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Fish and aquatic amphibians possess neuromasts on the surface of their body that constitute the lateral line, a sensory system used to detect water displacement. Copper is known to inactivate the neuromast organs of this system. Copper-induced neuromast loss in African clawed frogs, Xenopus laevis, was examined by exposing Nieuwkoop-Faber stage 54-55 larvae to copper concentrations of 0, 100, 200, 300, and 400 µg/L for 96 h, followed by an examination of neuromast counts, staining intensity, and behavioral responses. Neuromasts were counted using a novel imaging method across four different body regions: the whole body, partial body, head, and tail. Neuromast counts showed a decreasing, but nonsignificant, trend across increasing levels of copper exposure. Intensity of neuromast staining showed a stronger concentration-dependent decrease in all four body regions. The decrease in staining intensity, but not neuromast number, may indicate that although neuromasts are still functioning, they have a decreased number of viable hair cells. Potential loss of responsiveness related to neuromast damage was examined via sensitivity to puffs of air at varying distances. We detected little to no difference in response to the air puff stimulus between control tadpoles and tadpoles exposed to 400 µg/L of copper. Neuromasts of X. laevis may be more resistant to copper than those of North American tadpole species, possibly suggesting greater tolerance of the lateral line to environmental stressors in species that maintain this sensory system throughout their lifespan as compared with species that only have the lateral line during the larval period.

Numerous studies have documented that invertebrate pollinator services are critical to the world economy. Factors including habitat loss and agricultural practices, however, threaten pollinator populations. Many counties in the Southern High Plains were identified as at risk for a shortage of pollination service from wild bees. This region also has one of the highest concentrations of Conservation Reserve Program (CRP) contracts in the US. The CRP is the largest, voluntary, private lands conservation program in the US and was targeted as a program to improve pollinator habitat. Our objective was to determine how the predominant land uses in the SHP (native grassland, CRP, and cropland) affect pollinator abundance and species richness, and more specifically if the CRP can provide quality habitat for pollinators. We also examined how the keystone habitat, playa wetlands, embedded within these land uses contribute to pollinator habitat (land type: uplands vs. wetland). We used blue vane traps placed in playa basins and adjacent uplands to determine Hymenoptera abundance and richness from April to October in 2013 and 2014. The CRP had lower abundance than cropland and native grassland, and generally less richness. Uplands and playa wetlands had little difference in Hymenoptera abundance and richness. Patch size negatively influenced abundance but had a positive influence on richness. The interaction of vegetation height and percent bare ground positively influenced abundance in cropland and native grasslands, and positively influenced richness in all land uses. In the CRP, vegetation height negatively influenced Hymenoptera abundance and percent bare ground had a positive influence. The years sampled in this study were during a severe extended drought; therefore, these results may be reflective of poor floral resources. The CRP has potential to create valuable habitat for pollinators if land managers incorporate a diversity of native grasses and native forbs into plantings to enhance pollinator foraging and nesting habitat., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2019 The Authors. Published by Elsevier Ltd.. All rights reserved.)

While puberty is an animal commonality, little is known of its timing or process in crocodylians. Males copulate with an intromittent phallus that has a distinct glans morphology which directly interacts with the female cloaca, putatively effecting effective semen transfer and ultimately increased fecundity. Here we present, during the Morelet's crocodile lifecycle, a well-defined body length (65 cm snout-vent length) inflection point that marks a subsequent increase of phallic glans growth rates. Putatively, this postpubescent growth produces a copulatory-effective phallus. While not as robust of a trend as snout-vent length, this growth inflection concomitantly begins with a body condition index (CI = BM/SVL 3 ) between 2.0 and 2.5 and is most distinct above a CI of 2.5. Also, in males, this 65 cm size threshold also aligns with the initiation of more robust growth in caniniform alveoli associated with prominent maxillary and mandibular teeth. This inflection was not observed in females, thus marking a sexual dimorphism that begins to present with the onset of puberty. This bodily manifestation of puberty other than those changes observed in the reproductive tracts is a novel observation for crocodylians and lays a foundation for further study among species of how changing endocrine signaling within sexually maturing males may also influence a broader range of secondary sex characteristics., (© 2019 Wiley Periodicals, Inc.)

Declining bird populations across the United States have been noted in a number of studies. Although multiple explanations have been proposed as causes of these declines, agricultural intensification has often been suggested as a significant driver of bird population dynamics. Using spatially explicit USDA-NASS Cropland Data Layer, we examined this relationship by comparing bird count data from the Breeding Bird Survey collected between 1995 and 2016 across 13 states in the central United States to corresponding categorical changes in land cover within a 2-km radius of each survey transect. This approach allowed us to compare the slopes of counts for 31 species of birds between grassland- and cropland-dominated landscapes and against increasing levels of cropland (all types combined) and pooled corn and soybean land cover types. Nearly all birds demonstrated significant responses to land cover changes. In all cases, the number of species exhibiting positive or negative responses was comparable, and median differences in percent change per year ranged from -0.5 to 0.7%. Species that responded either positively or negatively did not appear to fall into any particular foraging guild. If changes in agricultural practices are a major cause of declines, we would expect to see it across the spatial scale studied and across the majority of species. While these results do not rule out potential agricultural effects, such as toxicity resulting from pesticide exposure, which may have species-specific or localized effects, a variety of factors related to habitat are likely the most significant contributor overall. Given these results over a large spatial scale basis (multistate) and across numerous bird species, there is not a broad general trend of greater decline in crop-intensive areas. Integr Environ Assess Manag 2018;14:692-702. © 2018 SETAC., (© 2018 SETAC.)

Amphibians are susceptible to exposure from contaminants via multiple pathways. Pyraclostrobin fungicides have been shown to be toxic to terrestrial amphibians at environmentally relevant concentrations; however, these studies did not account for factors that may influence exposure and effects, such as fungicide formulation, age of the individual, exposure route, and physiological state of the individual. We examined Headline® and Headline AMP® fungicide toxicity to adult Anaxyrus cognatus and Anaxyrus woodhousii by direct overspray, as well as acute toxicity of Headline AMP to juvenile A. cognatus through direct overspray, previously exposed soils, and diet. We also assessed effects of hydration state on fungicide toxicity in juvenile A. cognatus and sublethal effects of fungicide exposure on prey-orientation ability of juvenile A. cognatus. Neither formulation of Headline caused mortality of adult A. cognatus and A. woodhousii at up to 5 times the maximum label rate in North American corn (1052 and 879 mL formulation/ha for Headline AMP and Headline fungicides, respectively, corresponding to 1.52 and 2.2 μg pyraclostrobin/cm 2 , respectively). Exposures of juvenile A. cognatus to Headline AMP via direct overspray and previously exposed soils (2 types) resulted in median lethal rate (LR50) values of 2.4, 3.34, and 3.61 μg pyraclostrobin/cm 2 , respectively. Dietary Headline AMP exposure was not toxic, prey-orientations were not influenced by exposure, and effects were similar between dehydrated and hydrated treatments (LR50 = 2.4 and 2.3 μg pyraclostrobin/cm 2 , respectively). These results, combined with exposure concentrations reported in previous studies, suggest that risk of acute mortality for amphibians in terrestrial environments is low and is dictated by body size and site-specific factors influencing exposure. Environ Toxicol Chem 2017;36:2679-2688. © 2017 SETAC., (© 2017 SETAC.)

