Sample records for ambystoma

  1. Ambystoma maculatum (spotted salamander). Reproduction

    Glorioso, Brad M.; Waddle, Hardin; Hefner, Jeromi


    The Spotted Salamander is a wide-ranging salamander of the eastern United States that typically breeds in winter or early spring in ephemeral pools in lowland forests. Ambystoma maculatum is known to deposit 2-4 egg masses per year, each containing 1-250 eggs. As part of ongoing research into the ecology and reproductive biology of Spotted Salamanders in the Kisatchie District of Kisatchie National Forest in Natchitoches Parish, Louisiana, USA, we have been counting the number of embryos per egg mass. We captured seven female A. maculatum in a small pool, six of which were still gravid. We took standard measurements, including SVL, and then implanted a Passive Integrated Transponder (PIT tag) into each adult female as was the protocol. About an hour after processing these animals we marked new A. maculatum egg masses found in the same small pool using PVC pin flags pushed carefully through the outer jelly. We did not have enough time to process them that evening, and it was not until a few days later that we photographed those masses. We discovered that one of the masses contained a PIT tag in the outer jelly that corresponded to one of the six gravid females that were marked that same evening. To our knowledge, this is the first report of PIT tags being the means, albeit coincidentally, by which a particular egg mass of Ambystoma maculatum has been assigned to a particular female. For our purposes, losing the PIT tag from the adult female is counter to the goals of our study of this population, and we will no longer be implanting PIT tags into gravid females.

  2. Final Critical Habitat for Reticulated Flatwoods Salamander (Ambystoma bishopi)

    US Fish and Wildlife Service, Department of the Interior — These data identify, in general, the areas of final critical habitat for the endangered Ambystoma bishopi (reticulated flatwoods salamander).

  3. Developmental dynamics of Ambystoma tigrinum in a changing landscape

    McMenamin Sarah K; Hadly Elizabeth A


    Abstract Background Loss of pond habitat is catastrophic to aquatic larval amphibians, but even reduction in the amount of time a breeding site holds water (hydroperiod) can influence amphibian development and limit reproductive success. Using the landscape variation of a glacial valley in the Greater Yellowstone Ecosystem as the context for a natural experiment, we examined variation in growth pattern and life history of the salamander Ambystoma tigrinum melanostictum and determined how thes...

  4. Habitat segregation of Ambystoma gracile and Ambystoma macrodactylum in mountain ponds, Mount Rainer National Park, Washington, USA

    Hoffman, Robert L.; Larson, Gary L.; Brokes, Brendan J.


    Ambystoma gracile (Baird) and Ambystoma macrodactylum Baird are common salamander species occupying key trophic positions in mountain ponds and lakes of Mount Rainier National Park (MORA). Based on amphibian surveys conducted in ponds and lakes in the northeast quadrant of MORA during the summers of 1993a??1995, 20 sites were resampled in 1996 to document and evaluate distributions of the two species relative to site habitat characteristics. Distributions of larvae were assessed either by nearshore snorkel or visual encounter surveys of each study site. Twelve environmental variables representing pond and lake physical characteristics, water quality, and nutrient concentrations were measured. The occurrences at each site of three major habitat components (i.e., pond bottom firmness, coarse woody debris, and emergent/aquatic vegetation) were qualitatively estimated. Allotopic distributions of larval populations of both species were related to site elevation, maximum depth, organic content of bottom substrates, and surface area. Ambystoma gracile sites were larger, deeper, lower in elevation, had flocculent sediment higher in organic content, abundant coarse woody debris, and limited emergent/aquatic vegetation relative to A. macrodactylum sites. Ambystoma macrodactylum sites were smaller, shallower, higher in elevation, had firm sediments low in organic content, and had a greater occurrence of emergent/aquatic vegetation than did A. gracile sites. Two sites supported syntopic populations and exhibited many of the habitat characteristics observed at sites inhabited by each species. The distribution of each species in MORA was related to the interconnection between habitat characteristics of ponds and lakes and species life-history patterns and possibly interspecific interactions.

  5. Temperature and ontogenetic effects on color change in the larval salamander species Ambystoma barbouri and Ambystoma texanum

    Temperature has been shown to affect body color in several species of amphibians. The interaction between color and temperature may also change over larval ontogeny, perhaps because of age-related or seasonal changes in selection pressures on color. We quantified the effects of temperature on the color of the salamander sister species Ambystoma barbouri and Ambystoma texanum over larval ontogeny. We found that early-stage larvae responded to cold temperatures with a dark color relative to that of the warm temperature response. Both species then exhibited an ontogenetic shift in larval color, with larvae becoming lighter with age. Interestingly, older larvae showed decreased plasticity in color change to temperature when compared with younger stages. Older A. texanum larvae exhibited a reversal in the direction of color change, with cold temperatures inducing a lighter color relative to warm temperatures. We suggest that the overall pattern of color change (a plastic color response to temperature for young larvae, a progressive lightening of larvae over development, and an apparent loss of color plasticity to temperature over ontogeny) can be plausibly explained by seasonal changes in environmental factors (temperature, ultraviolet radiation) selecting for body color. (author)

  6. Histological image data of limb skeletal tissue from larval and adult Ambystoma mexicanum.

    McCusker, Catherine D; Diaz-Castillo, Carlos; Sosnik, Julian; Phan, Anne; Gardiner, David M


    The data presented in this article are related to the article entitled "Cartilage and bone cells do not participate in skeletal regeneration in Ambystoma mexicanum limbs" [1]. Here we present image data of the post-embryonic development of the forelimb skeletal tissue of Ambystoma Mexicanum. Histological staining was performed on sections from the intact limbs of young (6.5 cm) and old (25 cm) animals, and on dissected skeletal tissues (cartilage, bone, and periosteum) from these animals. PMID:27547798

  7. Developmental dynamics of Ambystoma tigrinum in a changing landscape

    McMenamin Sarah K


    Full Text Available Abstract Background Loss of pond habitat is catastrophic to aquatic larval amphibians, but even reduction in the amount of time a breeding site holds water (hydroperiod can influence amphibian development and limit reproductive success. Using the landscape variation of a glacial valley in the Greater Yellowstone Ecosystem as the context for a natural experiment, we examined variation in growth pattern and life history of the salamander Ambystoma tigrinum melanostictum and determined how these developmental characteristics varied with hydroperiod over several summers. Results In ponds that dried early in the season, maximum larval size was reduced relative to the sizes achieved in permanent ponds. Ephemeral ponds were associated with early metamorphosis at small body sizes, while permanent ponds facilitated longer larval periods and later metamorphosis. Paedomorphosis resulted from indefinite metamorphic postponement, and was identified only in the most permanent environments. Patterns of growth and allometry were similar between ponds with different hydroperiods, but considerable life history variation was derived from modulating the timing of and size at metamorphosis. Considering maximum rates of growth and inferring the minimum size at metamorphosis across 25 ponds over the course of three years, we calculated that hydroperiods longer than three months are necessary to support these populations through metamorphosis and/or reproductive maturity. Conclusions Landscape heterogeneity fosters life history variation in this natural population. Modulation of the complex ambystomatid life cycle allows this species to survive in unpredictable environments, but current trends towards rapid pond drying will promote metamorphosis at smaller sizes and could eliminate the paedomorphic phenotype from this region. Metamorphosis at small size is has been linked to altered fitness traits, including reduced survival and fecundity. Thus, widespread

