Radiographic determination of fecundity: Is the technique safe for developing turtle embryos?
No abstract available.
Chelonian Conservation and Biology↗
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Unusual subterranean aggregations of the California Giant Salamander, Dicamptodon ensatus
Larval Dicamptodon are one of the most abundant vertebrates in headwater streams in the Pacific Northwest. Their numbers and biomass can exceed those of all other amphibians, and of salmonid fishes. By contrast, metamorphosed Dicamptodon are only found infrequently, usually during formal surveys using pitfall traps, cover boards, or time constrained surveys However, we found two aggregations (23 and 27 individuals) of metamorphosed Dicamptodon ensatus during a culvert removal project at Point Reyes National Seashore, California. Furthermore, we found an additional 23 terrestrial D. ensatus in terrestrial habitat adjacent to the culverts. We did not expect these aggregations because metamorphosed individuals are so rarely encountered, and aggregations are likely to increase competition and predation in a species known to feed regularly on vertebrate prey. Deteriorating culverts might provide an unusually high-quality habitat that leads to aggregations such as we describe. Our observations may provide insight into the natural haunts of D. ensatus—underground burrows or caverns—and if so, then aggregations may be normal, but rarely seen.
Herpetological Conservation and Biology↗
Frogs on the beach: Ecology of California Red-legged Frogs ( Rana draytonii ) in coastal dune drainages
California Red-legged Frogs (Rana draytonii) are typically regarded as inhabitants of permanent ponds, marshes, and slow-moving streams, but their ecology in other habitats, such as drainages among coastal dunes, remains obscure. Because coastal dune ecosystems have been degraded by development, off-highway vehicle use, stabilization, and invasive species, these unique ecosystems are the focus of restoration efforts. To better understand the ecology of California Red-legged Frogs in coastal dune ecosystems and to avoid and minimize potential negative effects of dune restoration activities on these rare frogs, we studied their spatial ecology, habitat selection, and survival in coastal dune drainages at Point Reyes National Seashore, California, USA. All 22 radio-marked frogs remained in their home drainages throughout the spring and summer of 2015 and, with some notable exceptions, most remained close to water. Local convex hull home ranges of four out of five California Red-legged Frogs with > 20 observations in dunes were < 1,600 m2 . At the population level, frogs were 1.7 (95% credible interval, 1.2‒4.4) times more likely to select sites 1 m closer to water, and were 83 (2.0‒17,000) times more likely to select sites with 10% greater percentage cover of logs that served as refuges from environmental extremes and predators. On average, California Red-legged Frogs avoided the invasive plants Iceplant (Carpobrotus edulis) and European Beachgrass (Ammophila arenaria). Frogs were 0.68 (0.32‒0.89) and 0.55 (0.24‒0.75) times as likely to select areas that had 10% greater cover of these plants, respectively. Assuming constant risk of mortality, California Redlegged Frogs had an annual survival rate of 0.70 (0.27‒0.96) in coastal dune drainages. Our results indicate that coastal dune drainages provide a locally important habitat for California Red-legged Frogs. Restoration practices that maintain wetted drainages with logjams are likely to benefit California Red-legged Frogs.
Herpetological Conservation and Biology↗
Amphibians, pesticides, and the amphibian chytrid fungus in restored wetlands in agricultural landscapes
Information on interactions between pesticide exposure and disease prevalence in amphibian populations is limited, especially from field data. Exposure to certain herbicides and insecticides has the potential to decrease the immune response in frogs, which can potentially lead to increased abundance of Batrachochytrium dendrobatidis (Bd) zoospores on individuals and in the wetlands. In contrast, exposure to certain fungicides can decrease Bd abundance on frog skin. We examined the relationships between the abundance of Bd on the skin of individual Boreal Chorus Frogs (Pseudacris maculata) and the concentrations of pesticides in the water and in frog tissue at six agriculturally dominated wetlands in Iowa, USA. We collected frogs from each wetland, swabbed them for Bd, and analyzed their tissues for a suite of fungicides, herbicides, and insecticides. We collected surface water from the wetlands and we analyzed it for the same suite of pesticides. We observed no relationship between Bd zoospores on the skin of individual frogs and the concentrations of total pesticides, total herbicides/insecticides and total fungicides in frog tissue. Similarly, we observed no relationship between Bd zoospore abundance in water and the concentration of total pesticides or total herbicides in water. However, we observed a negative relationship between Bd zoospore abundance in water and neonicotinoid concentrations in surface water. Negative results are seldom reported but can be important contributors to a more complete understanding of the complex and potentially synergistic relationships between disease and pesticides. Data from field studies on these relationships are particularly scarce. As our laboratory understanding of these relationships expands, the need for field based, or applied, studies grow.
