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 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.

Specimen collection is essential for modern science.

Natural history museums are vital repositories of specimens, samples and data that inform about the natural world; this Formal Comment revisits a Perspective that advocated for the adoption of compassionate collection practices, querying whether it will ever be possible to completely do away with whole animal specimen collection.

PREVALENCE OF RANAVIRUS IN SPOTTED SALAMANDER (AMBYSTOMA MACULATUM) LARVAE FROM CREATED VERNAL POOLS IN WEST VIRGINIA, USA.

Ranavirosis is a disease of high concern for amphibians due to widespread documentation of its lethal and sublethal impacts and its high transmission potential across populations and species. We investigated whether spotted salamander (Ambystoma maculatum) ranavirus prevalence and viral load were associated with habitat characteristics, genetic diversity, corticosterone levels, and body size. In 2015 and 2016, we sampled 34 recently created vernal pools in the Monongahela National Forest, West Virginia, USA. We collected tail clippings from 1,128 spotted salamander larvae and waterborne hormone samples from 436 of those larvae, along with eight environmental characteristics of the pools. Over the 2-yr period, we detected ranavirus in 62% of pools, with prevalence ranging from 0% to 63% (mean, 7.68%). Spotted salamander size was positively correlated with ranavirus presence and viral load; however, we did not find associations between ranavirus prevalence or viral load and habitat characteristics, spotted salamander genetic diversity, relatedness, effective number of breeders, or corticosterone levels. The widespread occurrence of ranavirus in the vernal pools illustrates the potential for rapid natural introduction of the pathogen to created wetlands. Managers could consider monitoring local distributions of ranavirus before creation of new vernal pools to guide strategic placement of the wetlands to minimize occurrence and prevalence of this pathogen.

Development and validation of an eDNA protocol for monitoring endemic Asian spiny frogs in the Himalayan region of Pakistan.

Wildlife monitoring programs are instrumental for the assessment of species, habitat status, and for the management of factors affecting them. This is particularly important for species found in freshwater ecosystems, such as amphibians, as they have higher estimated extinction rates than terrestrial species. We developed and validated two species-specific environmental DNA (eDNA) protocols and applied them in the field to detect the Hazara Torrent Frog (Allopaa hazarensis) and Murree Hills Frog (Nanorana vicina). Additionally, we compared eDNA surveys with visual encounter surveys and estimated site occupancy. eDNA surveys resulted in higher occurrence probabilities for both A. hazarensis and N. vicina than for visual encounter surveys. Detection probability using eDNA was greater for both species, particularly for A. hazarensis. The top-ranked detection model for visual encounter surveys included effects of both year and temperature on both species, and the top-ranked occupancy model included effects of elevation and year. The top-ranked detection model for eDNA data was the null model, and the top-ranked occupancy model included effects of elevation, year, and wetland type. To our knowledge, this is the first time an eDNA survey has been used to monitor amphibian species in the Himalayan region.

A drone-based survey for large, basking freshwater turtle species.

Conservation concerns are increasing for numerous freshwater turtle species, including Pseudemys gorzugi, which has led to a call for more research. However, traditional sampling methodologies are often time consuming, labor intensive, and invasive, restricting the amount of data that can be collected. Biases of traditional sampling methods can further impair the quality of the data collected, and these shortfalls may discourage their use. The use of unmanned aerial vehicles (UAVs, drones) for conducting wildlife surveys has recently demonstrated the potential to bridge gaps in data collection by offering a less labor intensive, minimally invasive, and more efficient process. Photographs and video can be obtained by camera attachments during a drone flight and analyzed to determine population counts, abundance, and other types of data. In this study we developed a detailed protocol to survey for large, freshwater turtle species in an arid, riverine landscape. This protocol was implemented with a DJI Matrice 600 Pro drone and a SONY ILCE α6000 digital camera to determine P. gorzugi and sympatric turtle species occurrence across 42 sites in southwestern Texas, USA. The use of a large drone and high-resolution camera resulted in high identification percentages, demonstrating the potential of drones to survey for large, freshwater turtle species. Numerous advantages to drone-based surveys were identified as well as some challenges, which were addressed with additional refinement of the protocol. Our data highlight the utility of drones for conducting freshwater turtle surveys and provide a guideline to those considering implementing drone-mounted high-resolution cameras as a survey tool.

