During recent surveys in Gaoligongshan Mountain Range, Yunnan Province of China, we collected specimens of Scutiger from a montane stream area at an elevation of 3000 m. Subsequent study based on morphological, molecular, and acoustic data reveals that the newly collected specimens represent an unknown taxon and is described herein: Scutiger tengchongensis, new species. The new species can be distinguished from its congeners by a combination of the following characters: (1) relatively small size (adult male SVL 36.0–40.1 mm, n = 8); (2) vomerine teeth absent; (3) dorsum with large prominent, conical-shaped and longitudinal tubercles; (4) large tubercles on dorsum covered by numerous tiny black spines in males in breeding condition; (5) a pair of pectoral glands and a pair of axillary glands present on the chest, and covered by dense tiny black spines in males in breeding condition; (6) pectoral gland about twice the size of axillary gland; (7) inner three fingers with black nuptial spines in males in breeding condition; (8) nuptial spines on first and second fingers larger than those on third finger; (9) black spines on the belly absent; (10) vocal sac absent; and (11) toe webbing rudimentary. The discovery of S. tengchongensis, new species, represents the southernmost record of the genus worldwide, and the new species is the smallest known member of the genus in body size.

Cognition and learning have been widely studied in vertebrates, but not across much phylogenetic breadth. Non-avian reptiles, for example, have been poorly studied. Anecdotal observations and a few previous studies suggest that lizards may have strong cognitive skills owing, in part, to behaviors such as optimal foraging and territoriality. We tested four lizard species, including three species of monitor lizard (Varanus spp.) and one species of beaded lizard (Heloderma), in a longitudinal, repeated-trials experimental design using a puzzle-feeder device to evaluate learning, in the form of latency trends over time. We used a Bayesian multilevel modeling statistical method and incorporated unsuccessful trials as censored data. Collectively, all lizards showed a pattern of decreasing latencies over time. We interpret this pattern as learning among our lizards. Notable individual and inter-specific differences were evident, however, suggesting that learning abilities differed among the lizards. In this case, the monitor lizards exhibited steeper declines in latencies and greatly reduced inter-individual variation in comparison to the beaded lizards. Finally, we found differential use of the claws versus the snout among the lizards, which is consistent with a previously posed hypothesis based on different species than we measured.

Channa lipor, a new species of snakehead of the C. gachua species-group, is described based on 11 specimens from Meghalaya, Northeast India. It is distinguished from its congeners by possessing an orange, bronze-brown dorsum and fins, 9–12 black spots or blotches on the dorsal-fin sub-margin appearing parallel along the length of the dorsal-fin base, six oblique brown bars on the upper half on the flank, presence of seven gray to brown zigzag bands on the caudal fin, and fewer anal-fin rays. Channa lipor, new species, morphologically resembles C. aurantipectoralis, but the partial cox1 gene sequences reveal a genetic distance of 12.6–13.1% between them, and Channa lipor, new species, also possesses deep sequence divergence from any known populations of C. gachua. It differs from the topotypic C. gachua by having fewer anal-fin rays (20 vs. 22–24), fewer dorsal-fin rays (29–32 vs. 34–36), and fewer pectoral-fin rays (12–14 vs. 15–17), and in tooth pattern, by having the fifth ceratobranchial curved with four rows of teeth, outer row with 11 thick teeth; palatine with three rows of curved teeth, inner row with 14 large inwardly curved teeth; and dentary with long canine-like teeth in the posterior end.

Sea turtle hatchlings are known to vocalize; however, the purpose of these vocalizations is currently unknown. One hypothesis is that these vocalizations serve to synchronize hatching or starting the emergence from the nest. To test this hypothesis, we characterized and compared the frequency and duration of the sounds made by Olive Ridley Turtles (Lepidochelys olivacea) during incubation, hatching, and emerging from the nest. We also determined whether hatchlings continue to vocalize after emerging from the nest. Lastly, we investigated species-specific variations in the vocalizations of Olive Ridley, Leatherback (Dermochelys coriacea), and Green (Chelonia mydas) turtle hatchlings. Sound production was more frequent during incubation than during hatching or emerging from the nest, and in two of the three nests, a unique ‘pulse’ sound was heard during incubation. However, there were no significant differences in the frequency range or duration of the vocalizations between incubation, hatching, and emerging from the nest. Furthermore, hatchlings were recorded and continued to vocalize after emerging from the nest. From this we conclude it is unlikely that the sounds made by the hatchlings help to synchronize hatching or emergence behavior. Instead, they are likely by-products of other processes, such as embryo movement, serving little biological purpose. Nevertheless, we recommend further research to unequivocally confirm this is the case, and it remains highly interesting that sound production is a universal trait among cheloniid species.

