Using genome-wide data to ascertain taxonomic status and assess population genetic structure for Houston toads (Bufo [= Anaxyrus] houstonensis).

The Houston toad (Bufo [= Anaxyrus] houstonensis) is an endangered amphibian with a small geographic range. Land-use changes have primarily driven decline in B. houstonensis with population supplementation predominant among efforts to reduce its current extinction risk. However, there has been historic uncertainty regarding the evolutionary and conservation significance of B. houstonensis. To this end, we used 1170 genome-wide nuclear DNA markers to examine phylogenetic relationships between our focal taxon, representatives of the Nearctic B. americanus group, and B. nebulifer, a sympatric Middle American species. Phylogenetic analyses indicate B. houstonensis is a taxon that is distinct from B. americanus. We corroborated such genetic distinctiveness with an admixture analysis that provided support for recent reproductive isolation between B. americanus and B. houstonensis. However, ABBA-BABA tests for ancient admixture indicated historic gene flow between Nearctic species while no signal of historic gene flow was detected between Nearctic and Middle-American species. We used an admixture analysis to recognize four Management Units (MU) based on observed genetic differentiation within B. houstonensis and recommend captive propagation, population supplementation, and habitat restoration efforts specific to each MU. Our results re-affirm the evolutionary novelty of an endangered relict.

Patterns of genetic divergence in the Rio Grande cooter (Pseudemys gorzugi), a riverine turtle inhabiting an arid and anthropogenically modified system.

The lower Rio Grande and Pecos River of the southwest United States of America have been heavily modified by human activities, profoundly impacting the integrity of their aquatic wildlife. In this context, we focused our study on the population genomics of the Rio Grande Cooter (Pseudemys gorzugi), a freshwater turtle of increasing conservation concern, residing in these two rivers and their tributaries. The genetic data revealed two distinct populations: one in the Pecos and Black rivers of New Mexico, and another in the Rio Grande and Devils River of Texas, with admixed individuals identified at the confluence of the Rio Grande and Pecos River. In addition to having a smaller geographic range, we found lower observed heterozygosity, reduced nucleotide diversity, and a smaller effective population size (Ne) in New Mexico population. Our results depict a significant isolation-by-distance pattern across their distribution, with migration being notably infrequent at river confluences. These findings are pivotal for future conservation and restoration strategies, emphasizing the need to recognize the unique needs of each population.

Mitochondrial sequence data reveal population structure within Pustulosa pustulosa.

Unionid mussels are among the most imperiled group of organisms in North America, and Pustulosa pustulosa is a freshwater species with a relatively wide latitudinal distribution that extends from southern Ontario, Canada, to Texas, USA. Considerable morphological and geographic variation in the genus Pustulosa (formerly Cyclonaias) has led to uncertainty over species boundaries, and recent studies have suggested revisions to species-level classifications by synonymizing C. aurea, C. houstonensis, C. mortoni, and C. refulgens with C. pustulosa (currently P. pustulosa). Owing to its wide range and shallow phylogenetic differentiation, we analyzed individuals of P. pustulosa using mitochondrial DNA sequence data under a population genetics framework. We included 496 individuals, which were comprised of 166 samples collected during this study and 330 additional sequences retrieved from GenBank. Pairwise ΦST measures based on ND1 data suggested there may be up to five major geographic groups present within P. pustulosa. Genetic differentiation between regions within Texas was higher compared to populations from the Mississippi and Great Lakes populations, which may reflect differences in historical connectivity. Mitochondrial sequence data also revealed varying demographic histories for each major group suggesting each geographic region has also experienced differential population dynamics in the past. Future surveys should consider exploring variation within species after phylogeographic delimitation has been performed. In this study, we begin to address this need for freshwater mussels via the P. pustulosa system.

Development and validation of an environmental DNA assay to detect federally threatened groundwater salamanders in central Texas.