Playa wetlands, the dominant hydrological feature of the semi-arid U.S. High Plains providing critical ecosystem services, are being lost and degraded due to anthropogenic alterations of the short-grass prairie landscape. The primary process contributing to the loss of playas is filling of the wetland through accumulation of soil eroded and transported by precipitation from surrounding cultivated watersheds. We evaluated effectiveness of vegetative buffers surrounding playas in removing metals, nutrients, and dissolved/suspended sediments from precipitation runoff. Storm water runoff was collected at 10-m intervals in three buffer types (native grass, fallow cropland, and Conservation Reserve Program). Buffer type differed in plant composition, but not in maximum percent removal of contaminants. Within the initial 60 m from a cultivated field, vegetation buffers of all types removed >50% of all measured contaminants, including 83% of total suspended solids (TSS) and 58% of total dissolved solids (TDS). Buffers removed an average of 70% of P and 78% of N to reduce nutrients entering the playa. Mean maximum percent removal for metals ranged from 56% of Na to 87% of Cr. Maximum removal was typically at 50 m of buffer width. Measures of TSS were correlated with all measures of metals and nutrients except for N, which was correlated with TDS. Any buffer type with >80% vegetation cover and 30-60 m in width would maximize contaminant removal from precipitation runoff while ensuring that playas would continue to function hydrologically to provide ecosystem services. Watershed management to minimize erosion and creations of vegetation buffers could be economical and effective conservation tools for playa wetlands., (Published by Elsevier Ltd.)

Amphibian metamorphosis is complex and larval morphology and physiology are completely restructured during this time. Amphibians that live in unpredictable environments are often exposed to stressors that can directly and indirectly alter physiological systems during development, with subsequent consequences (carryover effects) later in life. In this study, we investigated the effects of water level reduction on development rate, spleen size and cellularity, and examined the role of corticosterone levels in premetamorphic, metamorphic, and postmetamorphic New Mexico spadefoot toads (Spea multiplicata). Based on previous studies, we hypothesized that declining water level would increase tadpole developmental rate, but with the trade-off of increasing corticosterone to a level that would subsequently affect spleen size and cellularity, thus prolonging potential immunological suppression. Declining water levels increased developmental rate by 3 days; however, there were no significant body size effects. Corticosterone (CORT) was negatively correlated with total length, snout vent length, body weight, and spleen weight at metamorphosis, suggesting that size at metamorphosis and the immune system may be affected by excessive CORT levels. When compared to other studies, our results support the view that multiple factors may be acting as stressors in the field affecting amphibian responses, and simple pathways as tested in this study may not adequately represent field conditions., (© 2016 Wiley Periodicals, Inc.)

Prairie potholes are the dominant wetland type in the intensively cultivated northern Great Plains of North America, and thus have the potential to receive pesticide runoff and drift. We examined the presence of pesticides in sediments of 151 wetlands split among the three dominant land use types, Conservation Reserve Program (CRP), cropland, and native prairie, in North and South Dakota in 2011. Herbicides (glyphosate and atrazine) and fungicides were detected regularly, with no insecticide detections. Glyphosate was the most detected pesticide, occurring in 61% of all wetlands, with atrazine in only 8% of wetlands. Pyraclostrobin was one of five fungicides detected, but the only one of significance, being detected in 31% of wetlands. Glyphosate was the only pesticide that differed by land use, with concentrations in cropland over four-times that in either native prairie or CRP, which were equal in concentration and frequency of detection. Despite examining several landscape variables, such as wetland proximity to specific crop types, watershed size, and others, land use was the best variable explaining pesticide concentrations in potholes. CRP ameliorated glyphosate in wetlands at concentrations comparable to native prairie and thereby provides another ecosystem service from this expansive program., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Wetlands in the Rainwater Basin (RWB) of Nebraska are commonly in close proximity to or embedded within row-crop agriculture. Several fungicides and bifenthrin are applied aerially to corn throughout the RWB during tassel stage. Thus, aerial spray drift and runoff may result in pesticide contamination of wetlands. The primary objective of this study was to determine water concentrations of five fungicides and bifenthrin in wetlands located in and near fields during a heavy application period (July 16th-26th 2014) to evaluate the influence of distance from crop field on wetland contamination. In addition, the sampling sites were grouped based upon the type of water body sampled and environmental concentrations were compared to relevant "levels of concern" (LOCs) from invertebrates, fish, and amphibians selected from the literature based upon relevant toxicity data. Bifenthrin was the most frequently detected analyte, appearing in 83% of the samples. The 95th percentile concentrations across all wetlands were found to be 0.07 μg/L for bifenthrin, 0.28 μg/L for pyraclostrobin, 0.28 μg/L for azoxystrobin, and <0.14 μg/L for all other analytes. Analyte concentrations did not differ by wetland type and were not correlated with distance from closest crop. Environmental concentrations of fungicides were lower than LOCs, indicating limited acute toxicity risk. However, bifenthrin concentrations were frequently higher than LOCs for aquatic arthropods. Because the water samples included particulate bound pesticides, further work is needed to determine if this bifenthrin is bioavailable and contamination levels within the sediment., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Previous laboratory studies have suggested that pyraclostrobin-containing fungicide formulations are toxic to amphibians at environmentally relevant concentrations. However, it is unknown if all pyraclostrobin formulations have similar toxicity and if toxicity occurs in different amphibian species. We investigated the acute toxicity of two formulations, Headline(®) fungicide and Headline AMP(®) fungicide, to Blanchard's cricket frogs (Acris blanchardi) based on a direct overspray scenario. In addition, we examined body residues of fungicide active ingredients in A. blanchardi following direct exposure to Headline AMP fungicide. Headline fungicide and Headline AMP fungicide had similar toxicity to A. blanchardi with calculated median lethal doses of 2.1 and 1.7 µg pyraclostrobin/cm(2), respectively, which are similar to the suggested maximum label rate in North American corn (2.2 and 1.52 µg pyraclostrobin/cm(2), respectively). Tissue concentrations of pyraclostrobin were lower than predicted based on full uptake of a direct dose, and did not drop during the first 24 h after exposure. Headline fungicides at corn application rates are acutely toxic to cricket frogs, but acute toxicity in the field will depend on worst-case exposure.

Methyl mercury cation (MeHg(+)) and divalent mercury (Hg(2+)) were quantified in urine, liver, kidney, and brain of prairie voles (Microtus ochrogaster) during a 12 week exposure to aqueous MeHg(+) at concentrations of 10, 100, and 1000 ng MeHg(+)/mL. Aqueous MeHg(+) exposures increased mercury accumulation in tissues of voles from each exposure group. Accumulation was greater within the higher two exposure groups. Similar [Hg(2+)] and [MeHg(+)] were determined within a given organ type before and after 2,3-dimercapto-1-propane sulfonate (DMPS) chelation. Similar correlations were seen for Hg(2+) and MeHg(+) concentrations in pre and post chelation urine. Post chelation urine more reliably predicted mercury species concentrations in tissues than did urine collected before chelation. These data demonstrate the utility of DMPS in noninvasive assessment of wildlife exposure to mercury, which may have utility in evaluating meta-population level exposure to hazardous wastes.