SEASONALITY IN BATRACHOCHYTRIUM DENDROBATIDIS DETECTION IN AMPHIBIANS IN CENTRAL OKLAHOMA, USA.

Chytridiomycosis, an infectious disease caused by the fungus Batrachochytrium dendrobatidis (chytrid or Bd), has not been well studied in Oklahoma. This is of particular concern regarding the connection between seasonality and chytrid infection. To further investigate this connection, chytrid prevalence and infection load were quantified within amphibians in central Oklahoma from March to October, across two sites in Oklahoma Co. and two sites in Cleveland Co. The results show a trend between seasonality and chytrid, with spring and fall showing higher prevalence and summer showing lower prevalence, which coincides closely with the preferred chytrid growth temperatures. Additionally, periods of high rainfall in May 2015 are linked to increased chytrid prevalence, as has been suggested by other research. Additionally, species exhibiting high chytrid prevalence follow the results of previous studies: Blanchard's cricket frog (Acris blanchardi), American bullfrog (Rana catesbeiana), and southern leopard frog (Rana sphenocephala).

Water-borne and plasma corticosterone are not correlated in spotted salamanders.

Water-borne hormone measurement is a noninvasive method suitable for amphibians of all sizes that are otherwise difficult to sample. For this method, containment-water is assayed for hormones released by the animal. Originally developed in fish, the method has expanded to amphibians, but requires additional species-specific validations. We wanted to determine physiological relevance of water-borne corticosterone in spotted salamanders (Ambystoma maculatum) by comparing concentrations to those taken using established corticosterone sampling methods, such as plasma. Using a mixture of field and laboratory studies, we compared water-borne corticosterone levels to other traditional methods of sampling corticosterone for spotted salamander larvae, metamorphs, and adults. Despite multiple attempts, and detecting differences between age groups, we found no correlations between water-borne and plasma corticosterone levels in any age group. Water-borne sampling measures a rate of release; whereas plasma is the concentration circulating in the blood. The unique units of measurement may inherently prevent correlations between the two. These two methods may also require different interpretations of the data and the physiological meaning. We also note caveats with the method, including how to account for differences in body size and life history stages. Collectively, our results illustrate the importance of careful validation of water-borne hormone levels in each species in order to understand its physiological significance.

Concurrent Infection of Batrachochytrium dendrobatidis and Ranavirus among Native Amphibians from Northeastern Oklahoma, USA.

Global amphibian decline continues to be a great concern despite our increased understanding of the causes behind the observed patterns of the decline, such as habitat modification and infectious diseases. Although there is a large body of literature on the topic of amphibian infectious diseases, pathogen prevalence and distribution among entire communities of species in many regions remain poorly understood. In addition to these geographic gaps in our understanding, past work has focused largely on individual pathogens, either Batrachochytrium dendrobatidis (Bd) or ranavirus (RV), rather than dual infection rates among host species. We sampled for prevalence and infection load of both pathogens in 514 amphibians across 16 total sites in northeastern Oklahoma. Amphibians were caught by hand, net, or seine; they were swabbed to screen for Bd; and liver tissue samples were collected to screen for RV. Overall results of quantitative PCR assays showed that 7% of screened individuals were infected with RV only, 37% were infected with Bd only, and 9% were infected with both pathogens simultaneously. We also documented disease presence in several rare amphibian species that are currently being monitored as species of concern due to their small population sizes in Oklahoma. This study synthesizes a growing body of research regarding infectious diseases among amphibian communities in the central United States.

Corrigenda: Davis DR, LaDuc TJ (2018) Amphibians and reptiles of C. E. Miller Ranch and the Sierra Vieja, Chihuahuan Desert, Texas, USA. ZooKeys 735:97-130. https://doi.org/10.3897/zookeys.735.22200.

[This corrects the article DOI: 10.3897/zookeys.735.22200.].

Amphibians and reptiles of C. E. Miller Ranch and the Sierra Vieja, Chihuahuan Desert, Texas, USA.