Oviducts of paedomorphic and metamorphic Ambystoma talpoideum were examined to determine if previously observed differences in egg mass characteristics between the two morphs originated from differences in oviductal histomorphology/histochemistry. We found no evidence to support this hypothesis. At least seven oviductal regions were easily identified in the oviducts of both paedomorphic and metamorphic individuals; these were as follows (from cranial to caudal): 1) pars recta (termed Region A), 2) pars convoluta (termed Regions B through F), and 3) the uterine region (termed Region G). While slight structure and staining differences were evident in Region D of the pars convoluta when paedomorphic A. talpoideum from South Carolina were compared with metamorphic A. talpoideum from Arkansas, there were no differences observed between paedomorphic and metamorphic individuals of A. talpoideum from Arkansas. Thus, any observed oviductal variation was most likely the result of geographic variation and not from differences involving metamorphosis between paedomorphic and metamorphic A. talpoideum. The number of oviductal regions of A. talpoideum was comparable with previous studies on plethodontids and salamandrids, and this study represents the first treatise of oviductal anatomy of an ambystomatid using a histochemical approach beyond general staining with hematoxylin and eosin.

African snake-eaters of the genus Polemon are cryptic, fossorial snakes that mainly inhabit the forests of central, eastern, and western Africa. Molecular results from a previous study demonstrated that Polemon christyi is not monophyletic—two distinct lineages were recovered from Uganda (the type locality) and southeastern Democratic Republic of the Congo (DRC). Genetic data indicated differences in sequence divergence and encoded amino acids between these lineages. Based on these molecular differences and diagnostic differences in morphology, we describe the lineage from southeastern DRC as a new species. Literature records indicate that it likely occurs in adjacent Tanzania and Zambia. It is the first species of Polemon to be described in over 70 years.

Road mortality has become a primary threat to turtle populations. A variety of strategies have been applied to reduce turtle mortality; however, the effectiveness of these solutions remains largely untested. The few tests that have been conducted have often found the mitigation actions to be ineffective or prohibitively expensive for implementation at a landscape level. Thus, there is a need for inexpensive, effective, and rigorously tested methods for reducing turtle mortality on roads. Here, we investigate the potential efficacy of an inexpensive mitigation strategy, consisting of a barrier wall fabricated from High Density Polyethylene (HDPE) half-pipe, partnered with a pre-existing semiaquatic HDPE culvert as an ecopassage. We used survey data and a Before-After-Control-Impact (BACI) experimental design to assess the effectiveness of the mitigation and quantified the use of the ecopassage at the impact site after the barrier wall was installed. We found a significant interaction between the treatment effect and time period, with the impact site (but not control sites) showing significantly lower turtle counts after mitigation. The barrier wall reduced the number of turtles accessing the road by 94%, and there was no evidence of turtles circumnavigating the barrier. Turtles used the pre-existing culvert to pass under the road. We conclude that HDPE half-pipe partnered with an existing semiaquatic HDPE culvert can reduce road access by, and road mortality of, turtles. The use of standard road construction materials for barrier walls and underpasses results in a cost-effective design scalable to a landscape level.

Quinca mirificaMees, 1966 is a relatively large cardinalfish endemic to western Australia that displays a combination of unusual traits for an apogonid, including solitary behavior, conspicuous ephemeral sexual dichromatism, lack of a dispersing larval period, and production of non-functional oocytes. The ovaries are of cystovarian type and present a germinal ridge running longitudinally along the entire medial face. Ovaries show distinct division in oocyte composition, with a clear division between a rostral section composed of normal oocytes, and a caudal section that is composed of a mass of ∼13,000 non-vitellogenic oocytes of ∼0.2–0.3 mm in diameter, which after hydration, measure ∼0.8–1.0 mm. The egg clutch contains ∼200–250 eggs of ∼3.5 mm in diameter. Egg clutches sometimes break during the transfer, and males retrieve no more than ∼50% of the eggs. Courtship and mating include a “holding behavior” not observed in other apogonids, and the mating process (egg transfer) lasts up to about 90 seconds, the longest observed in any apogonid. Males (orally) incubate the eggs and hatched embryos for about 22–26 days, fasting during this period. Embryos hatch after ∼22 days (8.5 mm SL) and remain in the male's oral cavity for another four days feeding endogenously before being released as juveniles (SL = 9.5 mm). Females became sexually mature at about 10–11 months of age (∼77 mm SL). A study with two females (13 spawns) showed mean frequency of ∼33 days between mating events. Gross anatomy of gonads, embryology, and juvenile development are described.