  8. Immunocytochemical localization and immunochemical characterization of an insulin-related peptide in the pancreas of the urodele amphibian, Ambystoma mexicanum

    Hansen, G N; Hansen, B L; Jørgensen, P N;


    sub-regions of the insulin molecule. Additionally, fractions of an ethanol-HCl extract of pancreatic tissue from Ambystoma was studied in both conventional dot-blot tests by means of the same panel of antibodies and a two-site sandwich time-resolved immunofluorometric assay for human insulin involving...... residues A8-10 and B26-30. In conclusion, we interpret the findings as indicating that the immunocytochemically revealed tissue bound antigen in the Ambystoma pancreatic B-cells may be a peptide related to human insulin....

  9. Diet of larval Ambystoma rivulare (Caudata: Ambystomatidae), a threatened salamander from the Volcán Nevado de Toluca, Mexico

    JULIO A LEMOS-ESPINAL; Smith, Geoffrey R.; Guillermo A. Woolrich-Piña; Raymundo Montoya-Ayala


    Several species of salamander in the genus Ambystoma occur in the mountains surrounding Mexico City and are considered at risk of extinction. However, little is known about their ecology and natural history. The Toluca Stream Siredon (Ambystoma rivulare) is classified as “Data Deficient” by the IUCN, and considered “Threatened” under Mexican law. From October 2013 to September 2014, we examined the diet of larval A. rivulare from a stream on the Volcán Nevado de Toluca in Mexico to provide in...

  10. Temporal response of the tiger salamander (Ambystoma tigrinum to 3,000 years of climatic variation

    Long Webb


    Full Text Available Abstract Background Amphibians are sensitive indicators of environmental conditions and show measurable responses, such as changes in phenology, abundance and range limits to local changes in precipitation and temperature regimes. Amphibians offer unique opportunities to study the important ecological and evolutionary implications of responses in life history characteristics to climatic change. We analyzed a late-Holocene fossil record of the Tiger Salamander (Ambystoma tigrinum for evidence of population-level changes in body size and paedomorphosis to climatic change over the last 3000 years. Results We found a significant difference in body size index between paedomorphic and metamorphic individuals during the time interval dominated by the Medieval Warm Period. There is a consistent ratio of paedomorphic to metamorphic specimens through the entire 3000 years, demonstrating that not all life history characteristics of the population were significantly altered by changes in climate on this timescale. Conclusion The fossil record of Ambystoma tigrinum we used spans an ecologically relevant timescale appropriate for understanding population and community response to projected climatic change. The population-level responses we documented are concordant with expectations based on modern environmental studies, and yield insight into population-level patterns across hundreds of generations, especially the independence of different life history characteristics. These conclusions lead us to offer general predictions about the future response of this species based on likely scenarios of climatic warming in the Rocky Mountain region.

  11. The Axolotl (Ambystoma mexicanum: Factors That Limit its Production and Alternatives for its Conservation

    J. Toca-Ramirez


    Full Text Available Ambystoma mexicanum is an amphibian endemic to Xochimilco Lake in Mexico City. It has been declared in danger of extinction and is under special protection. Some chemical contaminants in the water are extremely high and could be the cause of its high mortality rate in certain areas of Xochimilco. In order to preserve this species it will not only be necessary and fundamental to prohibit fishing axolotls in their natural state, a market study and nutritional chemical analysis will also be necessary in order to establish the organoleptic properties and level of acceptance before a taste panel; that is to say, get to know more about the specie in order to give the product added value offering its meat as an unconventional delicacy. This way the creation of farms that will help its conservation will be justified. On the other hand it is important to mention that the axolotls are very important in scientific research. Since it serves as an amphibious model for many physiological and morphological processes that explain the regenerative process that this species possess. The objective of this study is to emphasize the advantages that the Ambystoma mexicanum has with the intention to rationally exploit these attributes in order to achieve its conservation.

  12. Cryptic sex? Estimates of genome exchange in unisexual mole salamanders (Ambystoma sp.).

    Gibbs, H Lisle; Denton, Robert D


    Cryptic sex has been argued to explain the exceptional longevity of certain parthenogenetic vertebrate lineages, yet direct measurements of genetic exchange between sexual and apparently parthenogenetic forms are rare. Female unisexual mole salamanders (Ambystoma sp.) are the oldest known unisexual vertebrate lineage (~5 million years), and one hypothesis for their persistence is that allopolyploid female unisexuals periodically exchange haploid genomes 'genome exchange' during gynogenetic reproduction with males from sympatric sexual species. We test this hypothesis by using genome-specific microsatellite DNA markers to estimate the rates of genome exchange between sexual males and unisexual females in two ponds in NE Ohio. We also test the prediction that levels of gene flow should be higher for 'sympatric' (sexual males present) genomes in unisexuals compared to 'allopatric' (sexual males absent) unisexual genomes. We used a model testing framework in the coalescent-based program MIGRATE-N to compare models where unidirectional gene flow is present and absent between sexual species and unisexuals. As predicted, our results show higher levels of gene flow between sexuals and sympatric unisexual genomes compared to lower (likely artefactual) levels of gene flow between sexuals and allopatric unisexual genomes. Our results provide direct evidence that genome exchange between sexual and unisexual Ambystoma occurs and demonstrate that the magnitude depends on which sexual species are present. The relatively high levels of gene flow suggest that unisexuals must be at a selective advantage over sexual forms so as to avoid extinction due to genetic swamping through genome exchange. PMID:27100619

  13. Maintenance media for the axolotl Ambystoma mexicanum juveniles (Amphibia: Caudata Soluciones de mantenimiento de juveniles del ajolote Ambystoma mexicanum (Amphibia: Caudata