Iowa↗
Spatiotemporal ecology of Apalone spinifera in a large, Great Plains river ecosystem
Sparse information exists about the ecology of Spiny Softshell Turtles (Apalone spinifera) in large rivers, at the northwestern extent of their natural range, and in Montana, where they are disjunct from downstream populations and a State Species of Concern. We determined spatiotemporal ecology of 47 female and 12 male turtles from 2009 through 2012 and identified fundamental habitats in the Missouri River in east-central Montana. Movement rates of females were greater than those of males and peaked before nesting. Movement rates of males peaked before overwintering, and movement rates of both sexes were minimal in winter. Home range sizes were not different between sexes, varied among individuals and seasons, and were similar to those reported elsewhere in their northern range. Turtles aggregated and showed interannual fidelity to separate and disparate habitats in different seasons. Turtles often chose fine substrates, tributary confluences, and reaches with islands during summer and mainstem outside bends in the winter. They inhabited shallow, slow water velocity areas from May to September. They inhabited deeper, moderate velocity areas from October to April. We did not observe ice jams and associated riverbed scour at hibernacula, but did observe them elsewhere. Ice jams may be spatially predictable and influence the distribution of riverine turtles during autumn and winter. Preservation of dissimilar habitats used during major portions of the life cycle (lateral habitats, islands, and hibernacula) and natural streamflow patterns, which influenced timing of habitat availability and turtle movement, may facilitate continued existence of Spiny Softshell Turtles in the Missouri River in Montana
Herpetological Conservation and Biology↗
Estimating Mudpuppy (Necturus maculosus) abundance in the Lamoille River, Vermont, USA
The Mudpuppy (Necturus maculosus) is classified as a Species of Greatest Conservation Need by the state of Vermont. There is concern regarding status of populations in the Lake Champlain basin because of habitat alteration and potential effects of 3-trifluromethyl-4-nitrophenol (TFM), a chemical used to control Sea Lamprey (Petromyzon marinus). The purpose of our research was to assess Mudpuppy capture methods and abundance in the Lamoille River, Vermont, USA. We sampled Mudpuppies under a mark-recapture framework, using modified, baited minnow traps set during two winter-spring periods. We marked each Mudpuppy with a passive integrated transponder (PIT) tag and released individuals after collecting morphological measurements. We collected 80 individuals during 2,581 trap days in 2008–2009 (year 1), and 81 individuals during 3,072 trap days in 2009–2010 (year 2). We estimated abundance from spring trapping periods in 2009 and 2010, during which capture rates were sufficient for analysis. Capture probability was low (< 0.04), but highest following precipitation events in spring, during periods of higher river flow, when water temperatures were approximately 3 to 6° C. During October 2009, management agencies treated the Lamoille River with TFM. Surveyors recovered more than 500 dead Mudpuppies during the post-treatment assessment. Overall, Mudpuppy captures did not change between sampling periods; however, we captured fewer females during year 2 compared to year 1, and the sex ratio changed from 0.79:1 (M:F) during year 1 to 3:1 (M:F) during year 2. Our data may help wildlife managers assess population status of Mudpuppies in conjunction with fisheries management techniques.
Vermont↗
Using passive integrated transponder (PIT) systems for terrestrial detection of blue-spotted salamanders (Ambystoma laterale) in situ
Pure-diploid Blue-spotted Salamanders ( Ambystoma laterale ) are the smallest members of the family Ambystomatidae which makes tracking with radio-transmitters difficult because of small battery capacity. Passive integrated transponder (PIT) tags provide another tracking approach for small fossorial animals such as salamanders. We evaluated the use of portable PIT tag readers (PIT packs) to detect PIT tag-implanted pure-diploid Blue-spotted Salamanders in situ. We also examined the detection probability of salamanders with PIT tags held in enclosures in wetland and terrestrial habitats, as well as the underground detection range of PIT packs by scanning for buried tags not implanted into salamanders. Of the 532 PIT tagged salamanders, we detected 6.84% at least once during scanning surveys. We scanned systematically within a 13.37 ha area surrounding a salamander breeding pool on 34 occasions (~119 hours of survey time) and detected PIT tags 74 times. We detected 55% of PITs in tagged salamanders and 45%were expelled tags. We were able to reliably detect buried PIT tags from 1–22cm below the ground surface. Because nearly half the locations represented expelled tags, our data suggest this technique is inappropriate for future studies of pure-diploid Blue-spotted Salamanders, although it may be suitable for polyploid Blue-spotted Salamanders and other ambystomatid species, which are larger in size and may exhibit higher tag retention rates. It may also be prudent to conduct long-term tag retention studies in captivity before tagging and releasing salamanders for in situ study, and to double-mark individuals.