Recent studies have demonstrated that a pyraclostrobin-containing fungicide (Headline(®) Fungicide--Headline(®) Fungicide and Headline AMP(®) Fungicide are registered trademarks of BASF) is toxic to amphibians at environmentally relevant concentrations. However, these studies were performed in a laboratory setting of a worst-case direct exposure in clean media. Interception of spray by the crop canopy and ground cover used by animals for security cover will influence exposure. Thus, risk to amphibians is unclear in an environmentally realistic field environment. We tested exposure and toxicity of Headline AMP(®) Fungicide to amphibians in multiple agricultural habitat scenarios (e.g., within treated crop vs. grassy areas adjacent to crop) and at two rates during routine aerial application. Specifically, we placed Woodhouse's toads (Bufo woodhousii) and Blanchard's cricket frogs (Acris blanchardi) in enclosures located within treated and untreated corn (VT stage, approximate height = 3 m), and in the potential drift area (adjacent to treated corn) during aerial application of Headline AMP Fungicide at either 731 or 1052 ml/ha (70 and 100 % the maximum application rate in corn, respectively). Mean concentrations of pyraclostrobin measured at ground level were ≤19 % of nominal application rate in all areas. Overall, mean mortality of recovered individuals of both species was ≤15 %, and mortality within Headline AMP Fungicide-treated corn (where risk was anticipated to be highest) was <10 %. It is important to understand that application timing, interception by the crop canopy (which varies both within and between crop systems), and timing of amphibian presence in the crop field influences risk of exposure and effects; however, our results demonstrate that amphibians inhabiting VT stage corn during routine aerial application of Headline AMP Fungicide are at low risk for acute mortality, matching existing laboratory results from acute toxicity studies of Headline Fungicide.

With the increased use of fungicides in cultivated regions such as the southern High Plains (SHP), U.S., unintentional runoff and drift as well as direct overspray during aerial application lead to environmental exposures that may influence soil microbial communities and related biogeochemical functioning. Our goal was to examine the effects of two popular fungicides Headline (pyraclostrobin) and Quilt (azoxystrobin/propiconazole) on respiration from soil microbial communities in playa wetlands embedded in cropland and native grassland and their adjacent watersheds. We monitored fungicide effects (at levels of 0, .1×, 1× and 10× the label rate) by measuring respiration from plant matter amended soils collected from 6 cropland and 6 grassland playas and uplands. In addition, differences in microbial community structure among land use types were determined by measuring ergosterol levels in cropland and native grassland playas and uplands. Native grassland playas and their associated watersheds had up to 43% higher soil respiration rates than cropland playas and watersheds, indicating higher soil microbial activity. Application of either fungicide had no effect on soil respiration at any concentration in either land use type or habitat type (playa/watershed). Native grassland playas and watersheds had 3 and 1.6 times higher ergosterol content than cropland playas and watersheds. The lack of soil respiration response to fungicide application does not necessarily suggest that fungicides used in this study do not affect non-target soil microbial communities due to potential compensation by other biota. Future studies should further elucidate existing microorganism communities in playas and their watersheds., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Watershed cultivation and subsequent soil erosion remains the greatest threat to the service provisioning of playa wetlands in the High Plains. The U.S. Department of Agriculture's (USDA) Conservation Reserve Program (CRP) plants perennial vegetation cover on cultivated lands including playa watersheds, and therefore, the program influences sediment deposition and accumulation in playas. Our objective was to measure the effects of the CRP on sediment deposition by comparing sediment depth and present/historic size characteristics in 258 playas among three High-Plains subregions (northern, central, and southern) and the three dominant watershed types: cropland, CRP, and native grassland. Sediment depth and resultant volume loss for CRP playas were 40% and 57% lower than cropland playas, but 68% and 76% greater than playas in native grassland. Playas in CRP had remaining volumes exceeding those of cropland playas. Grassland playas had nearly three times more original playa volume and 122% greater wetland area than CRP playas. Overall, playas were larger in the south than other subregions. Sediment depth was also three times greater in the south than the north, which resulted in southern playas losing twice as much total volume as northern playas. However, the larger southern playas provide more remaining volume per playa than those in other subregions. The results of this study demonstrate the importance of proper watershed management in preserving playa wetland ecosystem service provisioning in the High Plains. Furthermore, we identify regional differences in playas that may influence management decisions and provide valuable insight to conservation practitioners trying to maximize wetland services with limited resources.

Agricultural fungicide application has increased tenfold since 2005 in the United States. Active ingredients and formulations of strobilurin fungicides at environmentally relevant concentrations cause mortality to larval and metamorph amphibians; however, little is known about chronic exposure effects in amphibians. We exposed larval amphibians (Bufo cognatus) throughout metamorphosis to the common fungicide formulations Headline(®), Stratego(®), Quilt(®), and a control to determine effects on development and growth. Formulations were tested at 1.7, 50, and 400 μg/L of the active strobilurin ingredient for Headline(®), Stratego(®), and Quilt(®), respectively. Fungicide exposure did not affect body mass or snout-vent length at metamorphosis. However, exposure to Headline(®) at 1.7 μg/L increased the development rate of tadpoles by approximately 5 days compared to the control, an effect not observed for Stratego(®) and Quilt(®). Stratego(®) also caused approximately 35 % cumulative mortality. Results from the experiment suggest that chronic effects of strobilurin fungicides on development, growth, and mortality to B. cognatus are apparent at environmentally relevant concentrations.

Fungicide application rates on row crop agriculture have increased across the United States, and subsequently, contamination of adjacent wetlands can occur through spray drift or field runoff. To investigate fungicide toxicity, Hyalella azteca amphipods were exposed to 2 fungicide formulations, Headline and Stratego, and their active strobilurin ingredients, pyraclostrobin and trifloxystrobin. Water-only exposures resulted in similar median lethal concentration (LC50; 20-25 µg/L) values for formulations and strobilurin ingredients, suggesting that toxicity is due to strobilurin ingredients. These values were below concentrations that could occur following spray drift over embedded cropland wetlands. When fungicides were added to overlying water of sediment-water microcosms, toxicity was reduced by 500% for Headline and 160% for Stratego, compared with water-only exposures, based on the total amount of fungicide added to the systems. In addition, when fungicides were added to sediment prior to the addition of water, the reduction in toxicity was even greater, with no toxicity occurring at environmentally relevant levels. Differences in toxicity among exposure groups were explained by dissipation from water as toxicity values based on measured water concentrations were within 20% between all systems. The present study reinforces previous studies that Headline and Stratego are toxic to nontarget aquatic organisms. However, the presence of sediment is likely to ameliorate some toxicity of fungicide formulations, especially if spraying occurs prior to wetland inundation., (Copyright © 2013 SETAC.)