    Cecilia Robles Mendoza


    Full Text Available Physiological condition and organisms' health which are grown in culture systems depends on several factors including water quality, feeding and density among others. In Mexico, the colonies of the axolotl Ambystoma mexicanum (Shaw, an indigenous amphibian under extinction risk, are maintained under different culture conditions according to the objectives of the colony and the available resources. Particularly, water electrolytic characteristic and ionic and osmotic conditions are the factors with greater variation in the axolotl culture systems. Therefore, it is necessary to standardize the best maintenance conditions to store the germoplasm of the axolotl and to ensure healthy organisms with researching purposes. Thus, the aim of the present study was to evaluate the development and growth of Ambystoma mexicanum larvae reared under different maintenance media, usually used in Mexico for the culture of the species: 1 dechlorinated tap water; 2 dechlorinated tap water enriched with sodium chloride and commercial colloidal solution and 3 Holtfreter's solution reconstructed with dechlorinated tap water. In each experimental condition, 15 larvae on stage 44 (immediately after hatching were maintained during 21 days and development and growth were weekly recorded. Ionic and osmotic conditions of the external media were routinely registered. The obtained results suggested a better physiological condition of the axolotls maintained on Holtfreter's solution, where the highest growth rate (13 g WW d-1 and the greatest condition factor (0.79 were registered. The use of this solution is recommended due it guarantees the suitable development of early stages of A. mexicanum on culture systems.La condición fisiológica y por lo tanto la salud de los organismos acuáticos depende de varios factores como la calidad del agua de mantenimiento, la alimentación, la densidad, entre otros. En México, las colonias del ajolote Ambystoma mexicanum (Shaw, anfibio end

  14. Identification of essential and non-essential genes in Ambystoma tigrinum virus.

    Aron, Mariah M; Allen, Alexander G; Kromer, Mathew; Galvez, Hector; Vigil, Brianna; Jancovich, James K


    Members of the genus Ranavirus (family Iridoviridae) are large double-stranded (ds) DNA viruses that are found world-wide infecting fish, amphibian and reptile ectothermic hosts. Ranavirus genomes range from 105 to 155kbp in length and they are predicted to encode around 90-125 genes. Currently, our knowledge of the function of ∼50% of these genes is known or inferred based on homology to orthologous genes characterized in other systems; however, the function of the remaining open reading frames (ORFS) is unknown. Therefore, in order to begin to uncover the function of unknown ORFs in ranaviruses we developed a standardized approach to generate a recombination cassette for any ORF in Ambystoma tigrinum virus (ATV). Our standardized approach quickly and efficiently assembles recombination cassettes and recombinant ATV. We have used this approach to identify two essential, one semi-essential and two non-essential genes in ATV. PMID:27025572

  15. Effect of water quality on the feeding ecology of axolotl Ambystoma mexicanum

    Diego de Jesus Chaparro-Herrera


    Full Text Available Ambystoma mexicanum, a highly endangered species, is endemic to lake Xochimilco (Mexico City, Mexico which currently is being negatively affected by the introduction of Oreochromis niloticus (Tilapia and water pollution. During the first weeks of development, when mortality is the highest, Ambystoma mexicanumdepends on a diet of zooplankton. The aim of this study was to check whether contamination levels in lake Xochimilco influence zooplankton consumption by similar size classes of A. mexicanum and Oreochromis niloticus. In this study, we analysed changes in the functional responses and prey preference of A. mexicanum and larval Tilapia in two media, one with filtered lake Xochimilco water and another one with reconstituted water. As prey we used cladocerans (Moina macrocopa, Alona glabra, Macrothrix triserialis and Simocephalus vetulus and ostracods (Heterocypris incongruens. Zooplankton was offered in 5 different densities, 10, 20, 40, 80, 160 ind./mL. Prey consumption by A. mexicanum varied in relation to the species offered and age of the larvae. From the first week to the eighth week prey consumption by A. mexicanum increased by 57%. Our functional response tests showed that regardless of the prey type, prey consumption by A. mexicanum was lower in the contaminated water from lake Xochimilco. Among the zooplankton offered in the contaminated environment predators preferred smaller and slower moving microcrustaceans such as Alona glabra and Heterocypris incongruens. Furthermore, O. niloticus preferred prey such as Moina macrocopa and Macrothrix triserialis in the contaminated medium and was more voracious than the axolotl. Our results indicate that both water quality of the lake and the presence of the more resistant exotic fish adversely impact the survival of this endangered amphibian.

  16. Evaluation of the anesthetic effects of MS222 in the adult Mexican axolotl (Ambystoma mexicanum

    Zullian C


    Full Text Available Chiara Zullian,1 Aurore Dodelet-Devillers,1 Stéphane Roy,2 Pascal Vachon1 1Département de Biomédecine Vétérinaire, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, 2Département de Stomatologie, Faculté de Médecine Dentaire, Montréal, Québec, Canada Abstract: The Mexican axolotl (Ambystoma mexicanum is a unique research model in several fields of medicine, where surgical and invasive procedures may be required. As yet, little is known about the efficacy of MS222 (tricaine methanesulfonate, which is the most commonly used anesthetic agent in amphibians. The main objectives of this study were to evaluate the anesthetic effects and physiological changes in adult axolotls following a 20-minute immersion bath, containing progressive MS222 concentrations starting at 0.1%. Depth of anesthesia and physiological changes were evaluated every 15 minutes post-MS222 exposure with the following parameters: righting behavior, withdrawal reflex, acetic acid test response, heart rate, and blood oxygen saturation, as well as cloacal and body surface temperatures. A 20-minute exposure in a 0.1% MS222 immersion bath (n=6 animals had no anesthetic effects on adult axolotls after 20 minutes of exposure. With a 0.2% MS222 solution, all axolotls (n=9 were deeply anesthetized at 15 minutes, and 80% were still unresponsive at 30 minutes postexposure. Blood oxygen saturation and heart rate were slightly, but significantly, increased when compared with the baseline value and remained stable up to recovery. There was no significant increase in surface and cloaca temperatures, compared with baseline. With the 0.4% MS222 solution, the duration of anesthesia lasted for 90 minutes to at least 120 minutes (n=3 animals and this concentration was deemed too high. In conclusion, a 20-minute immersion bath with 0.2% MS222 may be used for short procedures (15–30 minutes requiring anesthesia of adult axolotls. Keywords: Ambystoma mexicanum

  17. Proteinaceous Pheromone Homologs Identified from the Cloacal Gland Transcriptome of a Male Axolotl, Ambystoma mexicanum.