We report the occurrence of 50 species of amphibians and reptiles recently collected on C. E. Miller Ranch and the Sierra Vieja in the Chihuahuan Desert of Texas, USA and describe their perceived distribution and abundance across various habitat associations of the region. Our recent surveys follow intense, historic sampling of amphibians and reptiles from this region in 1948. Of the 50 species detected in recent surveys, six were not collected in 1948 and an additional three species documented in 1948 have yet to be detected in a 14-year period of recent surveys. Combining data from both historic and recent surveys, a total of 53 species of amphibians and reptiles are known from the ranch (11 amphibians, 42 reptiles). Land stewardship and conservation practices have likely contributed to the persistence of the majority of these species through time. Additionally, we discuss the status of amphibians and reptiles not collected during recent surveys and comment on potential species that have not yet been detected.

A new species of Pseudogekko (Squamata: Gekkonidae) from the Romblon Island Group, Central Philippines.

We describe a new species of lizard in the genus Pseudogekko from Sibuyan and Tablas islands in the Romblon Island Group of the central Philippines. The new species is diagnosed from other Philippine Pseudogekko by body size and shape, color pattern, and multiple differences in scale characteristics. Pseudogekko isapa sp. nov. has been collected only twice from leaves of shrubs in forested habitat on Sibuyan and Tablas islands. The distinctive new species of false gecko is undoubtedly endemic to this single, isolated island group. The fact that populations of such a distinctive new species of Pseudogekko has escaped notice of herpetologists on the reasonably well-studied and largely protected Sibuyan Island further emphasizes the secretive and forest-dependent habits of Philippine false geckos. These characteristics of their behavior and natural history render them difficult to study and challenge biologists' efforts to accurately assess their conservation status.

Additions to Philippine Slender Skinks of the Brachymeles bonitae Complex (Reptilia: Squamata: Scincidae) I: a new species from Lubang Island.

A new species of slender skink is described from the Philippines. The species is endemic to Lubang Island, and is assigned to the Brachymeles bonitae Complex based on phenotypic and genetic data. Specimens were collected from Lubang Island between 1991 and 2012, and were examined based on morphological data (qualitative traits, meristic counts, and mensural measurements). Published genetic sequence data from phylogenetic studies of the genus reveal the new species to be highly divergent from congeners. Brachymeles ligtas sp. nov. is differentiated from other members of the genus based on a number of distinct morphological features, including small body size (SVL 60.7-79.6 mm), bidactyl fore-limbs, digitless hind limbs, high number of presacral vertebrae (50), and the absence of auricular openings. Additionally, the new species has diagnostic, distinct dorsal head scale patterns. This new species becomes the only member of the genus known to occur on the deep-ocean island of Lubang.

Additions to Philippine Slender Skinks of the Brachymeles bonitae Complex (Reptilia: Squamata: Scincidae) II: a new species from the northern Philippines.

We describe a new digitless scincid lizard of the genus Brachymeles from northern Luzon and Camiguin Norte islands in the Philippines. This species belongs to the Brachymeles bonitae Complex, and both molecular and morphological data confirm that this species is distinct from all other congeners. Formerly considered to be a single widespread species, this group of species has been the focus of recent systematic reviews. Here we describe a new species in the B. bonitae Complex, recognized currently to constitute five species. Brachymeles ilocandia sp. nov. is the second digitless and the seventeenth non-pentadactyl species in genus. The description of this species brings the total number of species in the genus to 40, and provides new insight into unique distribution patterns of species of the northern Philippines.

Additions to Philippine Slender Skinks of the Brachymeles bonitae Complex (Reptilia: Squamata: Scincidae) III: a new species from Tablas Island.

Studies of the diversity of Philippine amphibians and reptiles have resulted in the continued description of cryptic species. Species formerly thought to range across multiple recognized faunal regions are now considered to be assemblages of multiple unique species, each restricted to a single faunal region. This pattern continues to hold true when considering Philippine skinks of the genus Brachymeles. Recent studies have resulted in the description of numerous unique species with many exhibiting various degrees of digit loss or limb reduction, as well as suggesting that unique lineages are still present in the B. bonitae Complex. In this paper, we describe a new species of fossorial skink within this species complex from Tablas Island based on collections made nearly 50 years ago. Although no genetic data are available for the new species, examinations of morphological data (qualitative traits, meristic counts, and mensural measurements) support its distinction from all other members of the genus. Brachymeles dalawangdaliri sp. nov. is differentiated from other members of the genus based on a suite of unique phenotypic characteristics, including a small body size (SVL 66.0-80.9 mm), bidactyl fore-limbs, digitless, unidactyl, or bidactyl hind limbs, a high number of presacral vertebrae (49), the absence of auricular openings, and distinct dorsal head scale patterns. The description of the new species increases the diversity of endemic vertebrates recognized to occur in the Romblon Island Group in the central Philippines.