Herpetological Conservation and Biology↗
Prey preferences and prey acceptance in juvenile Brown Treesnakes (Boiga irregularis)
On the Pacific island of Guam, control of the invasive Brown Treesnake (Boiga irregularis) relies largely on methods that use mice as bait. Juvenile B. irregularis feed primarily on lizards and their eggs, but little is known about their prey preference. We conducted an experiment to investigate preferences for, and acceptance of, dead geckos, skinks, and neonatal mice, in juvenile B. irregularis ranging from 290 mm to ca. 700 mm snout-vent length (SVL). Snakes of all sizes showed a preference for geckos over skinks and neonatal mice. Geckos were the first prey chosen in 87% of 224 initial trials (56 snakes subjected to four trials each; 33% would be expected from a random choice). The smallest snakes had the most pronounced preference. Although many of the snakes accepted neonatal mice and/or skinks, some snakes of all sizes were reluctant to feed on anything but geckos, especially when well fed. We also addressed the hypothesis that repeated encounters with a particular prey type increase a snake's preference for that prey. Our study does not support this hypothesis. Our results suggest that control methods relying solely on rodent bait may be inefficient for targeting snakes < 700 mm SVL and that individual heterogeneity in prey preference may cause a significant part of this juvenile cohort to be completely refractory to capture with rodent bait, even if the bait is dead and small enough to be readily swallowed.
Herpetological Conservation and Biology↗
Mitigating road mortality of diamond-backed terrapins (Malaclemy's terrapin) with hybrid barriers at crossing hot spots
Roads represent a pervasive feature on most landscapes that can pose multiple threats to wildlife populations and substantial challenges for management. To be effective, management strategies must often target where threats are most concentrated. Road mortality and nest predation are well-documented threats to Diamond-backed Terrapins ( Malaclemys terrapin ) across the majority of their range, including the 8.7-km causeway to Jekyll Island, Georgia, USA, where both are predicted to contribute to population declines if left unmitigated. From 2009 to 2014, we used intensive road surveying to identify spatial peaks (hot spots) of terrapin crossing activity and road mortality and exploit these as targets for management. In 2011, we deployed a hybrid barrier composed of nest boxes, which were designed to prevent terrapins from accessing the road and mitigate nest predation, at one hot spot while leaving two other hot spots unmanaged. We evaluated the impact of the barrier on terrapin emergences on the causeway under a Before-After-Control-Impact (BACI) design, and a companion study evaluated the effects of nest boxes on nest predation rates. We estimated a 57% reduction in annual terrapin emergences at the barrier site compared to no measurable change at control hot spots. Our findings support the use of hybrid barriers for simultaneously addressing road mortality and nest predation for other terrapin populations at risk to these threats. Our approach highlights the need to design feasible but robust management strategies that target spatial peaks of road mortality while addressing additional threats contributing to population declines of terrapins and other species.