Because of their small size and fragility, larvae of many amphibian species are difficult to mark unambiguously for experimental studies. This constraint limits our ability to run experiments in which individual interactions matter and so limits our ability to study how social behaviors affect individual fitness. We anesthetized small (<8 mm long) Cane Toad (Rhinella marina) tadpoles and used a biopsy punch to remove tissue from either the dorsal or ventral tail fin, enabling the identification of several cohorts within an enclosed system. The tail fin clips remained readable for >30 days and had only minimal effects on tadpole growth, survival, and swimming performance. Marking the membrane of a tadpole tail fin provides a cost-effective method for cohort marking with minimal equipment and training. It also provides the opportunity to gather extra data on an individual (such as DNA) with minimal further stress to the animal.

The ability to identify individuals is important for the success of many behavioral and ecological studies. In fishes, there is a lot of variation in body size, shape, skin thickness, behavior, and ecology, which means that any given marking/ tagging method may not work well for all species. For the Humbug Damselfish Dascyllus aruanus, a widely used model species of coral-reef fish, we found that standard tagging methodologies (e.g., phenotypic variation, beads and PVC tags, fluorescent elastomer injections) were ineffective as they could not be applied to the fish or easily detected by observers or from videos. In response, we developed a new method to temporarily tag D. aruanus using colored plastic films and topical surgical glue. These films were easily detectable both directly by human observers and indirectly by video/photo-cameras. We tested the efficacy of this new method by examining the survival time of the tags at various positions on the body. Our results showed that the optimal tag locations were dorsal anterior (with a median attachment time of 53 hours) and dorsal middle (with a median attachment time of 49 hours). Total length of fish was not a significant predictor of tag life. In sum, we demonstrate an effective new method for temporarily tagging a widely studied coral reef fish. This method could be applied to other fishes and aquatic organisms (e.g., amphibians) in both marine and freshwater ecosystems.

Successful colonization by an invasive species is often associated with flexibility in life-history traits. The number and timing of reproductive events are among the most important of those life-history traits and are often strongly influenced by habitat stability. An invasive fish, Brook Stickleback (Culaea inconstans), was first detected in eastern Washington in 1999. They have subsequently expanded their range into water bodies of variable habitat stability, i.e., permanent, semi-permanent, and ephemeral water bodies. We hypothesized that Brook Stickleback living in unstable and unpredictable habitats would exhibit “fast” life-history traits relative to Brook Stickleback living in more stable habitats. Brook Stickleback were collected from 18 water bodies and standard length was measured. Otoliths were then extracted to determine age. Average age of Brook Stickleback in permanent wetlands was significantly greater than that of fish in semi-permanent and ephemeral wetlands, but there was no significant difference in age structure between semi-permanent and ephemeral wetlands (permanent x̄ = 1.24, semi-permanent x̄ = 0.73, ephemeral x̄ = 0.76, P < 0.001). Average length at age class 0+ was significantly less than age classes I, II, and III; however, there was no significant difference between age classes I, II, and III (0+: x̄= 39.73; I: x̄ = 46.42; II: x̄= 45.14; III: x̄= 49.10; P < 0.001). The results suggest that Brook Stickleback exhibit different life-history characteristics depending upon the habitats in which they are found. Brook Stickleback in unstable habitats may be functionally semelparous; however, it is possible that some individuals may produce multiple clutches within a single spawning season. Finally, we found that the only accurate way to measure Brook Stickleback age is through otolith analysis, drawing into question the conclusions of previous work in which length was used as a surrogate for age.