Redescription of the rare Philippine false gecko Pseudogekko brevipes (Reptilia: Squamata: Gekkonidae) and description of a new species.

Recent investigations into the species diversity of false geckos (genus Pseudogekko Taylor) have revealed several cryptic species, highlighting the need for a more thorough understanding of diversity within this enigmatic genus of endemic Philippine geckos. Newly available genetic data reveal that two of the four currently recognized species are complexes of multiple deeply divergent evolutionary lineages. In this paper we evaluate species diversity in one of these complexes, P. brevipes Boettger, and describe one additional new species. For nearly a century, P. brevipes has been recognized as a single, "widespread" species with a geographic range spanning two major faunal regions and several island groups. Poor understanding of this species has persisted due to both limited sampling and its apparent rarity. We evaluate both morphological and genetic data to define species limits in P. brevipes, and find character-based evidence to justify the recognition of two unique evolutionary lineages, one of which we describe as a new species (P. atiorum sp. nov.). The species included in this study have allopatric distributions and differ from congeners by numerous diagnostic characters of external morphology, and therefore should be recognized as full species in accordance with lineage-based species concepts. This newly described species increases the total number of species of Pseudogekko to seven.

Behavioral and physiological antipredator responses of the San Marcos salamander, Eurycea nana.

Exposure to predatory stimuli typically results in the elevation of circulating glucocorticoid levels and a behavioral response of freezing or escape behavior in many prey species. Corticosterone (CORT) is the main glucocorticoid in amphibians and is known to be important in modulating many behaviors and developmental functions. The federally threatened San Marcos salamander, Eurycea nana, decreases activity in response to both native and introduced predatory fish, however, experience may further influence these interactions. To better understand the indirect effects of fish predators on this salamander, we examined both the antipredator behavior and water-borne CORT release rates in response to chemical cues (kairomones) from two fish species that varied in temporal risk of predation: (1) a low encounter frequency predator (largemouth bass, Micropterus salmoides), (2) a high encounter frequency predator (redbreast sunfish, Lepomis auritus), and (3) a blank water control. Salamanders reduced activity (antipredator response) after exposure to both predator treatments, but not to the blank water control, and the response to M. salmoides was significantly stronger than that to L. auritus. The CORT response (post-stimulus/pre-stimulus release rates) did not differ between the blank water control and L. auritus treatments, and both were significantly less than the CORT response to M. salmoides. Overall, E. nana showed a decreased antipredator response and no CORT response towards the high encounter frequency L. auritus as compared to the low encounter frequency M. salmoides. Eurycea nana may mute antipredator and CORT responses to high temporal frequency predators. There was, however, no correlation between CORT release rates and antipredator behavior, which suggests that the presence of predators may be affecting CORT response and behavior independently.

Cryptic diversity and population genetic structure in the rare, endemic, forest-obligate, slender geckos of the Philippines.

Recent studies of forest lizards in Southeast Asia have highlighted spectacular morphological and cryptic genetic diversity in several poorly known clades. Unfortunately, many of the included species have microhabitat preferences for forested environments, and therefore they are threatened by extensive forest destruction throughout the region. This is particularly true in the Philippines, an archipelago with a strikingly high proportion (84%) of endemic geckos. Abundances inferred from historical museum collections suggests that we are in a critical period where apparent declines in population viability and species' abundance have taken place faster than the growth in our understanding of alpha diversity. This phenomenon is exemplified in the exceedingly rare Philippine slender forest geckos of the genus Pseudogekko. Most of the known species are rarely encountered by field biologists, and species boundaries are unclear; this poor state of knowledge impedes effective conservation measures. Using the first multilocus phylogeny for these taxa, and phylogenetic and population genetic approaches, we elucidate evolutionary lineages and delimit species-level conservation targets in this unique radiation of endemic Philippine geckos. The results support the presence of widespread cryptic diversity in the genus, providing a framework for the re-evaluation of conservation priorities aimed at protecting these rare, forest-obligate species.