Georgia↗
Habitat selection, movement patterns, and hazards encountered by northern leopard frogs (Lithobates pipiens) in an agricultural landscape
Telemetry data for 59 Northern Leopard Frogs (Lithobates pipiens) breeding in ponds in Houston and Winona Counties, MN; 2001-2002. Agricultural intensification is causing declines in many wildlife species, including Northern Leopard Frogs (Lithobates pipiens). Specific information about frog movements, habitat selection, and sources of mortality can be used to inform conservation-focused land management and acquisition. We studied Northern Leopard Frogs in southeastern Minnesota, part of the Driftless Area ecoregion, characterized by hills and valleys and a mix of agriculture, forests, small towns and farmsteads. In this area, small farm ponds, originally built to control soil erosion are used by the species for breeding and wintering in addition to riparian wetlands. But, this agricultural landscape may be hazardous for frogs moving between breeding, feeding, and wintering habitats. We surgically implanted transmitters into the peritoneal cavity of 59 Northern Leopard Frogs and tracked them from May to October 2001-2002. The total distance traveled by radio-tagged frogs ranged from 12 to 3316 m, the 95% home range averaged 5.3 ± 1.2 (SE) ha, and the 50% core area averaged 1.05 ± 0.3 (SE) ha. As expected, Northern Leopard Frogs selected wetlands over all other land cover classes and row crops were generally avoided at all levels of selection. Only a few tracked frogs were successful at dispersing (n = 6). Most frogs attempting to disperse (n =31) ended up missing (n = 14), died due to mowing (n = 8), or were recorded as transmitter failure (n = 2) or unknown mortalities (n = 1). For the conservation of Northern Leopard Frogs in this agricultural setting, we must consider both the aquatic and the terrestrial needs of this species. Conservation agencies that restore, manage, and acquire wetlands should consider the hazards posed by land uses adjacent to frog breeding and wintering sites and plan for movement corridors between these locations. For example, grasslands that are mowed or hayed between April and October in the north central U.S. and are adjacent to wetlands, pose a direct threat to frogs because these cultivated grasslands are primary locations for summer occupancy. When conservation land managers are selecting sites for acquisition or restoration they should avoid investments that will situate the wetland adjacent to heavily travelled roads and agricultural lands likely to be mowed or hayed. Increasing habitat amount and quality at amphibian breeding, feeding and wintering sites should reduce the energy required and hazards associated with moving long distances. Large, diverse wetlands probably provide all of the requirements needed by Northern Leopard Frogs for survival including food, shelter, breeding and overwintering areas.
Minnesota↗
Stable isotope comparison between mantle and foot tissues of two freshwater unionids: Implications for food web studies
Unionid mussels are a key taxon for stable isotope studies of aquatic food webs, often serving as the primary integrator of the pelagic baseline. Past isotope studies with mussels have commonly used either foot tissue or mantle tissue, but no study has yet to quantify the relation of both carbon and nitrogen isotopes between these two tissue sources. This makes it difficult to justify cross-study comparisons when different tissue compartments and different species were used as the basis of food web models. Therefore, we collected foot and mantle tissues from two common mussel species, Amblema plicata and Fusconaia flava , from lotic and lentic sites in the Upper Mississippi and St. Croix rivers (Minnesota/Wisconsin). Paired tissue samples from each individual were analyzed for stable isotopes of nitrogen and carbon. There were strong relations between tissue types for both isotopes between species ( r 2 > 0.93). Paired t -tests indicated that there were statistically significant differences between the tissue sources in some instances, but the difference (0.04–0.21‰) was less than the analytical precision of the mass spectrometer (circa 0.2–0.3‰). We conclude that the isotopic values from these two tissue sources are biologically comparable and recommend that researchers use the tissue source and extraction technique that minimizes stress to the mussels. We also tested for significant differences between species within a site for either isotope or tissue type and found no statistically significant difference between species with the exception of carbon in foot tissue at two sites. The highly correlated isotopic response supports the interchangeable use of both tissue compartments and both species. These findings support comparisons between studies whether the results were based on either of these tissues or the two species studied. Comparability will also simplify sampling designs, save time, and save money for processing samples without diminishing the usefulness of the data.
Minnesota, Wisconsin↗
Status of the threatened Chiricahua Leopard Frog and conservation challenges in Sonora, Mexico, with notes on other ranid frogs and non-native predators
In North America, ranid frogs (Ranidae) have experienced larger declines than any other amphibian family, particularly species native to the southwestern USA and adjacent Mexico; however, our knowledge of their conservation status and threats is limited in Mexico. We assessed the status of the federally listed as threatened (USA) Chiricahua Leopard Frog ( Lithobates chiricahuensis ) in Sonora, Mexico, based on a search of museum specimens, published records, unpublished accounts, and surveys from 2000–2016 of 84 sites within the geographical and elevational range of the species. We also provide information on occurrence of three other native ranid frog species encountered opportunistically during our surveys. The Chiricahua Leopard Frog is known in Sonora from only 20 historical (pre-2000) localities. Searches of three historical sites did not reveal any Chiricahua Leopard Frogs; however, we found it at three previously undocumented sites in 2016, all near Cananea. To our knowledge, these records are the first observations of Chiricahua Leopard Frogs in Sonora since 1998. Differences in conservation status between the USA and Sonora are likely due to differing magnitude and distribution of threats and a comparatively aggressive recovery program in the USA. For example, key non-native predators important in the decline of the Chiricahua Leopard Frog are much less widespread in Sonora compared to the southwestern USA, but there are fewer protections and recovery actions for the frog in Sonora than in the USA. Additional surveys for the Chiricahua Leopard Frog and other amphibians in Sonora should be a priority to fully assess threats and conservation status.