The molecular detection of DNA fragments that are shed into the environment (eDNA) has become an increasingly applied tool used to inventory biological communities and to perform targeted species surveys. This method is particularly useful in habitats where it is difficult or not practical to visually detect or trap the target organisms. Central Texas Eurycea salamanders inhabit both surface and subterranean aquatic environments. Subterranean surveys are challenging or infeasible, and the detection of salamander eDNA in water samples is an appealing survey technique for these situations. Here, we develop and validate an eDNA assay using quantitative PCR for E. chisholmensis, E. naufragia, and E. tonkawae. These three species are federally threatened and constitute the Septentriomolge clade that occurs in the northern segment of the Edwards Aquifer. First, we validated the specificity of the assay in silico and with DNA extracted from tissue samples of both target Septentriomolge and non-target amphibians that overlap in distribution. Then, we evaluated the sensitivity of the assay in two controls, one with salamander-positive water and one at field sites known to be occupied by Septentriomolge. For the salamander-positive control, the estimated probability of eDNA occurrence (ψ) was 0.981 (SE = 0.019), and the estimated probability of detecting eDNA in a qPCR replicate (p) was 0.981 (SE = 0.011). For the field control, the estimated probability of eDNA occurring at a site (ψ) was 0.938 (95% CRI: 0.714-0.998). The estimated probability of collecting eDNA in a water sample (θ) was positively correlated with salamander relative density and ranged from 0.371 (95% CRI: 0.201-0.561) to 0.999 (95% CRI: 0.850- > 0.999) among sampled sites. Therefore, sites with low salamander density require more water samples for eDNA evaluation, and we determined that our site with the lowest estimated θ would require seven water samples for the cumulative collection probability to exceed 0.95. The estimated probability of detecting eDNA in a qPCR replicate (p) was 0.882 (95% CRI: 0.807-0.936), and our assay required two qPCR replicates for the cumulative detection probability to exceed 0.95. In complementary visual encounter surveys, the estimated probability of salamanders occurring at a known-occupied site was 0.905 (SE = 0.096), and the estimated probability of detecting salamanders in a visual encounter survey was 0.925 (SE = 0.052). We additionally discuss future research needed to refine this method and understand its limitations before practical application and incorporation into formal survey protocols for these taxa.

Quantification of chlamydiae in the endangered Houston toad (Anaxyrus houstonensis).

Two primer set/probe combinations targeting variable regions on the 23S rRNA gene were designed to detect and quantify chlamydiae in DNA extracted from brain swabs of the endangered Houston toad (Anaxyrus houstonensis) using SYBRGreen- and Taqman-based quantitative polymerase chain reaction (qPCR). Prevalence and abundance values for samples were generally different between SYBRGreen- and Taqman-based detection methods, with higher specificity observed for Taqman-based detection. Of the 314 samples analyzed, initial screening with SYBRGreen-based qPCR retrieved 138 positive samples, of which 52 were confirmed by Taqman-based analyses as chlamydiae. All of these samples were subsequently identified as Chlamydia pneumoniae by specific qPCR and confirmed by comparative sequence analyses of 23S rRNA gene amplicons. These results demonstrate the usefulness of our developed qPCR methods to screen for and verify prevalence of chlamydiae in DNA of brain swabs, and ultimately specifically identify and quantify chlamydiae, specifically C. pneumoniae in these samples.

Animal Species Recognition with Deep Convolutional Neural Networks from Ecological Camera Trap Images.

Accurate identification of animal species is necessary to understand biodiversity richness, monitor endangered species, and study the impact of climate change on species distribution within a specific region. Camera traps represent a passive monitoring technique that generates millions of ecological images. The vast numbers of images drive automated ecological analysis as essential, given that manual assessment of large datasets is laborious, time-consuming, and expensive. Deep learning networks have been advanced in the last few years to solve object and species identification tasks in the computer vision domain, providing state-of-the-art results. In our work, we trained and tested machine learning models to classify three animal groups (snakes, lizards, and toads) from camera trap images. We experimented with two pretrained models, VGG16 and ResNet50, and a self-trained convolutional neural network (CNN-1) with varying CNN layers and augmentation parameters. For multiclassification, CNN-1 achieved 72% accuracy, whereas VGG16 reached 87%, and ResNet50 attained 86% accuracy. These results demonstrate that the transfer learning approach outperforms the self-trained model performance. The models showed promising results in identifying species, especially those with challenging body sizes and vegetation.