The 25000 playa wetlands within the Southern High Plains (SHP) of the United States of America (USA) are the dominant hydrogeomorphic feature in the region, providing habitat for numerous plants and wildlife. The SHP are among the most intensively cultivated regions; there are concerns over the degradation and/or loss of playa wetland habitat. We examined water quality in playa wetlands surrounded by both grassland and agriculture and measured water concentrations of pesticides used on cotton (acephate, trifluralin, malathion, pendimethalin, tribufos, bifenthrin, λ-cyhalothrin, acetamiprid, and thiamethoxam), the dominant crop in the SHP. Pesticides used on cotton were detected in water samples collected from all playas. Precipitation events and the amount of cultivation were related to pesticide concentrations in sediment and water. Our results show that pesticide concentrations were related in some circumstances to time, precipitation, and tilled-index for some but not all pesticides. We further compared measured pesticide concentrations in playas to toxicity benchmarks used by the US EPA in pesticide ecological risk assessments to obtain some insight into the potential for ecological effects. For all pesticides in water, the maximum measured concentrations exceeded at least one toxicity benchmark, while median concentrations did not exceed any benchmarks. This analysis indicates that there is a potential for adverse effects of pesticides to aquatic organisms., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Identifying community assembly filters is a primary ecological aim. The High Plains, a 30 million ha short-grass eco-region, is intensely cultivated. Cultivation disturbance, including plowing and eroded soil deposition down-slope of plowing, influences plant composition in depressional wetlands, such as playas, within croplands. We evaluated influences of wetland cultivation and sediment deposition on plant composition in playas embedded within croplands (46 plowed and 32 unplowed) and native grasslands (79) across 6 High Plains' states. Sediment accumulation ranged from 7 to 78 cm in cropland and 1 to 35 cm in grassland playas. Deeper sediments and plowing each decreased wetland plant richness, 28% and 70% respectively in cropland wetlands. Sediment depth reduced richness 37% in small grasslands playas while it increased richness 22% in larger ones, suggesting moderate disturbance increased richness when there were nearby propagule sources. Sediment depth was unrelated to species richness in plowed wetlands, probably because plowing was a strong disturbance. Plowing removed perennial plants from vegetation communities. Sediment accumulation also influenced species composition in cropland playas, e.g., probability of Eleocharis atropurpurea increased with sediment depth, while probability of Panicum capillare decreased. In grassland playas, observed lighter sediment depths did not influence species composition after accounting for wetland area. Sediment accumulation and plowing shift wetland plant communities toward annual species and decrease habitat connectivity for wetland-dependent organisms in cropland playas over 39,000 and 23,400 ha respectively. Conservation practices lessening sediment accumulation include short-grass buffer strips surrounding wetlands. Further, wetland tillage, allowed under federal agricultural conservation programs, should be eliminated., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Playas are the primary wetland system in the Southern Great Plains (SGP) of North America providing critical stopover habitats for migratory birds in the Western Hemisphere. Collectively, these wetlands form the keystone ecosystem in this region supporting biodiversity for North America and provide habitat for native plants and animals that are essential for maintenance of international biological diversity along with other local and regional ecological services. Large-scale landscape changes in this region, primarily as a result of agriculturally-based anthropogenic impacts, threaten playas with functional loss and physical extinction. These impacts are not considered in current estimates of the number of extant functional playas, leading to biased estimates by groups or agencies extrapolating research or monitoring results, conducting conservation planning, and modeling impacts of future climate change. Using a combination of stochastic and empirical data, we identified impacts to playas and playa watersheds and quantified the extent and rate of these impacts relative to physical loss. Only 0.2% of playas had no wetland or watershed modification and we conservatively estimate that 17% of playas recently existing on the landscape no longer are represented by an apparent depression. With the inclusion of sediment volume estimates where ≥100% of the volume of original differentiated playa soil has been filled, 60% or 16,855 playas have been physically lost from the SGP despite a continued presence of a depression. Data also show that small playas are being lost more rapidly than larger ones; the average size (±SE) of extant playas has increased from 7.5 ± 0.47 to 8.5 ± 0.55 ha. The reduced ecological condition of 95.3% of playas remaining on the landscape exceeds historical predictions of a maximum 85% of playas ever being modified. Given the results of this study these changes can now be accounted for, resulting in more informed management and conservation of playas and bring awareness to the urgency of implementation of effective conservation measures for playas and the species that depend on this keystone ecosystem., (Published by Elsevier Ltd.)

Fungicide applications in the United States have increased tenfold in the last 5 years. Formulations and active ingredients (AIs) have been demonstrated to cause acute mortality to amphibian life stages. However, there has been little to no discrimination between the toxicity of fungicide formulations and their AIs. Therefore, we compared the acute toxicity of the active ingredients and formulations of the fungicides Headline®, Stratego®, and Quilt® using Bufo cognatus tadpoles exposed to four concentrations and a control. All fungicides, including AIs and formulations, demonstrated toxicity to tadpoles, with Headline® and Stratego® causing 100 % mortality at the highest concentrations. Exposure to Quilt® formulation and its AIs resulted in 50-60 % tadpole mortality. Overall, toxicity was comparable between AIs and formulations for all fungicides and concentrations, with the exception of Headline® at 5 μg/L, where formulation exposure resulted in 79 % mortality versus no mortality from exposure to the AI. Results suggest the AIs are responsible for most mortality for Quilt® and Stratego®. Results for Headline® however suggest that although the AI is toxic to tadpoles at environmentally relevant concentrations, adjuvant(s) in the Headline® formulation also contribute to mortality, making it the most toxic of the fungicides studied.

We examined pesticide contamination in sediments from depressional playa wetlands embedded in the three dominant land-use types in the western High Plains and Rainwater Basin of the United States including cropland, perennial grassland enrolled in conservation programs (e.g., Conservation Reserve Program [CRP]), and native grassland or reference condition. Two hundred and sixty four playas, selected from the three land-use types, were sampled from Nebraska and Colorado in the north to Texas and New Mexico in the south. Sediments were examined for most of the commonly used agricultural pesticides. Atrazine, acetochlor, metolachlor, and trifluralin were the most commonly detected pesticides in the northern High Plains and Rainwater Basin. Atrazine, metolachlor, trifluralin, and pendimethalin were the most commonly detected pesticides in the southern High Plains. The top 5-10% of playas contained herbicide concentrations that are high enough to pose a hazard for plants. However, insecticides and fungicides were rarely detected. Pesticide occurrence and concentrations were higher in wetlands surrounded by cropland as compared to native grassland and CRP perennial grasses. The CRP, which is the largest conservation program in the U.S., was protective and had lower pesticide concentrations compared to cropland.

Environmental contamination by energetic compounds is an increasing international concern, although little is known of their accumulation in and affect on wildlife. Reptiles are often good models for contaminants studies due to natural history traits that increase their potential for exposure. We report a study to assess accumulation and effects of octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX, High Melting Explosive) in green anoles (Anolis carolinensis). Acute oral toxicity (LD(50)) was estimated to exceed 2000 mg/kg body weight in adult male and female anoles using a standard up-and-down method. Accumulation of HMX was assessed in adult females via dietary exposure and into eggs by two routes (directly from the soil and via maternal transfer). HMX readily accumulated into adult females in a dose-dependent manner and into eggs following both exposure pathways. However, total HMX in soil-exposed eggs was up to 40-times greater than those exposed via maternal transfer. Although there was a suggestion of an HMX-induced reduction in body weight in adult females, overall there were no effects observed over the 12 week exposure period. The only significant effect on eggs was a 50% reduction in hatching success for eggs exposed to 2000 mg/kg HMX in the soil during incubation. Growth and survival of hatchlings was not affected by HMX exposure. Our results demonstrate that HMX accumulates through the food chain and into eggs from the soil, but likely poses minimal threat to lizards except to hatching success in eggs incubated in soils with HMX levels near maximum environmental concentrations.