    Kevin W Hall

    Full Text Available Pheromones play an important role in modifying vertebrate behavior, especially during courtship and mating. Courtship behavior in urodele amphibians often includes female exposure to secretions from the cloacal gland, as well as other scent glands. The first vertebrate proteinaceous pheromone discovered, the decapeptide sodefrin, is a female attracting pheromone secreted by the cloacal gland of male Cynops pyrrhogaster. Other proteinaceous pheromones in salamanders have been shown to elicit responses from females towards conspecific males. The presence and levels of expression of proteinaceous pheromones have not been identified in the family Ambystomatidae, which includes several important research models. The objective of this research was therefore to identify putative proteinaceous pheromones from male axolotls, Ambystoma mexicanum, as well as their relative expression levels. The results indicate that axolotls possess two different forms of sodefrin precursor-like factor (alpha and beta, as well as a putative ortholog of plethodontid modulating factor. The beta form of sodefrin precursor-like factor was amongst the most highly expressed transcripts within the cloacal gland. The ortholog of plethodontid modulating factor was expressed at a level equivalent to the beta sodefrin precursor-like factor. The results are from a single male axolotl; therefore, we are unable to assess how representative our results may be. Nevertheless, the presence of these highly expressed proteinaceous pheromones suggests that male axolotls use multiple chemical cues to attract female conspecifics. Behavioral assays would indicate whether the putative protein pheromones elicit courtship activity from female axolotls.

  18. Effects of copper exposure on hatching success and early larval survival in marbled salamanders, Ambystoma opacum.

    Soteropoulos, Diana L; Lance, Stacey L; Flynn, R Wesley; Scott, David E


    The creation of wetlands, such as urban and industrial ponds, has increased in recent decades, and these wetlands often become enriched in pollutants over time. One metal contaminant trapped in created wetlands is copper (Cu(2+)). Copper concentrations in sediments and overlying water may affect amphibian species that breed in created wetlands. The authors analyzed the Cu concentration in dried sediments from a contaminated wetland and the levels of aqueous Cu released after flooding the sediments with different volumes of water, mimicking low, medium, and high pond-filling events. Eggs and larvae of Ambystoma opacum Gravenhorst, a salamander that lays eggs on the sediments in dry pond beds that hatch on pond-filling, were exposed to a range of Cu concentrations that bracketed potential aqueous Cu levels in created wetlands. Embryo survival varied among clutches, but increased Cu levels did not affect embryo survival. At Cu concentrations of 500 µg/L or greater, however, embryos hatched earlier, and the aquatic larvae died shortly after hatching. Because Cu concentrations in sediments increase over time in created wetlands, even relatively tolerant species such as A. opacum may be affected by Cu levels in the posthatching environment. PMID:24729474

  19. Neural crest does not contribute to the neck and shoulder in the axolotl (Ambystoma mexicanum.

    Hans-Henning Epperlein

    Full Text Available BACKGROUND: A major step during the evolution of tetrapods was their transition from water to land. This process involved the reduction or complete loss of the dermal bones that made up connections to the skull and a concomitant enlargement of the endochondral shoulder girdle. In the mouse the latter is derived from three separate embryonic sources: lateral plate mesoderm, somites, and neural crest. The neural crest was suggested to sustain the muscle attachments. How this complex composition of the endochondral shoulder girdle arose during evolution and whether it is shared by all tetrapods is unknown. Salamanders that lack dermal bone within their shoulder girdle were of special interest for a possible contribution of the neural crest to the endochondral elements and muscle attachment sites, and we therefore studied them in this context. RESULTS: We grafted neural crest from GFP+ fluorescent transgenic axolotl (Ambystoma mexicanum donor embryos into white (d/d axolotl hosts and followed the presence of neural crest cells within the cartilage of the shoulder girdle and the connective tissue of muscle attachment sites of the neck-shoulder region. Strikingly, neural crest cells did not contribute to any part of the endochondral shoulder girdle or to the connective tissue at muscle attachment sites in axolotl. CONCLUSIONS: Our results in axolotl suggest that neural crest does not serve a general function in vertebrate shoulder muscle attachment sites as predicted by the "muscle scaffold theory," and that it is not necessary to maintain connectivity of the endochondral shoulder girdle to the skull. Our data support the possibility that the contribution of the neural crest to the endochondral shoulder girdle, which is observed in the mouse, arose de novo in mammals as a developmental basis for their skeletal synapomorphies. This further supports the hypothesis of an increased neural crest diversification during vertebrate evolution.

  20. Impacts of a gape limited Brook Trout, Salvelinus fontinalis, on larval Northwestern salamander, Ambystoma gracile, growth: A field enclosure experiment

    Currens, C.R.; Liss, W.J.; Hoffman, R.L.


    The formation of amphibian population structure is directly affected by predation. Although aquatic predators have been shown to have direct negative effects on larval salamanders in laboratory and field experiments, the potential impacts of gape-limited fish on larval salamander growth has been largely underexplored. We designed an enclosure experiment conducted in situ to quantify the effects of gape-limited Brook Trout (Salvelinus fontinalis) on larval Northwestern Salamander (Ambystoma gracile) growth. We specifically tested whether the presence of fish too small to consume larvae had a negative effect on larval growth. The results of this study indicate that the presence of a gape-limited S. fontinalis can have a negative effect on growth of larval A. gracile salamanders. Copyright 2007 Society for the Study of Amphibians and Reptiles.

  1. Metals, Parasites, and Environmental Conditions Affecting Breeding Populations of Spotted Salamanders (Ambystoma maculatum) in Northern Arkansas, USA.

    DeMali, Heather M; Trauth, Stanley E; Bouldin, Jennifer L


    The spotted salamander (Ambystoma maculatum) is indigenous to northern Arkansas, and several breeding sites are known to exist in the region. Spotted salamanders (n = 17) were collected and examined for parasites and only three females harbored nematodes (Physaloptera spp.). Chronic aquatic bioassays were conducted using water collected from eight breeding ponds during different hydroperiod events. No lethal or sublethal effects were measured in Ceriodaphnia dubia; however, decreased growth and survival were seen in Pimephales promelas. Aqueous, sediment, and salamander hepatic samples were analyzed for As, Cd, Cu, Pb, and Ni. Metal analysis revealed possible increased metal exposure following precipitation, with greatest metal concentrations measured in sediment samples. Hepatic metal concentrations were similar in parasitized and non-parasitized individuals, and greatest Pb concentrations were measured following normal precipitation events. Determining environmental stressors of amphibians, especially during their breeding and subsequent larval life stage, is imperative to improve species conservation. PMID:26886425

  2. Amphibian Sex Determination: Segregation and Linkage Analysis Using Members of the Tiger Salamander Species Complex (Ambystoma mexicanum and A. t. tigrinum)

    Smith, Jeramiah J.; Voss, S. Randal


    Little is known about the genetic basis of sex determination in vertebrates though considerable progress has been made in recent years. In this study, segregation analysis and linkage mapping were performed to localize an amphibian sex-determining locus (ambysex) in the tiger salamander (Ambystoma) genome. Segregation of sex phenotypes (male, female) among 2nd generation individuals of interspecific crosses (A. mexicanum x A. t. tigrinum) was consistent with Mendelian expectations, although a...