Effects of salinization on the occurrence of a long-lived vertebrate in a desert river.

The lower Pecos River located in the southwest USA, is a naturally saline river system that has been significantly altered in relatively recent years. Climate change, coupled with anthropogenic disturbances such as dam construction have led to portions of the river becoming more susceptible to increased salinization and declines in water quality. These alterations have been documented to be detrimental to multiple freshwater communities; however, there is a lack of knowledge on how these alterations influence long-lived species in the river, such as freshwater turtles, where the effects can appear over dramatically different temporal scales. The Rio Grande Cooter (Pseudemys gorzugi) is a species of concern known to occur in the Pecos River. To understand the current distribution and habitat requirements for P. gorzugi in the Pecos River, we used a single-season, single-species occupancy modeling framework to estimate occurrence while accounting for the sampling process. Day of year, water surface area, and water visibility had the greatest influence on the ability to detect the species given a sampling unit is occupied. Conductivity (a measure of salinity) had the greatest influence on the occupancy probability for the species, where sites with higher conductivity coincided with lower occupancy probabilities. This study indicates that increased salinization on the lower Pecos River is a cause for concern regarding freshwater turtle populations within the Chihuahuan Desert.

Comparative host-pathogen associations of Snake Fungal Disease in sympatric species of water snakes (Nerodia).

The ascomycete fungus Ophidiomyces ophiodiicola (Oo) is the causative agent of ophidiomycosis (Snake Fungal Disease), which has been detected globally. However, surveillance efforts in the central U.S., specifically Texas, have been minimal. The threatened and rare Brazos water snake (Nerodia harteri harteri) is one of the most range restricted snakes in the U.S. and is sympatric with two wide-ranging congeners, Nerodia erythrogaster transversa and Nerodia rhombifer, in north central Texas; thus, providing an opportunity to test comparative host-pathogen associations in this system. To accomplish this, we surveyed a portion of the Brazos river drainage (~ 400 river km) over 29 months and tested 150 Nerodia individuals for the presence of Oo via quantitative PCR and recorded any potential signs of Oo infection. We found Oo was distributed across the entire range of N. h. harteri, Oo prevalence was 46% overall, and there was a significant association between Oo occurrence and signs of infection in our sample. Models indicated adults had a higher probability of Oo infection than juveniles and subadults, and adult N. h. harteri had a higher probability of infection than adult N. rhombifer but not higher than adult N. e. transversa. High Oo prevalence estimates (94.4%) in adult N. h. harteri has implications for their conservation and management owing to their patchy distribution, comparatively low genetic diversity, and threats from anthropogenic habitat modification.

Quantification of members of the Mycobacterium chelonae-abscessus complex in lesions of the endangered houston toad (Anaxyrus houstonensis).

Illumina-based 16S rRNA V3 amplicon sequencing of total DNA obtained from soft tissue lesions (joint granulomas) of the endangered Houston toad (Anaxyrus houstonensis) demonstrated that many reads represented members of the actinobacterial Mycobacterium chelonae-abscessus complex. In order to quantify members of this complex in those lesions, we designed three complex-specific primer set/probe combinations (sets I, II and III) targeting variable regions on the 23S rRNA gene for SybrGreen- and Taqman-based quantitative polymerase chain reaction (qPCR). Both SybrGreen- and Taqman-based analyses specifically detected members of the M. chelonae-abscessus complex in lesion samples, with numbers between 104 and 107 cells per 100-mg sample. Values within individual samples were generally comparable between SybrGreen- and Taqman-based detection methods and between all primer set/probe combinations, except for SybrGreen-based analyses of a few samples analyzed with primer set I that used a less specific forward primer. The development of highly specific detection and quantification methods for members of the M. chelonae-abscessus complex in lesion samples can enable group specific tracking of these organisms, particularly in captive or stewardship settings where source and transmission monitoring are valuable tools to husbandry and species conservation.

Predicting surface abundance of federally threatened Jollyville Plateau Salamanders (Eurycea tonkawae) to inform management activities at a highly modified urban spring.