Sonora↗
Effects of short-term, outdoor head-starting on growth and survival in the mojave desert tortoise (gopherus agassizii)
The combination of life-history traits that makes some turtle species vulnerable to population declines also limits their ability to recover even after threats have been addressed. Because juvenile turtle survival is typically lower than adult survival, head-starting, the process of rearing juveniles through one of their most vulnerable periods, may be a useful recovery tool. We evaluated short-term, outdoor head-starting in Mojave Desert Tortoises ( Gopherus agassizii ) by comparing growth and survival among three treatments: (1) juveniles reared in outdoor predator-resistant enclosures and receiving low (LOW) or (2) high levels of rain supplementation (HIGH); and (3) free-ranging animals released 0-18 mo after hatching (FIELD). Juveniles from the HIGH treatment had higher annual growth (12.7 mm midline carapace length [MCL] per year) than juveniles from the LOW or FIELD treatments (10.7 mm). Annual growth also varied among years, presumably due to variation in rainfall. Annual survival was high (0.94 ± 0.01) for both LOW and HIGH treatments; MCL at hatching had a weak positive effect on survival probability (effect size: 0.42 ± 0.35). Annual survival of FIELD animals averaged 0.48 ± 0.09. There was no effect of size at release (40.8-61.5 mm MCL) on post-release survival of FIELD animals, suggesting that the greatest benefit of short-term outdoor head-starting is increasing survival during the head-start period. Although releasing at larger sizes (100 mm MCL) has been recommended, slow growth in tortoises would require extended outdoor head-starting periods. Indoor rearing, which has been successfully implemented with other turtle species, may increase growth rates of juvenile Desert Tortoises and warrants future study as a conservation technique.
Herpetological Conservation and Biology↗
Reproductive frequency and size-dependence of fecundity in the Giant Gartersnake (Thamnophis gigas)
How reproductive output changes with age or size is a key life-history trait that can affect which demographic rates most influence population growth. Although many studies have investigated the reproductive ecology of gartersnakes, we know little about reproduction in the threatened Giant Gartersnake, Thamnophis gigas . We used X-radiography to determine reproductive status and estimated fecundity for 73 female T. gigas collected from several regions within the range of this species in the Sacramento Valley of California, USA, and synthesize these data with data from litters born in captivity to improve our understanding of reproduction in this species. Average total litter size determined from X-rays (15.9) and captive-born litters (15.5) are within the ranges reported from other gartersnakes, but captive-born litters had high rates of stillbirth. Only 154 of 202 neonates from captive snakes were born alive, and seven of 13 litters contained at least one stillborn neonate. We found that fecundity was positively related to maternal snout-vent length, and some evidence that larger litters contained smaller neonates. The proportion of X-rayed females that were gravid was 0.50 in 2014, 0.47 in and 2015, and 0.64 in 2016. Central California experienced an exceptional drought from 2012–2015, which may have affected the reproductive output and frequency of T. gigas . Our estimates of reproductive frequency and size-dependent fecundity in T. gigas provide valuable information that can be used in demographic models of this threatened species. Our results demonstrate that X-radiography is a useful, minimally invasive means to study fecundity in wild populations of snakes.
California↗
Long-term spotlight surveys of American alligators in Mississippi, USA
Accurate population estimates and assessments of trajectory are an essential part of harvest management for game species and conservation action plans for protected species. Long-term monitoring can lead to ecological understanding by identifying biotic and abiotic drivers of population dynamics. Spotlight surveys are a widely used method to monitor abundance and size-class structure of crocodilian populations. The American Alligator (Alligator mississippiensis) has recovered from significant population reductions in the southeastern United States. The Mississippi Department of Wildlife, Fisheries, and Parks (MDWFP) has conducted alligator spotlight surveys since 1971 to monitor populations. We analyzed this long-term alligator survey dataset to assess possible trends in counts as a proxy for potential population changes. We tested for a positive trend in count data over 46 y and evaluated covariates that could influence counts to assist future survey protocols. Alligator counts during 1971– 2016 increased across survey routes in Mississippi. This observed positive response may represent an increase of the alligator population in Mississippi as a result of conservation benefits accrued from improved wetland conditions and species-specific management policies. Evaluation of survey covariates indicated recent rainfall and increasing wind velocity had negative effects on alligator counts while increasing water temperature had a positive effect. Implementing robust survey techniques will improve the reliability of alligator monitoring data and their application to the management of alligator populations. Further, these improved approaches may be useful to other conservation and management agencies as well as for other crocodilian species.