  3. Breeding biology of the spotted salamander Ambystoma maculatum (Shaw) in acidic temporary ponds at Cape Cod, USA

    Portnoy, J.W.


    The relationship between water chemistry and breeding success of spotted salamanders Ambystoma maculatum (Shaw) was examined in temporary woodland ponds on outer Cape Cod, Massachusetts in 1985 and 1986. Most pond waters were dilute (3median coductivity = 57 umhos cm-1 (1 umhos cm-1 = 0?1 mSm-1)), acidic (median pH = 4?82), and highly colored (median = 140 Pt-Co units). Most acidity was due to abundant organic acids. Salamander survival to hatching was over 80% at 8 of 12 ponds monitored. Complete mortality, preceded by gross abnormalities, was observed only among embryos in the most acidic spawning pond (pH 4?3-4?5) in both years. Embryo transfers between ponds and laboratory studies indicated that reduced survival was due to the interaction of low pH with high tannin-lignin concentration. The use of amphibian embryonic survival to indicate acid rain effects is complicated by multiple habitat parameters and should only be attempted in conjunction with long-term population monitoring.

  4. The effects of simulated solar UVB radiation on early developmental stages of the Northwestern Salamander (Ambystoma gracile) from three lakes

    Calfee, Robin D.; Little, Edward E.; Pearl, Christopher A.; Hoffman, Robert L.


    Solar ultraviolet radiation (UV) has received much attention as a factor that could play a role in amphibian population declines. UV can be hazardous to some amphibians, but the resultant effects depend on a variety of environmental and behavioral factors. In this study, the potential effects of UV on the Northwestern Salamander, Ambystoma gracile, from three lakes were assessed in the laboratory using a solar simulator. We measured the survival of embryos and the survival and growth of larvae exposed to four UV treatments in controlled laboratory studies, the UV absorbance of egg jelly, oviposition depths in the lakes, and UV absorbance in water samples from the three lakes. Hatching success of embryos decreased in the higher UV treatments as compared to the control treatments, and growth of surviving larvae was significantly reduced in the higher UVB irradiance treatments. The egg jelly exhibited a small peak of absorbance within the UVB range (290–320 nm). The magnitude of UV absorbance differed among egg jellies from the three lakes. Oviposition depths at the three sites averaged 1.10 m below the water surface. Approximately 66% of surface UVB radiation was attenuated at 10-cm depth in all three lakes. Results of this study indicate that larvae may be sensitive to UVB exposure under laboratory conditions; however, in field conditions the depths of egg deposition in the lakes, absorbance of UV radiation by the water column, and the potential for behavioral adjustments may mitigate severe effects of UV radiation.

  5. Effect of thyroid hormone concentration on the transcriptional response underlying induced metamorphosis in the Mexican axolotl (Ambystoma

    Samuels Amy K


    Full Text Available Abstract Background Thyroid hormones (TH induce gene expression programs that orchestrate amphibian metamorphosis. In contrast to anurans, many salamanders do not undergo metamorphosis in nature. However, they can be induced to undergo metamorphosis via exposure to thyroxine (T4. We induced metamorphosis in juvenile Mexican axolotls (Ambystoma mexicanum using 5 and 50 nM T4, collected epidermal tissue from the head at four time points (Days 0, 2, 12, 28, and used microarray analysis to quantify mRNA abundances. Results Individuals reared in the higher T4 concentration initiated morphological and transcriptional changes earlier and completed metamorphosis by Day 28. In contrast, initiation of metamorphosis was delayed in the lower T4 concentration and none of the individuals completed metamorphosis by Day 28. We identified 402 genes that were statistically differentially expressed by ≥ two-fold between T4 treatments at one or more non-Day 0 sampling times. To complement this analysis, we used linear and quadratic regression to identify 542 and 709 genes that were differentially expressed by ≥ two-fold in the 5 and 50 nM T4 treatments, respectively. Conclusion We found that T4 concentration affected the timing of gene expression and the shape of temporal gene expression profiles. However, essentially all of the identified genes were similarly affected by 5 and 50 nM T4. We discuss genes and biological processes that appear to be common to salamander and anuran metamorphosis, and also highlight clear transcriptional differences. Our results show that gene expression in axolotls is diverse and precise, and that axolotls provide new insights about amphibian metamorphosis.

  6. Effects of simulated solar UVB radiation on early developmental stages of the northwestern salamander (Ambystoma gracile) from three lakes

    Calfee, R.D.; Little, E.E.; Pearl, C.A.; Hoffman, R.L.


    Solar ultraviolet radiation (UV) has received much attention as a factor that could play a role in amphibian population declines. UV can be hazardous to some amphibians, but the resultant effects depend on a variety of environmental and behavioral factors. In this study, the potential effects of UV on the Northwestern Salamander, Ambystoma gracile, from three lakes were assessed in the laboratory using a solar simulator. We measured the survival of embryos and the survival and growth of larvae exposed to four UV treatments in controlled laboratory studies, the UV absorbance of egg jelly, oviposition depths in the lakes, and UV absorbance in water samples from the three lakes. Hatching success of embryos decreased in the higher UV treatments as compared to the control treatments, and growth of surviving larvae was significantly reduced in the higher UVB irradiance treatments. The egg jelly exhibited a small peak of absorbance within the UVB range (290-320 nm). The magnitude of UV absorbance differed among egg jellies from the three lakes. Oviposition depths at the three sites averaged 1.10 m below the water surface. Approximately 66 of surface UVB radiation was attenuated at 10-cm depth in all three lakes. Results of this study indicate that larvae may be sensitive to UVB exposure under laboratory conditions; however, in field conditions the depths of egg deposition in the lakes, absorbance of UV radiation by the water column, and the potential for behavioral adjustments may mitigate severe effects of UV radiation. Copyright 2010 Society for the Study of Amphibians and Reptiles.