Urban expansion has contributed to the loss of habitat for range restricted species across the globe. Managing wildlife populations within these urban settings presents the challenge of balancing human and wildlife needs. Jollyville Plateau Salamanders (Eurycea tonkawae) are a range restricted, federally threatened, species of neotenic brook salamander endemic to central Texas. Almost the entire geographic range of E. tonkawae is embedded in the Austin, Cedar Park, and Round Rock metropolitan areas of Travis and Williamson counties, Texas. Among E. tonkawae occupied sites, Brushy Creek Spring has experienced some of the most extensive anthropogenic disturbance. Today the site consists of small groundwater outlets that emerge in the seams within a concrete culvert underlying a highway. Salamanders persist within this system though they are rarely detected. Here, we model the occurrence of salamanders within the surface habitat of Brushy Creek Spring using generalized linear models. In the absence of available data regarding the amount of water that is discharged from the spring, we use accumulated rainfall as a proxy for discharge to estimate salamander abundance. Additionally, we present evidence of reproduction, recruitment, and subterranean movement by E. tonkawae throughout this site. Infrastructure maintenance is inevitable at Brushy Creek Spring. We intend for our results to inform when maintenance should occur, i.e., during environmental conditions when salamanders are less likely to be observed in the surface habitat, to avoid unnecessary impacts to this federally threatened species.

Environmental determinants of Batrachochytrium dendrobatidis and the likelihood of further dispersion in the face of climate change in Texas, USA.

One of the major drivers of amphibian population declines is Batrachochytrium dendrobatidis (Bd). We sought to identify the major environmental drivers of Bd prevalence in Texas, USA, by drawing results from museum specimens. We sampled one of the largest museum collections in Texas, the Biodiversity Research and Teaching Collections at Texas A&M University. Our sampling focused on the 9 amphibian species with the widest geographical distribution within the state, where we sub-sampled 30% of each species per decade from 1930 to present via skin swabs, totaling 1501 independent sampling events, and used quantitative real-time PCR (qPCR) to detect pathogen presence. We analyzed several geo-referenced variables describing climatic conditions to identify potential factors influencing the likelihood of presence of Bd using boosted regression trees. Our final model suggests the most influential variables are mean temperature of driest quarter, annual mean temperature, temperature annual range, and mean diurnal range. The most likely suitable range for Bd is currently found in the Blackland Prairie and Cross Timbers ecoregions. Results of our future (to the year 2040) projections suggest that Bd could expand its current distribution. Our model could play an important role when developing an integrated conservation plan through (1) focusing future field work on locations with a high likelihood of presence, (2) assisting in the choice of locations for restoration, and (3) developing future research plans including those necessary for projecting reactions to climate change. Our model also could integrate new presence data of Bd when they become available to enhance prediction precision.

Optimizing the power of human performed audio surveys for monitoring the endangered Houston toad using automated recording devices.

Knowledge regarding the locations of populations of endangered species is a critical part of recovery and facilitates land use planning that avoids unnecessary impacts. Regulatory agencies often support the development of survey guidelines designed to standardize the methods and maximize the probability of detection, thereby avoiding incorrectly concluding a species is absent from a site. Here, using simulations with data collected using automated recording devices (ARDs) we evaluated the efficacy of the existing U.S. Fish and Wildlife Service's survey requirements for the endangered Houston Toad (Anaxyrus houstonensis). We explored the effect of (1) increasing survey duration, (2) increasing the number of surveys, and (3) combinations of environmental conditions (e.g., temperature, humidity, rainfall) on the detection probability and the number of surveys needed to be 95% confident of absence. We found that increases in both the duration of the survey and the number of surveys conducted decreased the likelihood of incorrectly concluding the species was absent from the site, and that the number of surveys required to be 95% confident greatly exceeded the existing survey requirements. Targeting specific environmental conditions was also an effective way to decrease the number of surveys required but the infrequency in which these conditions occurred might make application difficult in some years. Overall, we suggest that the survey effort necessary to achieve confidence in the absence of Houston Toads at a site is more practically achievable with the use of ARDs, but this may not be suitable in all monitoring scenarios.

Influence of landscape condition on relative abundance and body condition of two generalist freshwater turtle species.