Mississippi↗
Efficacy of automatic vocalization recognition software for anuran monitoring
Surveys of vocalizations are a widely used method for monitoring anurans, but it can be difficult to coordinate standardized data collection across a large geographic area. Digital automated recording systems (ARS) offer a low-cost method for obtaining samples of anuran vocalizations, but the number of recordings can easily overwhelm human listeners. We tested Song Scope, an automatic vocalization recognition software program for personal computers to determine if this type of machine learning approach is currently a viable solution for anuran monitoring. For three species, Song Scope scanned more than 200 h of recordings in 3-20 h at the settings we chose. The software misidentified true calls (false positive) at rates of 2.7%-15.8% per species and failed to detect calls (false negative) in 45%-51% of recordings. There exists a tradeoff between false positive and false negative errors, which can be adjusted by setting the minimum criteria for the recognition software. Users of this approach should carefully consider their reasons for monitoring and how they intend to use the data before creating a large monitoring network.
Louisiana↗
Conservation genetics of imperiled striped whipsnake in Washington
Conservation of wide-ranging species is aided by population genetic information that provides insights into adaptive potential, population size, interpopulation connectivity, and even extinction risk in portions of a species range. The Striped Whipsnake (Masticophis taeniatus) occurs across 11 western U.S. states and into Mexico but has experienced population declines in parts of its range, particularly in the state of Washington. We analyzed nuclear and mitochondrial DNA extracted from 192 shed skins, 63 muscle tissue samples, and one mouth swab to assess local genetic diversity and differentiation within and between the last known whipsnake populations in Washington. We then placed that information in a regional context to better understand levels of differentiation and diversity among whipsnake populations in the northwestern portion of the range of the species. Microsatellite data analyses indicated that there was comparable genetic diversity between the two extant Washington populations, but gene flow may be somewhat limited. We found moderate to high levels of genetic differentiation among states across all markers, including five microsatellites, two nuclear genes, and two mitochondrial genes. Pairwise state-level comparisons and dendrograms suggested that Washington whipsnakes are most closely related to those in Oregon, and distinct from Idaho, Nevada, and Utah, approximately following an isolation by distance model. We conclude that Washington populations of whipsnakes have experienced recent isolating events, but they have yet to lose genetic diversity. The longevity and high vagility of the species may provide opportunity for conservation of whipsnakes in the state as long as shrubland habitat is available
Washington↗
Environmental contamination and unusual snake mortality in an urban national wildlife refuge
The National Wildlife Refuge (NWR) System protects ~150 million acres of land and water in the United States and provides habitat for >2,000 native vertebrates species. Although legally protected, wildlife populations within these refuges can be threatened by anthropogenic activities. The lack of knowledge about such threats has the potential to undermine biodiversity conservation. We investigated patterns of wildlife mortality among three NWRs in the northern U.S. Gulf Coast. Visual surveys (958 total search-hours) were conducted for live or dead amphibians and reptiles at Big Branch, Bogue Chitto, and Bayou Sauvage NWRs. An exceptionally high incidence of snake mortality (>800% above background, P < 0.001) was observed at one site (Haul Road, Bayou Sauvage NWR). Mortality affected seven species and recurred for 29 months. Intact carcasses were subject to histopathological examination and diagnostic testing. Cause of death was undetermined (n = 9) or attributed to various pathogens (n = 5). To investigate possible underlying causes of this unusual mortality, water exposure challenges and soil toxicological analyses (n = 6 and 11 sites, respectively) were conducted on samples from Bayou Sauvage NWR, given the known potential for anthropogenic contamination in the refuge. In a controlled experiment, survival of Anaxyrus fowlerii embryos exposed to water from Recovery Road (i.e., 250m from a landfill) and Haul Road was reduced 92% and 48%, respectively, compared to four reference sites in the refuge. Sediment analyses suggested at least seven distinct sources of contaminants in Bayou Sauvage NWR. Potential sources included a landfill, illegal dumping, vehicle emissions, pesticide migration, and an active hydrocarbon pipeline leak discovered 600 m from Haul Road. Synthesis and applications: Although further research is needed to determine whether the observed mortality and contamination are related, these collective findings suggest that anthropogenic factors may threaten the ecological integrity of one of the United States largest urban wildlife refuges. More broadly, our study highlights the critical need for species inventories, baseline data, and systematic monitoring in NWRs. Such information is essential to detecting and mitigating anthropogenic threats to biodiversity conservation in protected areas.
Louisiana↗