This study evaluated the interactions of flight, fasting, and 1,1,1-trichloro-bis(4-chlorophenyl)ethane (p,p'-DDT) loading on residue metabolism and distribution in recently exposed white-crowned sparrows (Zonotrichia leucophrys). Female sparrows were dosed with 5 mg p,p'-DDT per kg body weight over 3 d. Following 1 d of recovery, sparrows were flown in a wind tunnel for up to 140 min, in 15-min blocks. Food was withheld from the start of the flight period until birds were euthanized. DDT, 1,1-dichloro-2,2-bis(4 chlorophenyl)ethane (DDD), and 1,1-dichloro-2,2-bis(4-chlorophenyl)ethylene (DDE) were present in all tissues examined. 1-Chloro-2,2-bis(4-chlorophenyl)ethene (DDµ), 1,1-bis(4-chlorophenyl)ethane (p,p'-DDη), and 2,2-bis(4-chlorophenyl)ethanol (p,p'-DDOH) were not found. Fasting did not significantly affect the rate of residue increase over time in any of the tissues examined. When sparrows flew and fasted simultaneously, fasting seldom contributed to an increase in tissue residues. However, the length of time flown was significantly correlated with increasing toxicant concentrations in the brain, kidney, and liver, effectively demonstrating the potential for brief flights to enhance mobilization of DDT and its metabolites. Dose, flight, and fasting also increased residues in brain tissue. These contaminant redistributions may have important ramifications on the stresses experienced by migratory birds., (Copyright © 2011 SETAC.)

Pesticides have been implicated in widespread amphibian declines. We assessed acute and chronic toxicity of two widely used herbicides to larval New Mexico (Spea multiplicata) and Plains (S. bombifrons) spadefoots from cropland and native grassland playas. Roundup WeatherMAX (WM) toxicity estimates (48- and 216-h LC(50); 48-h LC(1)) for both species were similar to environmental concentrations expected from accidental overspray. Chronic (30-day) exposure to WM at predicted environmental concentrations (2.0 and 2.8 mg glyphosate acid equivalents/L) reduced survival of both species. Ignite 280 SL (IG) toxicity estimates (48-h LC(50) and LC(1)) for both species were above predicted environmental concentrations of 1.0 mg glufosinate/L. Chronic exposure to predicted environmental concentrations of IG did not reduce survival of either species. Toxicity test results suggest that at predicted environmental concentrations IG would not cause extensive mortalities among larval New Mexico and Plains spadefoots. However, WM may cause extensive mortality among larvae of these species., (Copyright 2010 Elsevier Ltd. All rights reserved.)

Mass die-offs of amphibian populations pose a challenging problem for conservation biologists. Ranaviruses often cause systemic infections in amphibians and, in North America, are especially virulent and lethal to larvae and metamorphs. In this paper we describe a novel ranavirus isolate as well as the first recorded occurrence of ranavirus in the southern High Plains of Texas and in associated populations of the spotted chorus frog Pseudacris clarkii. The breeding sites were playas, that is, wetlands that fill via isolated thunderstorms that can occur infrequently; thus, not every playa has water or breeding amphibians annually. We did not detect ranavirus in sympatric anurans, but other reports document ranaviruses in Pseudacris spp. elsewhere. The occurrence of multiple isolates of ranavirus in a number of Pseudacris species suggests that this genus of frogs is highly susceptible to ranaviruses and may experience exceptionally high mortality rates from infection. Thus, the virus may contribute to substantial seasonal population declines and low seasonal recruitment, with negative impacts on populations of breeding adults in successive years.

Pesticide toxicity is often proposed as a contributing factor to the world-wide decline of amphibian populations. We assessed acute toxicity (48 h) of a glufosinate-based herbicide (Ignite 280 SL) and several glyphosate-based herbicide formulations (Roundup WeatherMAX, Roundup Weed and Grass Killer Super Concentrate, Roundup Weed and Grass Killer Ready-To-Use Plus on two species of amphibians housed on soil or moist paper towels. Survival of juvenile Great Plains toads (Bufo cognatus) and New Mexico spadefoots (Spea multiplicata) was reduced by exposure to Roundup Weed and Grass Killer Ready-To-Use Plus on both substrates. Great Plains toad survival was also reduced by exposure to Roundup Weed and Grass Killer Super Concentrate on paper towels. New Mexico spadefoot and Great Plains toad survival was not affected by exposure to the two agricultural herbicides (Roundup WeatherMAX and Ignite 280 SL) on either substrate, suggesting that these herbicides likely do not pose an immediate risk to these species under field conditions.

Microsatellite data were generated from hatchlings collected from ten nests of Morelet's Crocodile (Crocodylus moreletii) from New River Lagoon and Gold Button Lagoon in Belize to test for evidence of multiple paternity. Nine microsatellite loci were genotyped for 188 individuals from the 10 nests, alongside 42 nonhatchlings from Gold Button Lagoon. Then mitochondrial control region sequences were generated for the nonhatchlings and for one individual from each nest to test for presence of C. acutus-like haplotypes. Analyses of five of the nine microsatellite loci revealed evidence that progeny from five of the ten nests were sired by at least two males. These data suggest the presence of multiple paternity as a mating strategy in the true crocodiles. This information may be useful in the application of conservation and management techniques to the 12 species in this genus, most of which are threatened or endangered.

Soil and topical tests were employed to investigate the effect of two N-nitroso metabolites of RDX (hexahydro-1,3,5-trinitro-1,3,5-triazine) on earthworm reproduction. The lowest observed effect concentration (LOEC) for cocoon production and hatching was 50mg/kg for both hexahydro-1-nitroso-3,5-dinitro-1,3,5-triazine (MNX) and hexahydro-1,3,5-trinitroso-1,3,5-triazine (TNX) in soil. MNX and TNX also significantly affected cocoon hatching in soil (p<0.001) and in topical tests (p=0.001). The LOECs for cocoon hatching were 1 and 10mg/kg for MNX and TNX in soil, respectively, and 10mg/L in the topical test. Greater than 100mg/kg MNX and TNX completely inhibited cocoon hatching. In soil, the EC20 values for MNX were 8.7 and 8.8mg/kg for cocoon and juvenile production, respectively, compared to 9.2 and 9.1mg/kg for TNX, respectively. The EC20 values for the total number of cocoon hatchlings were 3.1 and 4.7mg/kg for MNX and TNX, respectively, in soil and 4.5 and 3.1mg/L in the topical test. Both MNX and TNX inhibited cocoon production and hatching, suggesting that they may have a negative affect on soil ecosystems at contaminated sites.

Playa wetlands are critical habitat for wildlife in the Southern High Plains (SHP), a region dominated by agriculture. Little information on pesticide levels exists for playas, and thus we measured organochlorine pesticide concentrations in sediment and amphibians collected from playas in cropland and grassland watersheds. Heptachlor, alpha- and beta-BHC, gamma-chlordane, and dieldrin were detected in sediment and/or tissue samples, typically at or below 1 ng/g, dry weight. However, mean DDT and DDE reached 19.7 and 4.1 ng/g in sediments and 6.3 and 2.4 ng/g in tissues, respectively. Land use did not influence pesticide levels in sediment or amphibians.

A reliable, sensitive, and reproducible method was developed for quantitative determination of nine new generation pesticides currently used in cotton agriculture. Injector temperature significantly affected analyte response as indicated by electron capture detector (ECD) chromatograms. A majority of the analytes had an enhanced response at injector temperatures between 240 and 260 degrees C, especially analytes such as acephate that overall had a poor response on the ECD. The method detection limits (MDLs) were 0.13, 0.05, 0.29, 0.35, 0.08, 0.10, 0.32, 0.05, and 0.59 ng/mL for acephate, trifuralin, malathion, thiamethozam, pendimethalin, DEF6, acetamiprid, brifenthrin, and lambda-cyhalothrin. This study provides a precision (0.17-13.1%), accuracy (recoveries=88-107%) and good reproducible method for the analytes of interest. At relatively high concentrations, only lambda-cyhalothrin was unstable at room temperature (20-25 degrees C) and 4 degrees C over 10 days. At relatively low concentrations, acephate and acetamiprid were also unstable regardless of temperature. After 10 days storage at room temperature, 30-40% degradation of lambda-cyhalothrin was observed. It is recommended that acephate, acetamiprid, and lambda-cyhalothrin be stored at -20 degrees C or analyzed immediately after extraction.