Anthropogenic land use changes have broad impacts on biological diversity, often resulting in shifts in community composition. While many studies have documented negative impacts on occurrence and abundance of species, less attention has been given to native species that potentially benefit from anthropogenic land use changes. For many species reaching high densities in human-dominated landscapes, it is unclear whether these environments represent higher quality habitat than more natural environments. We examined the influence of landscape ecological integrity on relative abundance and body condition of two native generalist freshwater turtle species that are prevalent in anthropogenic systems, the painted turtle (Chrysemys picta) and red-eared slider (Trachemys scripta elegans). Relative abundance was negatively associated with ecological integrity for both species, but the relationship was not strongly supported for painted turtles. Body condition was positively associated with ecological integrity for painted turtles, with no strong association for red-eared sliders. Our study suggests that both species benefitted at the population level from reduced ecological integrity, but individual-level habitat quality was reduced for painted turtles. The differing responses between these two habitat generalists could partially explain why red-eared sliders have become a widespread exotic invasive species, while painted turtles have not.

MYCOBACTERIA IN SKIN LESIONS AND THE HABITAT OF THE ENDANGERED HOUSTON TOAD (ANAXYRUS HOUSTONENSIS).

Head-starting of the federally endangered Houston toad (Anaxyrus houstonensis), that is, the release of egg strands, tadpoles, and metamorphic juveniles produced in captivity into the original breeding ponds, requires assessment of potential threats for the transmission of pathogens from captive to free-ranging toads. We used Illumina-based 16S rRNA V3 amplicon sequencing to investigate the community structure of bacteria from skin lesions of captive Houston toad and habitat (pond) samples. Proteobacteria, alone or together with Actinobacteria and, in some samples, Cyanobacteria represented virtually all reads in tissue lesion samples, whereas pond samples were much more diverse, with Acidobacteria, Actinobacteria, Bacteriodetes, Chloroflexi, Cyanobacteria, Firmicutes, Planctomycetes, Proteobacteria, and Verrucomicrobia present with little variation between samples. If present in lesions, Actinobacteria were largely represented by Mycobacteriaceae, and here mainly by one sequence identical to sequences of members of the Mycobacterium chelonae-abscessus complex. In pond samples, mycobacteria represented only a small portion of the actinobacteria, although at higher diversity with six distinct reads. Sequences for reads obtained from pond samples were identical to those representing the M. chelonae-abscessus complex, a group with Mycobacterium marinum, Mycobacterium kansasii, Mycobacterium avium, a group with Mycobacterium vaccae, Mycobacterium fortuitum, Mycobacterium poriferae, and a group with Mycobacterium elephantis and Mycobacterium celeriflavum, whereas sequences of high similarity were detected for reads related to those of Mycobacterium holsaticum, Mycobacterium pallens, and Mycobacterium obuense, and Mycobacterium goodii. Our results indicated that lesions observed on the Houston toad in captivity are not the result of mycobacteria in every case, and that the presence of mycobacteria in the captive colony does not represent a novel pathogen threat to the wild populations because such bacteria are also seen in the natural pond habitats for the Houston toad.

Morphometric data for five freshwater turtles in south, central, and west Texas.

From 2008 to 2013, we sampled freshwater turtle populations at 66 sites in south, central, and west Texas, USA. Sampling sites included ponds, lakes, resacas (oxbow lakes), canals, and rivers. We sampled turtle populations using baited hoop nets (66 sites) and basking traps (3 sites), and captured turtles by hand opportunistically in terrestrial habitat. We measured carapace length and width, plastron length and width, body depth, and weight of captured turtles. Excluding recaptures, we measured 356 Apalone spinifera emoryi (Texas Spiny Softshell), 24 Chelydra serpentina (Snapping Turtle), 20 Kinosternon flavescens (Yellow Mud Turtle), 47 Trachemys gaigeae (Big Bend Slider), and 1070 Trachemys scripta elegans (Red-eared Slider). Carapace length of Apalone spinifera emoryi ranged from 85 to 426 mm (mean = 182 mm). Carapace length of Chelydra serpentina ranged from 74 to 320 mm (mean = 233 mm). Carapace length of Kinosternon flavescens ranged from 64 to 147 mm (mean = 114 mm). Carapace length of Trachemys gaigeae ranged from 54 to 203 mm (mean = 141 mm). Carapace length of Trachemys scripta elegans ranged from 30 to 328 mm (mean = 171 mm). These data are useful for assessing spatial and temporal variation in size and body condition of freshwater turtles.

