promoter 17&bgr;‐trenbolone (17&bgr;TB) on adult Japanese quail (Coturnix japonica) exposed across three generations. The F0 generation was exposed after sexual maturity to 0, 1, 5, 10, 20, and 40 ppm through feed. The F1 generation was exposed in ovo by maternal transfer and through feed at the same doses as their parents. The F2 generation was exposed in ovo only. Levels of plasma sex steroids, gonadal Cytochrome P450 aromatase (CYP19A1) mRNA and select brain neuroendocrine peptide mRNAs were measured. In males, testosterone levels did not differ in any generation from those in controls. Estradiol was significantly elevated in 17&bgr;TB treated F0 and F1 males. In F0 and F1 females, testosterone was suppressed by 17&bgr;TB, whereas estradiol was significantly higher at 40 ppm in F0 and at 10 ppm in F1 females. CYP19A1 expression in F1 males and females increased suggesting a compensatory response to the androgenic effects of 17&bgr;TB. Few significant effects were observed in the F2 birds indicating that in ovo exposure had limited effects on the monitored endpoints. Overall, our results confirmed endocrine disrupting effects of dietary 17&bgr;TB in Japanese quail but the response was dependent on sex, developmental stage at initiation of exposure, and dose.

Sex-specific responses in neuroanatomy of hatchling American kestrels in response to embryonic exposure to the flame retardants bis(2-ethylhexyl)-2,3,4,5-tetrabromophthalate and 2-ethylhexyl-2,3,4,5-tetrabromobenzoate: Flame retardants affect neuroanatomy of American kestrels

Bis(2-ethylhexyl)-2,3,4,5-tetrabromophthalate (BEH-TEBP) and 2-ethylhexyl-2,3,4,5-tetrabromobenzoate (EH-TBB), flame retardant components of FireMaster 550® and 600® have been detected in tissues of wild birds. To address the paucity of information regarding potential impacts of flame retardants on the brain, brain volume regions of hatchling American kestrels (Falco sparverius) were evaluated following in ovo injection at embryonic day 5 with safflower oil or to 1 of 3 doses of either BEH-TEBP (12, 60, or 107 ng/g egg) or EH-TBB (11, 55, or 137 ng/g egg). The doses for both chemicals reflected concentrations reported in wild birds. The volumes of the hippocampus and telencephalon and volumetric differences between left and right hemispheres were measured in hatchlings (embryonic day 28). A sex-specific effect of BEH-TEBP on relative hippocampus volume was evident: the hippocampus was significantly enlarged in high-dose females compared to control females but smaller in low-dose females than the other females. There was no significant effect of EH-TBB on hippocampus volume in female kestrel hatchlings or of either chemical in male hatchlings and no effects of these concentrations of EH-TBB or BEH-TEBP on telencephalon volume or the level of symmetry between the hemispheres of the brain. In sum, embryonic exposure of female kestrels to these BEH-TEBP concentrations altered hippocampus volume, having the potential to affect spatial memory relating to ecologically relevant behavior such as prey capture, predator avoidance, and migration. Environ Toxicol Chem 2018;37:3032-3040.