Vitellogenin induction has been widely used as a biomarker of endocrine disruption in wildlife, but few studies have investigated its use in wild reptiles living in contaminated habitats. This study examined vitellogenin induction in Morelet's crocodiles (Crocodylus moreletii) from wetlands in northern Belize contaminated with organochlorine (OC) pesticides. Vitellogenin was measured in 381 crocodile plasma samples using a vitellogenin ELISA previously developed for this species. Vitellogenin was detected in nine samples, all from adult females sampled during the breeding season. Males and juvenile females did not contain detectable levels of vitellogenin; however, many of these animals contained OC pesticides in their caudal scutes, confirming contaminant exposure. The lack of a vitellogenic response in these animals may be attributable to several factors related to the timing and magnitude of exposure to endocrine-disrupting chemicals and should not be interpreted as an absence of other contaminant-induced biological responses.

We evaluated the use of the gas exchange rate as an ecologically relevant indicator of chemical stress in avian embryos/eggs. Northern bobwhite quail (Colinus virginianus) were exposed to octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) via feed containing nominal concentrations of 0, 12.5, 50.0, and 125.0 mg kg(-1). Metabolic rates (oxygen consumption) of developing quail eggs were then measured via respirometry to examine potential effects of HMX exposure. Metabolic rates were examined on 5, 9, and 21 d of incubation. Next, concentrations of HMX in embryos/eggs were determined by liquid chromatography-mass spectrometry. Mean (+/-SE) concentrations of HMX in eggs were 21.0+/-5.9, 1113+/-79.0, 3864+/-154.0, and 7426+/-301.1 ng g(-1) in control, low, medium and high dose groups, respectively. There were significant differences in oxygen consumption among the three embryo ages, however differences among the ages were not consistent among dose groups (age x dose group interaction p<0.0001). Oxygen consumption rates did not vary as a function of HMX in embryos (p=0.18). No evidence was observed for alterations of in ovo metabolic rates associated with HMX exposure.

Here we review mechanisms and factors influencing contaminant exposure among terrestrial vertebrate wildlife. There exists a complex mixture of biotic and abiotic factors that dictate potential for contaminant exposure among terrestrial and semi-terrestrial vertebrates. Chemical fate and transport in the environment determine contaminant bioaccessibility. Species-specific natural history characteristics and behavioral traits then play significant roles in the likelihood that exposure pathways, from source to receptor, are complete. Detailed knowledge of natural history traits of receptors considered in conjunction with the knowledge of contaminant behavior and distribution on a site are critical when assessing and quantifying exposure. We review limitations in our understanding of elements of exposure and the unique aspects of exposure associated with terrestrial and semi-terrestrial taxa. We provide insight on taxa-specific traits that contribute, or limit exposure to, transport phenomenon that influence exposure throughout terrestrial systems, novel contaminants, bioavailability, exposure data analysis, and uncertainty associated with exposure in wildlife risk assessments. Lastly, we identify areas related to exposure among terrestrial and semi-terrestrial organisms that warrant additional research.

We describe a case history of larval Hannemania sp. (Acari: Trombiculidae) infestations in two spadefoot toads (Spea spp. Anura: Pelobatidae) captured from playa wetlands in the Southern High Plains, Texas. Hannemania sp. larvae were superficially attached to the skin, not encysted, and dermal changes were not observed. Gross field examinations revealed a presumed occurrence of Hannemania sp. in Spea spp. at four of 24 wetlands in 2003 and 2004. Although other species of amphibians were present (Ambystoma tigrinum mavortium, Bufo cognatus, Bufo debilis insidior, Pseudacris clarkii, Gastrophryne olivacea, Rana blairi, Scaphiopus couchii), investigators did not observe infestations in these species. Future research should focus on identifying this Hannemania to species level and determining if it influences Spea spp. survival.

Adult Northern bobwhite quail (Colinus virginianus) were exposed via food to octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX), an energetic compound found in soils at military training installations. Depuration of HMX into eggs was examined in an initial study, and effects on egg production, hatching, growth, development, and survival of chicks were examined in a follow-up study. HMX was readily and rapidly transferred from female quail into eggs. Marked weight loss was observed in quail exposed to 125 and 250 mg/kg HMX in food, likely due to reductions in food intake rather than a toxic mechanism. In the second study, significant alterations in body mass occurred among quail at concentrations >52.5 +/- 9.3 mg/kg but not at 12.3 +/- 1.1 mg/kg in food. Treatment-related reductions in food consumption and decreases in egg laying rates were observed. No HMX-related effects were found in chick growth or survival. Quail inhabiting HMX-contaminated sites could possibly be exposed to HMX and therefore deposition of HMX into eggs is also possible. However, results of these studies further suggest that the potential for reproductive toxicity of HMX to birds is low.

Methylmercury cation (MeHg) and divalent mercury (Hg++) accumulation in liver, kidney, and brain were quantified in prairie voles (Microtus ochrogaster) at 0, 3, 6, and 12 weeks during chronic exposure to aqueous MeHg. Dose groups received deionized water or aqueous solutions containing 9, 103, or 920 ng MeHg/ml. Our study presents temporal patterns of Hg++ and MeHg concentrations in organ tissues and makes inter-tissue comparisons at each time point to illustrate the accumulation and distribution of Hg species during the study. MeHg was accumulated in tissues for 3 weeks and then concentrations plateaued. Mercury accumulated in brain, liver, and kidney to average concentrations of 510 ng/g, 180 ng/g, and 3400 ng/g, respectively. MeHg and Hg++ concentrations were roughly equivalent in liver, kidney, and urine. MeHg concentrations in brain tissue were 2 to 20 times the concentrations of Hg++. Regression analysis was also used to demonstrate the utility of urinalysis as an indicator of Hg++ and MeHg concentrations in organ tissue (p < 0.001).

An efficient sample extraction and cleanup method was developed for determination of octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) in eggs. The procedure included solvent extraction of HMX from eggs followed by cleanup using florisil and styrene-divinylbenzene (SDB) cartridges. Homogenized egg aliquots were thoroughly mixed with 10mL acetonitrile and extracted with ultrasonication for 1h. Each sample was centrifuged and all liquid was collected for cleanup. After concentration by N(2) evaporation, each extract was cleaned by florisil and SDB cartridges to remove endogenous interfering compounds. Finally, each extract was filtered through a 0.2microm PTFE membrane and stored for liquid chromatographic-mass spectrometric (LC-MS) analysis. Chromatographic separation was achieved on a reverse phase (RP) C18 column, with a mobile phase containing 60% methanol+40% 1.0mM acetic acid aqueous solution. Acetic acid was employed as mobile phase additive to form negatively charged adduct ions [M+CH(3)COO](-), and m/z=355 was quantified by selective ion monitoring (SIM). Overall recoveries from eggs containing 10, 50, 250 and 1000ng/g of HMX were 84.0%, 88.0%, 90.6% and 87.4%. A method detection limit (MDL) of 0.15ng/g was achieved.