Evaluating the effects of red imported fire ants (Solenopsis invicta) on juvenile Houston Toads (Bufo [=Anaxyrus] houstonensis) in Colorado County, TX.

The spread of invasive species is considered a major threat to biodiversity, second only to habitat loss. Red Imported Fire Ants (Solenopsis invicta) are a globally invasive species with negative impacts reported on native invertebrate and vertebrate species. Federally endangered Houston Toads (Bufo [=Anaxyrus] houstonensis), endemic to Texas, are among the vertebrates reportedly negatively impacted by Red Imported Fire Ants (RIFA). Threats posed by RIFA to Houston Toads needed to be explicitly characterized. Large-scale chemical treatments to suppress RIFA and facilitate brood survival in Attwater's prairie-chickens (Tympanuchus cupido attwateri) at the Attwater Prairie Chicken National Wildlife Refuge (APCNWR) afforded us an opportunity to experimentally examine the influence of RIFA abundance on juvenile Houston Toad growth and survival. We also sought to examine whether juvenile Houston Toads could grow and survive in a vegetation type similar to a historic species locality. We conducted a terrestrial mesocosm experiment to test whether the application of bait-driven suppressant decreased counts of RIFA relative to untreated sites. We examined whether counts of native ant and non-ant native invertebrates were higher in response to potential decreases in RIFA. We compared growth and survival rates in juvenile Houston Toads among treated and untreated sites, expecting juvenile growth and survival to be higher in response to potentially decreased RIFA counts and increased native invertebrate counts. We saw lower counts of RIFA in treated prairies, but we also observed a decrease in native ant counts possibly due to chemical treatment. Therefore, the application of bait-driven suppressant may not affect RIFA alone. We saw no difference in counts of non-ant invertebrates among treated and untreated sites. Juvenile Houston Toads did not differ in growth and survival among treated and untreated sites. We recognize that the lack of a relationship between juvenile growth and survival with a treatment effect, and therefore RIFA abundance, may be limited to APCNWR. We encourage additional experimental studies to elucidate RIFA impacts at other sites. We extrapolated apparent survival estimates from our study to one year. These appear comparable to juvenile survivorship required in simulations for Houston Toad population persistence and on this basis, we recommend that APCNWR be re-evaluated as a reintroduction site for Houston Toads. We also recommend further studies to potentially broaden the regulatory definition of Houston Toad habitat beyond the current restrictive view of canopied forest alone. Such studies would need to examine the utility of native grasslands as dispersal corridors/upland habitat for juvenile Houston Toads. Our findings emphasize the utility of experimental studies in directly examining the influence of perceived threats to imperiled species and the role of such clarifications in adapting recovery efforts.

Detection of Salmonella in the intestine of Hypostomus plecostomus from the upper San Marcos River, Texas.

The prevalence of salmonellae in the intestines of the invasive suckermouth catfish Hypostomus plecostomus was assessed in the San Marcos River, just down-stream of its spring-fed headwaters. In 2014, H. plecostomus, sediment, and water samples were collected during 15 sampling events. A combination of semi-selective enrichment and quantitative polymerase chain reaction (qPCR) revealed the presence of salmonellae in 45% of the fish intestines across the entire year, with a prevalence range of 13-100% per sampling event. Repetitive element sequence-based PCR (rep-PCR) and multi-locus sequence typing (MLST) revealed a high diversity of salmonellae from fish intestine samples at individual sampling times, single or multiple presence of rep-PCR patterns and serotypes within individual fish, and identical rep-PCR patterns and serotypes for different fish within and across sampling events. Overall, 15 serotypes were identified by MLST, with a diversity range between one and seven serotypes per sampling event. Some serotypes were retrieved only once, while others were detected more frequently. A few serotypes were retrieved at several sampling times, nearly evenly distributed over the entire sampling period. Prevalence and diversity were independent of precipitation events, indicating the potential presence of environmental strains that are capable of long-term persistence in the environment.