Gut microbiota of vertebrate hosts play crucial roles for the host they inhabit. In this study, we compared the composition and predicted functions of gut microbiota of female individuals belonging to four frog species from different habitats using bacterial 16S rRNA gene sequencing. Our results showed that the gut microbiota of the examined frog species were dominated by the phyla Firmicutes, Proteobacteria, Bacteriodetes, Verrucomicrobia, and Fusobacteria. There were significant differences in the relative abundance of dominant phyla among the four species, although the relative abundance of dominant phyla in the microbiota of individuals of Odorrana tormota was more evenly balanced (ranged from 10.45% for Fusobacteria to 27.21% for Bacteroidetes). The alpha diversity patterns varied depending on the microbial taxon level. At the phylum level, the Chao1 diversity was positively associated with body weight, with the diversity index scores of O. tormota and Amolops wuyiensis markedly lower than those of Odorrana schmackeri and Polypedates megacephalus. At the genus level, Chao1 diversity was associated with evolutionary relationships among species, with the Chao1 diversity index score for P. megacephalus of distant evolutionary relationships being significantly higher than the scores for frogs with close evolutionary relationships. Moreover, permutational multivariate analysis of variance revealed significant differences in the gut bacterial community structure among species (R² = 0.510, P = 0.001). Further, the predicted functions of the gut microbiota were similar for species with close evolutionary relationships but differed with distant evolutionary relationships. Taken together, our results indicate that wild frogs have species-specific microbial communities and provide insight into the ecological and evolutionary dynamics of host–microbe interactions.

Physiological responses to stress are important indicators of the effects of environmental disruption on individuals and, therefore, a way to determine the health of populations. These responses can be measured in a variety of ways, including the survey of differential counts of white blood cells. Baseline cell numbers and neutrophil to lymphocyte ratios have been determined for a number of species in the genus Ambystoma, but not for A. tigrinum, and baseline values are necessary for assessments of stress in natural and captive populations. We counted white blood cells in blood smears from long-term captive and recently captured Eastern Tiger Salamanders (A. tigrinum) and compared the proportions of each cell type between these two samples. We also compared our results to the published values for other post-metamorphic or paedomorphic ambystomatids. Mean neutrophil to lymphocyte ratios, a measure of stress, were higher in our captive salamanders (0.81) than in the wild sample (0.41), as were the mean number of basophils (36.0 for captive and 10.3 for wild). The cell counts for our wild salamanders were comparable to those for other unstressed ambystomatids. Our results suggest that our long-term captive salamanders are under a small degree of stress and are not a good source of baseline values for this species.

Organisms that experience large changes in body size during the life span often exhibit differences in resource use among life stages. Ontogenetic shifts in habitat use reduce intraspecific competition and predation and are common in lotic organisms. Although information on the immature life stages of the Hellbender (Cryptobranchus alleganiensis) is limited, this aquatic salamander exhibits ontogenetic shifts in habitat use in some streams, with adults sheltering under large rocks and larvae utilizing interstitial spaces of gravel beds. Due to the geomorphology of Little River, Tennessee, however, limited interstitial spaces within the gravel are often filled with sand. Therefore, we quantified microhabitat parameters for three life stages of Hellbenders (larvae, sub-adult, adult) to determine if an ontogenetic shift in microhabitat occurred in Little River. We found no significant differences in stream substrate at capture sites among the stages, but there was a positive correlation between rock shelters underlain with very coarse gravel and overall Hellbender occupancy. Although we found no difference in water quality parameters and streambed particle size among the stage classes at the sites of capture, there was a significant difference in the average shelter size among all stages, with larvae utilizing the smallest shelters. Based on these results, future Hellbender research and conservation efforts should consider differences in life stage habitat use as well as specific stream particle classes.

A new species of Gymnotus is described from coastal river systems in the Pernambuco State, Brazil. It is phylogenetically referred to the “Gymnotus carapo group clade” for presenting a clear patch posteriorly at anal fin, two independent pores at dorsoposterior corner of preopercle, a single row of well-developed teeth (most arrowhead-shaped) anteriorly on premaxilla, cleithrum with anterior notch, and by the relative anus to anal-fin distance, pectoral-fin length, and maxilla length. The new species is distinguished from all congeners in the “Gymnotus carapo group clade” by a unique set of characters of uncertain polarity, including the number of dark bands along the body, dark bands along body three to four times wider than pale interbands, with nearly straight margins (never broken anteriorly into irregular dark spots), and bands nearly uniform in color, number of scales above lateral line, number of lateral-line perforated scales to first ventral ramus, number of total pored lateral-line scales, head length, snout length, body depth, number of anal-fin rays, number of pectoral-fin rays, number of precaudal vertebrae, number of teeth on anterior row of premaxilla, and number of teeth along outer row of dentary. The new taxon represents the first species of Gymnotus described from localities in northeastern Brazil, north of the mouth of the rio São Francisco.