Despite high animal diversity in the Neotropics and the largely unregulated use and disposal of pesticides and industrial chemicals in Central America, few data exist regarding accumulation of environmental contaminants in Central American wildlife. In this study we examined accumulation of metals and organochlorine (OC) pesticides in caudal scutes of crocodiles from Belize and Costa Rica. Scutes from Morelet's crocodiles (Crocodylus moreletii) from two sites in northern Belize were analyzed for metals, and scutes from American crocodiles (C. acutus) from one site in Costa Rica were analyzed for metals and OC pesticides. All scutes (n=25; one scute from each of 25 individuals) contained multiple contaminants. Mercury was the predominant metal detected, occurring in all scutes examined from both species. Other metals detected include cadmium, copper, lead, and zinc. American crocodile scutes from Costa Rica contained multiple OC pesticides, including endrin, methoxychlor, p,p'-DDE, and p,p'-DDT, all of which occurred in 100% of scutes analyzed (n=6). Mean metal and OC concentrations varied in relation to those previously reported in crocodilian scutes from other localities in North, Central, and South America. OC concentrations in American crocodile scutes were generally higher than those previously reported for other Costa Rican wildlife. Currently, caudal scutes may serve as general, non-lethal indicators of contaminant accumulation in crocodilians and their areas of occurrence. However, a better understanding of the relationships between pollutant concentrations in scutes, internal tissues, and environmental matrices at sample collection sites are needed to improve the utility of scutes in future ecotoxicological investigations.

Seven complete clutches of Morelet's crocodile (Crocodylus moreletii) eggs were collected in northern Belize and examined for organochlorine (OC) pesticide residues. The primary OC detected, p,p-DDE, was found in every egg analyzed (n=175). Other OCs detected included p,p-DDT, p,p-DDD, methoxychlor, aldrin, and endosulfan I. Concentrations of individual OCs ranged from 4 ppb (ng chemical/g egg wet weight) to greater than 500 ppb. A statistical evaluation of p,p-DDE levels in three complete clutches was used to derive the minimum number of eggs needed from a clutch to precisely determine the mean p,p-DDE concentration representative of that clutch. Sample sizes of 8 (80% confidence level) and 11 (90% confidence level) were determined to yield an accurate estimate of contaminant levels in a full clutch of eggs. The statistically recommended sample size of 11 eggs (at 90% confidence level) was successfully tested on the four additional clutches.

The Southern High Plains (SHP) is a semi-arid region in which playa wetlands are the focal points of biodiversity. Playas are highly influenced by surrounding land use. Most of the SHP is in agricultural production (primarily cotton) with a history of arsenic-containing herbicide use. Metals influence reproduction and development in amphibians. We analyzed metal residues in playa sediment and whole body tissue of Spea spp. and Bufo cognatus metamorphs from two land uses: cropland and native grassland. Cd and Ni concentrations in B. cognatus tissues differed between land uses. Metal concentrations in Spea spp. tissues did not differ between land uses. Ba was higher in Spea spp. than B. cognatus collected from the same grassland playas, indicating differential habitat use. No correlations between sediment and tissue concentrations were found. Land use appeared to have little influence on metal concentrations and levels were below those known to cause effects in amphibians.

The purpose of this study was to develop an immunoassay for vitellogenin in Morelet's crocodile (Crocodylus moreletii). Blood was collected from wild-caught crocodiles in Belize. Plasma samples from adult females taken during the breeding season were used for vitellogenin purification and samples from adult males were used for comparison. No differences were detected between males and females for plasma total protein concentration, as measured by Coomassie assay. However, denaturing polyacrylamide gel electrophoresis (SDS-PAGE) revealed that female plasma contained a 210-kDa protein, presumably the vitellogenin monomer, that was absent in adult male plasma. The identity of the putative vitellogenin was confirmed by its cross-reactivity in Western blots with a vitellogenin antiserum that was generated against a conserved vitellogenin peptide sequence. Crocodile vitellogenin was purified by two successive rounds of DEAE chromatography. The purified protein had an apparent molecular mass of 450 kDa, as determined by gel filtration chromatography, and 210 kDa on SDS-PAGE. An indirect enzyme-linked immunosorbent assay (ELISA) was then developed for C. moreletii vitellogenin. The detection limit of the assay was 20.0 ng/mL. The intra- and inter-assay coefficients of variation were 5.3% and 9.8%, respectively. The recovery of vitellogenin diluted into male plasma was 94.7%. The ELISA assay revealed that vitellogenin levels of adult female plasma during the breeding season ranged from 1.8 to 3.1 mg/mL with a mean of 2.5+/-0.25 mg/mL. No vitellogenin was detected in adult male plasma. Induction of vitellogenin in Morelet's crocodile may be a useful model system for field studies of crocodile reproduction and for investigations of endocrine disruption in this species.

We live-trapped 40 northern pocket gophers across two years from the Anaconda Smelter Superfund Site, Anaconda, Montana, USA, to determine their exposure to five metal contaminants and effects of exposure on selected measurements. Soil, gopher blood, liver, kidney, and carcass samples were analyzed for arsenic, cadmium, lead, copper, and zinc. Hematological parameters, kidney and liver porphyrins, and red blood cell delta-aminolevulinic acid dehydratase (ALAD) activity were also measured. Micronutrients Cu and Zn were detected in all tissues analyzed, and Cd, Pb, and As were detected less frequently. We report differences in metal distribution among different tissues and differences in bioaccumulation for different metals within the same tissue. No significant differences were observed in concentrations of Zn or Cu in any tissue across the study site, but relationships between lead in soil and lead in carcass proved especially strong (r2 = 0.80; p < 0.001; n = 18). Among biomarker data, we observed a negative relationship between concentration of lead in the soil and ALAD activity in gophers with detectable concentrations of lead in their blood (r2 = 0.45; p = 0.006; n = 15). Results of this study suggest that northern pocket gophers are useful biomonitors of environmental Pb, Cd, and As contamination, and their broad geographic range across North America could allow them to be an important component of site-specific metals assessments.

Despite their ecological importance and global decline, snakes remain poorly studied in ecotoxicology. In this study, we examined organochlorine (OC) pesticide and mercury accumulation in cottonmouths (Agkistrodon piscivorus) living on a contaminated site in northeastern Texas (USA). Mercury and p,p'-dichlorodiphenyldichloroethylene (p,p'-DDE) were detected in all snakes examined. Other OCs, including p,p'-dichlorodiphenyltrichloroethane (p,p'-DDT), methoxychlor, aldrin, and heptachlor, also were detected, but less frequently. Concentrations of p,p'-DDE were higher in fat than in liver, while mercury concentrations were highest in liver, followed by kidney and tail clips. One animal contained the highest mercury concentration yet reported for a snake (8,610 ng/g). Mercury concentrations in liver and kidney were higher in males than females, while no intersex differences in p,p'-DDE concentrations were observed. Concentrations of p,p'-DDE in fat were correlated positively with body size in male cottonmouths but not females, suggesting a slower rate of accumulation in females. Body size strongly predicted mercury concentrations in liver, kidney, and tail clips of both sexes. Tail clips were strong predictors of mercury in liver and kidney in males but not females, suggesting possible sex-dependent differences in mercury toxicokinetics. Both long-term field studies and controlled laboratory investigations are needed to adequately assess the response of cottonmouths to chronic contaminant exposure.