  7. Effects of metamorphosis and captivity on the in vitro sensitivity of thyroid glands from the tiger salamander, Ambystoma tigrinum, to bovine thyrotropin

    The sensitivity of thyroid glands from the tiger salamander, Ambystoma tigrinum, to bovine thyrotropin (bTSH) was tested in vitro. Thyroids were taken from subjects representing metamorphic stages I (premetamorphic larvae), II (onset of climax), and VII (completion of gill resorption), as well as from captivity control larvae. Exogenous TSH reduced the cumulative uptake of 125I in vitro by thyroids from stage I larvae after 24 and 48 hr. The capacity of thyroids to release thyroxine (T4) in vitro was used subsequently as a measure of their responsiveness to TSH. Baseline levels of T4 release in vitro were variable but did not differ significantly among developmental stages. A low dose of bTSH (5 X 10(-6) IU/ml) did not increase in vitro T4 release compared with that of controls. A larger dose (5 X 10(-4) IU/ml) caused greater increases in T4 release from thyroids of stage II and VII subjects than from those of controls. This dose produced only a small response by thyroids from captivity-control subjects. The results suggest that the thyroids of Ambystoma increase in their capacity to respond to TSH during the process of metamorphosis

  8. Regeneración miocárdica en Ambystoma mexicanum después de lesión quirúrgica

    Vargas-González,Alvaro; Prado-Zayago,Esteban; León-Olea, Martha; Guarner-Lans, Verónica; Cano-Martínez, Agustina


    Se realizó resección ventricular en el corazón de Ambystoma mexicanum, se evaluó si la restitución del tejido resulta de hipertrofia o de hiperplasia. Por medio de una tinción tricrómica se encontró que 5 días después del daño en el espacio de la resección se encontró un coágulo rodeado de fibras de colágena (83 ± 6%), músculo (10 ± 3%) y zonas sin tejido (7 ± 2%). Una proporción de 50 ± 4 y 90 ± 2% correspondió a tejido muscular 10 y 30 días después de la lesión. La tinción con bis-Benzimida...

  9. A populational survey of 45S rDNA polymorphism in the Jefferson salamander Ambystoma jeffersonianum revealed by fluorescence in situ hybridization (FISH)

    Ke BI; James P.BOGART; Jinzhong FU


    The chromosomal localization of 45S ribosomal RNA genes in Ambystoma jeffersonianum was determined by fluorescence in situ hybridization with 18S rDNA fragment as a probe (FISH-rDNA). Our results revealed the presence of rDNA polymorphism among A.jeffersonianum populations in terms of number, location and FISH signal intensity on the chromosomes. Nine rDNA cytotypes were found in ten geographically isolated populations and most of them contained derivative rDNA sites. Our preliminary study provides strong indication of karyotypic diversification of A.jeffersonianum that is demonstrated by intraspecific variation of 45S rDNA cytotypes. rDNA cytotype polymorphism has been described in many other caudate amphibians. We predict that habitat isolation, low dispersal ability and decline of effective population size could facilitate the fixation and accumulation of variable rDNA cytotypes during their chromosome evolution [Current Zoology 55(2):145-149,2009].

  10. Role of habitat complexity in predator-prey dynamics between an introduced fish and larval Long-toed Salamanders (Ambystoma macrodactylum)

    Kenison, Erin K; Litt, Andrea R.; Pilliod, David; McMahon, Tom E


    Predation by nonnative fishes has reduced abundance and increased extinction risk for amphibian populations worldwide. Although rare, fish and palatable amphibians have been observed to coexist where aquatic vegetation and structural complexity provide suitable refugia. We examined whether larval long-toed salamanders (Ambystoma macrodactylum Baird, 1849) increased use of vegetation cover in lakes with trout and whether adding vegetation structure could reduce predation risk and nonconsumptive effects (NCEs), such as reductions in body size and delayed metamorphosis. We compared use of vegetation cover by larval salamanders in lakes with and without trout and conducted a field experiment to investigate the influence of added vegetation structure on salamander body morphology and life history. The probability of catching salamanders in traps in lakes with trout was positively correlated with the proportion of submerged vegetation and surface cover. Growth rates of salamanders in enclosures with trout cues decreased as much as 85% and the probability of metamorphosis decreased by 56%. We did not find evidence that adding vegetation reduced NCEs in experimental enclosures, but salamanders in lakes with trout utilized more highly-vegetated areas which suggests that adding vegetation structure at the scale of the whole lake may facilitate coexistence between salamanders and introduced trout.

  11. Microhabitat types promote the genetic structure of a micro-endemic and critically endangered mole salamander (Ambystoma leorae) of Central Mexico.

    Sunny, Armando; Monroy-Vilchis, Octavio; Reyna-Valencia, Carlos; Zarco-González, Martha M


    The reduced immigration and emigration rates resulting from the lack of landscape connectivity of patches and the hospitality of the intervening matrix could favor the loss of alleles through genetic drift and an increased chance of inbreeding. In order for isolated populations to maintain sufficient levels of genetic diversity and adapt to environmental changes, one important conservation goal must be to preserve or reestablish connectivity among patches in a fragmented landscape. We studied the last known population of Ambystoma leorae, an endemic and critically threatened species. The aims of this study were: (1) to assess the demographic parameters of A. leorae and to distinguish and characterize the microhabitats in the river, (2) to determine the number of existing genetic groups or demes of A. leorae and to describe possible relationships between microhabitats types and demes, (3) to determine gene flow between demes, and (4) to search for geographic locations of genetic discontinuities that limit gene flow between demes. We found three types of microhabitats and three genetically differentiated subpopulations with a significant level of genetic structure. In addition, we found slight genetic barriers. Our results suggest that mole salamander's species are very sensitive to microhabitat features and relatively narrow obstacles in their path. The estimates of bidirectional gene flow are consistent with the pattern of a stepping stone model between demes, where migration occurs between adjacent demes, but there is low gene flow between distant demes. We can also conclude that there is a positive correlation between microhabitats and genetic structure in this population. PMID:25076052

  12. Caracterización de la Infección Parasitaria en Ambystoma andersoni Krebs & Brandon‚ 1984 en la Laguna de Zacapu‚ Michoacán‚ México -resumen-

    L O Alvarez-Silva; Y Herrerías; D Huacuz-Elías; M A Alvarez-Ramírez


    Ambystoma andersoni especie neoténica y endémica a la Laguna de Zacapu‚ ubicada dentro del municipio con el mismo nombre‚ en el norte de estado de Michoacán‚ lo que la hace particularmente especial dado que se encuentra dentro de los límites de distribución del género. Existen pocos estudios acerca de está‚ sin embargo la evidencia empírica sugiere que las especies de anfibios son susceptibles a cambios en el medio‚ los cuales pueden promover diversas enfermedades que alteran los atributos pr...

  13. Evolution of vertebrate IgM: complete amino acid sequence of the constant region of Ambystoma mexicanum mu chain deduced from cDNA sequence.