Comparison of hoop-net trapping and visual surveys to monitor abundance of the Rio Grande cooter (Pseudemys gorzugi).

Abundance estimates play an important part in the regulatory and conservation decision-making process. It is important to correct monitoring data for imperfect detection when using these data to track spatial and temporal variation in abundance, especially in the case of rare and elusive species. This paper presents the first attempt to estimate abundance of the Rio Grande cooter (Pseudemys gorzugi) while explicitly considering the detection process. Specifically, in 2016 we monitored this rare species at two sites along the Black River, New Mexico via traditional baited hoop-net traps and less invasive visual surveys to evaluate the efficacy of these two sampling designs. We fitted the Huggins closed-capture estimator to estimate capture probabilities using the trap data and distance sampling models to estimate detection probabilities using the visual survey data. We found that only the visual survey with the highest number of observed turtles resulted in similar abundance estimates to those estimated using the trap data. However, the estimates of abundance from the remaining visual survey data were highly variable and often underestimated abundance relative to the estimates from the trap data. We suspect this pattern is related to changes in the basking behavior of the species and, thus, the availability of turtles to be detected even though all visual surveys were conducted when environmental conditions were similar. Regardless, we found that riverine habitat conditions limited our ability to properly conduct visual surveys at one site. Collectively, this suggests visual surveys may not be an effective sample design for this species in this river system. When analyzing the trap data, we found capture probabilities to be highly variable across sites and between age classes and that recapture probabilities were much lower than initial capture probabilities, highlighting the importance of accounting for detectability when monitoring this species. Although baited hoop-net traps seem to be an effective sampling design, it is important to note that this method required a relatively high trap effort to reliably estimate abundance. This information will be useful when developing a larger-scale, long-term monitoring program for this species of concern.

Observer-free experimental evaluation of habitat and distance effects on the detection of anuran and bird vocalizations.

Acoustic surveys of vocalizing animals are conducted to determine density, distribution, and diversity. Acoustic surveys are traditionally performed by human listeners, but automated recording devices (ARD) are becoming increasingly popular. Signal strength decays, or attenuates, with increasing distance between source and receiver and some habitat types may differentially increase attenuation beyond the effects of distance alone. These combined effects are rarely accounted for in acoustic monitoring programs. We evaluated the performance of three playback devices and three ARD models using the calls of six anurans, six birds, and four pure tones. Based on these evaluations, we determined the optimal playback and recording devices. Using these optimal devices, we broadcast and recorded vocalizations in five habitat types along 1,000 m transects. We used generalized linear models to test for effects of habitat, distance, species, environmental, and landscape variables. We predicted detection probabilities for each vocalization, in each habitat type, from 0 to 1,000 m. Among playback devices, only a remote predator caller simulated vocalizations consistently. Differences of ~10 dB were observed among ARDs. For all species, we found differences in detectability between open and closed canopy habitats. We observed large differences in predicted detection probability among species in each habitat type, as well as along 1,000 m transects. Increases in temperature, barometric pressure, and wind speed significantly decreased detection probability. However, aside from differences among species, habitat, and distance, topography impeding a line-of-sight between sound source and receiver had the greatest negative influence on detections. Our results suggest researchers should model the effects of habitat, distance, and frequency on detection probability when performing acoustic surveys. To optimize survey design, we recommend pilot measurements among varying habitats.

Effects of nitrogenous wastes on survival of the Barton Springs salamander (Eurycea sosorum).

The objective of our study was to determine the acute toxicity of 3 common aquatic nitrogenous toxicants to the federally endangered Barton Springs salamander (Eurycea sosorum). Based on our findings, the 96-h median lethal concentrations (96-h LC50) for un-ionized ammonia-N, nitrite-N, and nitrate-N to E. sosorum are 2.0 ± 0.32, 31.7 ± 4.02, and 968.5 ± 150.6 mg/L, respectively. These results establish a benchmark for the tolerance of plethodontid salamanders to these toxicants and indicate that current water quality criteria are adequate for their protection. Environ Toxicol Chem 2017;36:3003-3007. © 2017 SETAC.