Perchlorate is a water soluble anion that is readily accumulated in vegetation. It inhibits uptake of iodide into thyroid gland tissue, thereby reducing production of thyroid hormones. Potential raccoon food items including berries, fish, and vegetation collected at a contaminated site contained quantifiable concentrations of perchlorate as determined by ion chromatography. Therefore, we monitored resident raccoons for exposure to perchlorate by examining plasma perchlorate and thyroid hormone concentrations. Resulting analytical data failed to demonstrate perchlorate exposure among raccoons that likely consumed food items collected along perchlorate-contaminated water bodies. There were no correlations between triiodothyronine or thyroxine and thyroid stimulating hormone concentrations, but triiodothyronine concentrations in raccoon plasma were significantly higher in 2000 than in 2001 (p = 0.0081). These data suggest that natural attenuation and remedial efforts initiated in January of 2001 may have reduced perchlorate exposure among raccoons inhabiting this site from 2000 to 2001. Temporal, spatial, and analytical factors limited our ability to quantify exposure among raccoons, however, our data do not indicate that raccoons currently inhabiting this site are at risk for significant exposure to perchlorate and subsequent effects.

Water, soil, vegetation, and rodents were collected from three areas along the Las Vegas Wash, a watershed heavily contaminated with perchlorate. Perchlorate was detected at elevated concentrations in water, soil, and vegetation, but was not frequently detected in rodent liver or kidney tissues. Broadleaf weeds contained the highest concentrations of perchlorate among all plant types examined. Perchlorate in rodent tissues and vegetation was correlated with perchlorate concentrations in soil as expected, however rodent residues were not highly correlated with plant perchlorate concentrations. This indicates that soil may be a greater source, or a more constant source of perchlorate exposure in rodents than vegetation.

Passerine eggs make useful biomonitors of environmental pollutants. Among passerines, it is not known whether organochlorine contaminants in eggs within the same clutch are independent observations or follow a laying order effect. Intraclutch variation of DDE (1,1-dichloro-2,2-bis[(p-chlorophenyl)]ethylene) concentrations was studied in eggs collected from prothonotary warblers (Protonotaria citrea) and European starlings (Sturnus vulgaris) nesting on National Priority List sites in lower Alabama and central Colorado, respectively. All 209 eggs collected for this study contained detectable levels of DDE. Mean concentration of DDE across all prothonotary warbler eggs (mean 8.71 microg/g +/- 1.19, n = 20) was almost two orders of magnitude greater than mean concentrations of DDE in all starling eggs (mean 0.70 microg/g +/- 0.06, n = 189). In both species, there was a large amount of variability among individual eggs of the same clutch and no significant relationship between laying order and DDE concentration. Variation among eggs laid in the same sequential order was high and effectively masked any potential trends in laying order effect. We hypothesized that the variability was caused by the spatial heterogeneity of DDE on our study sites, the nature of egg development within a female passerine, or a combination of these factors. Investigators focusing on lipophilic contaminants should exercise caution when making inferences about contaminant concentrations in an entire clutch of passerine eggs after the collection and analysis of a single egg because our data show that DDE levels in a single egg collected for analysis do not consistently reflect DDE levels in the eggs remaining in the nest., (Copyright 2004 Springer Science + Business Media, Inc.)

Recent studies examined the utility of the chorioallantoic membrane (CAM) as a nonlethal, noninvasive indicator of environmental contaminant exposure in oviparous wildlife. The CAM is a highly vascularized extraembryonic membrane that functions as a site for respiration, nutrient transport, and waste storage during embryonic development. After hatching, the CAM is usually discarded with the eggshell and can be used for chemical residue analysis. Chorioallantoic membranes have been used successfully to examine contaminant exposure and predict chemical concentrations in multiple species of birds and reptiles. In this study, we examined organochlorine (OC) pesticide concentrations in CAMs from eggs of Morelet's crocodiles (Crocodylus moreletii) from northern Belize. Multiple OCs were detected in crocodile CAMs, including aldrin, dieldrin, endrin, dichlorodiphenyltrichloroethane, dichlorodiphenyldichloroethane, dichlorodiphenyldichloroethylene (DDE), heptachlor, lindane, and methoxychlor. Number and concentrations of OC compounds in CAMs were variable. The most prevalent contaminant detected was DDE, which occurred in 69% of CAMs, with concentrations ranging from 0.3 parts per billion (ppb) to 17.0 ppb. The OC burdens in crocodile CAMs confirm contamination of eggs and suggest exposure in embryos and maternal females. These results further support the use of CAMs as qualitative indicators of OC exposure in oviparous wildlife. The efficacy of this sampling technique in the field will depend on the logistics and cost associated with CAM collection and the specific life history traits of the wildlife species.

Perchlorate exposure and potential effects were evaluated in large mammals by monitoring heifer calves placed on a site with access to streamwater fed by a perchlorate-contaminated groundwater spring ( approximately 25 ng/mL). Blood was collected from the two calves on the site (and two control calves from an uncontaminated site) approximately every 2 weeks for analysis of perchlorate residues and thyroid hormones. During the 14 week study, perchlorate was detected (detection limit = 13.7 ng/mL) in blood plasma twice (15 ng/mL and 22 ng/mL) in one of the heifer calves drinking perchlorate-contaminated water on consecutive sampling periods 4 and 6 weeks after the beginning of perchlorate exposure. Constant exposure to 25 ppb perchlorate in drinking water had no effect on circulating thyroid hormones (T(3) and T(4)) in the heifer calves.

An investigation involving raccoons (Procyon lotor) as a sentinel species at the Paducah Gaseous Diffusion Plant (PGDP) in Western Kentucky (USA) delineated the extent of exposure to polychlorinated biphenyls (PCBs) and PCB spatial distribution. Raccoon exposure to PCBs was demonstrated through analysis of subcutaneous fat, abdominal fat, liver, and brain tissues from raccoons collected at the PGDP but also was clearly evident in raccoons from a reference area situated along the Ohio River (USA). Raccoons with the highest tissue PCB concentrations appeared to be those inhabiting areas nearest the plant itself and most likely those that ventured into the plants interior. Male raccoons at the PGDP had similar concentrations of total PCBs in subcutaneous fat (1.86 +/- 0.64 microg/g) as males from the reference site (1.41 +/- 0.35 microg/g), but females had higher PCB body burdens than those at the reference site (9.90 +/- 6.13 microg/g vs 0.75 +/- 0.40 microg/g). Gross measurements of exposure to radiation-producing materials revealed that counts per minute exceeded background in 61% of PGDP raccoons compared with 27% at the reference site and five raccoons at the PGDP had beta counts that were more than twice the background. Differences among trapping success, growth rates, and serum chemistry parameters were noted but may have been related to habitat and other environmental and population density factors.

Mercury occurrence in prothonotary warblers (Protonotaria citrea) was evaluated over two years in southern Alabama. Mercury was found in warbler nestlings and adults inhabiting National Priority List (NPL) sites in McIntosh, Alabama. Food items that were collected from nestlings also contained elevated mercury. When mercury concentrations in soil, food, and nestling were plotted at each nest box location, the distribution of mercury in the three matrices yielded information that direct bioaccumulation factors could not. There were site differences in mercury accumulation in nestlings inhabiting the NPL sites. Nestling mercury accumulation correlated with solid mercury concentrations near the nest box where the nestling was raised. Trophic transport of mercury was poorly defined by mercury in food; however, closer examination of prey items shows that food source influences accumulation. Mercury distributions in matrices provide useful information of uptake that can be integrated with risk assessment endpoints.