    Fellah, J S; Wiles, M V; Charlemagne, J; Schwager, J


    cDNA clones coding for the constant region of the Mexican axolotl (Ambystoma mexicanum) mu heavy immunoglobulin chain were selected from total spleen RNA, using a cDNA polymerase chain reaction technique. The specific 5'-end primer was an oligonucleotide homologous to the JH segment of Xenopus laevis mu chain. One of the clones, JHA/3, corresponded to the complete constant region of the axolotl mu chain, consisting of a 1362-nucleotide sequence coding for a polypeptide of 454 amino acids followed in 3' direction by a 179-nucleotide untranslated region and a polyA+ tail. The axolotl C mu is divided into four typical domains (C mu 1-C mu 4) and can be aligned with the Xenopus C mu with an overall identity of 56% at the nucleotide level. Percent identities were particularly high between C mu 1 (59%) and C mu 4 (71%). The C-terminal 20-amino acid segment which constitutes the secretory part of the mu chain is strongly homologous to the equivalent sequences of chondrichthyans and of other tetrapods, including a conserved N-linked oligosaccharide, the penultimate cysteine and the C-terminal lysine. The four C mu domains of 13 vertebrate species ranging from chondrichthyans to mammals were aligned and compared at the amino acid level. The significant number of mu-specific residues which are conserved into each of the four C mu domains argues for a continuous line of evolution of the vertebrate mu chain. This notion was confirmed by the ability to reconstitute a consistent vertebrate evolution tree based on the phylogenic parsimony analysis of the C mu 4 sequences. PMID:1382992

  14. Microhabitat types promote the genetic structure of a micro-endemic and critically endangered mole salamander (Ambystoma leorae of Central Mexico.

    Armando Sunny

    Full Text Available The reduced immigration and emigration rates resulting from the lack of landscape connectivity of patches and the hospitality of the intervening matrix could favor the loss of alleles through genetic drift and an increased chance of inbreeding. In order for isolated populations to maintain sufficient levels of genetic diversity and adapt to environmental changes, one important conservation goal must be to preserve or reestablish connectivity among patches in a fragmented landscape. We studied the last known population of Ambystoma leorae, an endemic and critically threatened species. The aims of this study were: (1 to assess the demographic parameters of A. leorae and to distinguish and characterize the microhabitats in the river, (2 to determine the number of existing genetic groups or demes of A. leorae and to describe possible relationships between microhabitats types and demes, (3 to determine gene flow between demes, and (4 to search for geographic locations of genetic discontinuities that limit gene flow between demes. We found three types of microhabitats and three genetically differentiated subpopulations with a significant level of genetic structure. In addition, we found slight genetic barriers. Our results suggest that mole salamander's species are very sensitive to microhabitat features and relatively narrow obstacles in their path. The estimates of bidirectional gene flow are consistent with the pattern of a stepping stone model between demes, where migration occurs between adjacent demes, but there is low gene flow between distant demes. We can also conclude that there is a positive correlation between microhabitats and genetic structure in this population.

  15. Caracterización de la Infección Parasitaria en Ambystoma andersoni Krebs & Brandon‚ 1984 en la Laguna de Zacapu‚ Michoacán‚ México -resumen-

    L O Alvarez-Silva


    Full Text Available Ambystoma andersoni especie neoténica y endémica a la Laguna de Zacapu‚ ubicada dentro del municipio con el mismo nombre‚ en el norte de estado de Michoacán‚ lo que la hace particularmente especial dado que se encuentra dentro de los límites de distribución del género. Existen pocos estudios acerca de está‚ sin embargo la evidencia empírica sugiere que las especies de anfibios son susceptibles a cambios en el medio‚ los cuales pueden promover diversas enfermedades que alteran los atributos primarios de las poblaciones‚ algunas de estas causadas por parásitos. Se realizó un muestreo de mayo del 2011 a abril del 2012‚ en cuatro sitios de la laguna para así tener una mejor representación. Se revisaron un total de 170 individuos‚ de los cuales el 58% se encuentran parasitados por algún tipo de parásito y el 42% está libre de infección. De los 98 individuos parasitados se determinaron infecciones monoespecíficas en el 57%‚ diespecíficas en el 30%‚ triespecíficas en el 12% y tetraespecífica con el 1%. Teniendo como resultados dos especies de ectoparásitos (Argulus ambystoma y Lernea sp.‚ cuatro especies de parásitos intestinales (Hedruris siredonis‚ Falcaustra chabaudi‚ Cosmocercoides dukae y Macroderoididae sp. y una especie hemoparásito (Trichodina sp.. Dentro de los ectoparásitos sólo el 32% del total estaba parasitado‚ con una prevalencia de 27,49% para Argulus ambystoma y 9,36% para Lernea sp. En cuanto a la abundancia promedio los resultados fueron: A. ambystoma 0,596 y Lernea sp 0,251. La intensidad promedio fue: A. ambystoma con 2,17 y Lernea sp con 2,67. Para los parásitos intestinales solo se revisaron 55 intestinos‚ de los cuales solo el 76% estaba parasitado. La prevalencia de parásitos intestinales fue: H. siredonis 13,45%; F. chabaudi 23,39%; C. dukae 2,34% y Macroderoididae sp 2,34%. En cuanto a la abundancia los resultados fueron: H. siredonis 0,614; F. chabaudi 8,719; C. dukae 0

  16. Regeneration of limb joints in the axolotl (Ambystoma mexicanum.

    Jangwoo Lee

    Full Text Available In spite of numerous investigations of regenerating salamander limbs, little attention has been paid to the details of how joints are reformed. An understanding of the process and mechanisms of joint regeneration in this model system for tetrapod limb regeneration would provide insights into developing novel therapies for inducing joint regeneration in humans. To this end, we have used the axolotl (Mexican Salamander model of limb regeneration to describe the morphology and the expression patterns of marker genes during joint regeneration in response to limb amputation. These data are consistent with the hypothesis that the mechanisms of joint formation whether it be development or regeneration are conserved. We also have determined that defects in the epiphyseal region of both forelimbs and hind limbs in the axolotl are regenerated only when the defect is small. As is the case with defects in the diaphysis, there is a critical size above which the endogenous regenerative response is not sufficient to regenerate the joint. This non-regenerative response in an animal that has the ability to regenerate perfectly provides the opportunity to screen for the signaling pathways to induce regeneration of articular cartilage and joints.

  17. Pattern formation in artificially activated ectoderm (Rana pipiens and Ambystoma punctatum)

    Nieuwkoop, P.D.


    Presumptive ectoneuroderm of late blastulae or early gastrulae of Rana pipiens was partially activated by short-lasting disaggregation in Ca-free Holtfreter or Niu-Twitty solutions and subsequent reaggregation in normal solutions. The explants usually became dumbbell shaped and consisted respectivel

  18. Road to evolution? Local adaptation to road adjacency in an amphibian (Ambystoma maculatum)

    Steven P. Brady


    The network of roads on the landscape is vast, and contributes a suite of negative ecological effects on adjacent habitats, ranging from fragmentation to contamination by runoff. In addition to the immediate consequences faced by biota living in roaded landscapes, road effects may further function as novel agents of selection, setting the stage for contemporary evolutionary changes in local populations. Though the ecological consequences of roads are well described, evolutionary outcomes rema...

  19. Suppression of first cleavage in the Mexican axolotl (Ambystoma mexicanum) by heat shock or hydrostatic pressure

    Gillespie, L.L.; Armstrong, J.B.


    Androgenetic diploid axolotls were produced by ultraviolet inactivation of the egg pronucleus shortly after fertilization, followed by suppression of the first cleavage division by hydrostatic pressure or heat shock. After treatment at 14,000 psi for 8 minutes, diploidy was restored in 74% of the embryos, but only 0.8% survived to hatching. A 36-37 degrees C heat shock of 10-minutes duration, applied 5.5 hours after the eggs were collected, yielded a slightly lower percentage of diploids. However, the proportion surviving to hatching was significantly greater (up to 4.6%). A second generation of androgenetic diploids was produced from one of the oldest of the first generation males with a similar degree of success. The lack of significant improvement suggests that the low survival is due to the heat shock per se and not to the uncovering of recessive lethal genes carried by the parent.

  20. Positional information is reprogrammed in blastema cells of the regenerating limb of the axolotl (Ambystoma mexicanum.

    Catherine D McCusker

    Full Text Available The regenerating region of an amputated salamander limb, known as the blastema, has the amazing capacity to replace exactly the missing structures. By grafting cells from different stages and regions of blastemas induced to form on donor animals expressing Green Fluorescent Protein (GFP, to non-GFP host animals, we have determined that the cells from early stage blastemas, as well as cells at the tip of late stage blastemas are developmentally labile such that their positional identity is reprogrammed by interactions with more proximal cells with stable positional information. In contrast, cells from the adjacent, more proximal stump tissues as well as the basal region of late bud blastemas are positionally stable, and thus form ectopic limb structures when grafted. Finally, we have found that a nerve is required to maintain the blastema cells in a positionally labile state, thus indicating a role for reprogramming cues in the blastema microenvironment.

  1. Regulation of Axolotl (Ambystoma mexicanum Limb Blastema Cell Proliferation by Nerves and BMP2 in Organotypic Slice Culture.

    Jeffrey Lehrberg

    Full Text Available We have modified and optimized the technique of organotypic slice culture in order to study the mechanisms regulating growth and pattern formation in regenerating axolotl limb blastemas. Blastema cells maintain many of the behaviors that are characteristic of blastemas in vivo when cultured as slices in vitro, including rates of proliferation that are comparable to what has been reported in vivo. Because the blastema slices can be cultured in basal medium without fetal bovine serum, it was possible to test the response of blastema cells to signaling molecules present in serum, as well as those produced by nerves. We also were able to investigate the response of blastema cells to experimentally regulated changes in BMP signaling. Blastema cells responded to all of these signals by increasing the rate of proliferation and the level of expression of the blastema marker gene, Prrx-1. The organotypic slice culture model provides the opportunity to identify and characterize the spatial and temporal co-regulation of pathways in order to induce and enhance a regenerative response.

  2. Transient developmental expression of IgY and secretory component like protein in the gut of the axolotl (Ambystoma mexicanum).

    Fellah, J S; Iscaki, S; Vaerman, J P; Charlemagne, J


    We previously reported that a primitive vertebrate, the Mexican axolotl (Amphibian, Urodela) synthesizes two classes of immunoglobulins. IgM are present in serum early in the development, and represent the bulk of specific antibody synthesis after an antigenic challenge. IgY occur in the serum later during the development, and are relatively insensitive to immunization. We demonstrate in the present work, using immunofluorescence with specific Mabs, that IgY are expressed in the gut epithelium, as secretory molecules. Secretory IgY are well expressed in the stomach and intestinal mucosae of young animals from 1 month after hatching to the seventh month. Thereafter, IgY progressively disappear from the gut and become readily detectable in the serum of 9-month-old preadult immunologically mature animals. Axolotl IgY are closely associated in the gut to secretory component-like (SC) molecules that are well-recognized by antisera to the SC of different mammalian species. This is the first description, in a primitive tetrapode, of an immunoglobulin class that could be the physiological counterpart of mammalian IgA. PMID:1627950

  3. Microhabitat Types Promote the Genetic Structure of a Micro-Endemic and Critically Endangered Mole Salamander (Ambystoma leorae) of Central Mexico

    Armando Sunny; Octavio Monroy-Vilchis; Carlos Reyna-Valencia; Martha M. Zarco-González


    The reduced immigration and emigration rates resulting from the lack of landscape connectivity of patches and the hospitality of the intervening matrix could favor the loss of alleles through genetic drift and an increased chance of inbreeding. In order for isolated populations to maintain sufficient levels of genetic diversity and adapt to environmental changes, one important conservation goal must be to preserve or reestablish connectivity among patches in a fragmented landscape. We studied...

  4. Cloning and modeling of CD8 beta in the amphibian ambystoma Mexicanum. Evolutionary conserved structures for interactions with major histocompatibility complex (MHC) class I molecules.

    Fellah, Julien S; Tuffèry, Pierre; Etchebest, Catherine; Guillet, Françoise; Bleux, Christian; Charlemagne, Jacques


    Mammalian and avian T-cells exhibit a large number of well characterized surface molecules associated with their maturation degree. Very little is known in comparison with T-cell differentiation in ectothermic vertebrates. This is mainly due to the lack of probes to identify T-cell subsets. We cloned and sequenced the first ectothermic CD8 beta DNA complementary to RNA from an amphibian species, the Mexican axolotl. The CD8 beta chain was 30-36% identical with its avian and mammalian homologues. The extracellular V-like domain contained the two typically conserved cysteines and was followed by a J-like sequence containing the canonical Phe-Gly-X-Gly stretch. The connecting peptide was much longer than in other species and contained potential O-glycosylation sites. The axolotl CD8 beta and major histocompatibility complex class I molecules were modeled using human HLA-A2/CD8 alphaalpha complex as template. The backbone conformation of axolotl CD8 beta matched well with the CD8 alpha-2 subunit of the human complex but significant structural differences were located in the CDR1, CDR2 and DE loops. Both axolotl and human class I showed large negative surface potential. The interacting area of the human CD8 alpha chain and of the corresponding region of axolotl CD8 beta had positive electrostatic potential compatible with complexation with the corresponding class I molecules. The presence of a CD8 beta homologue in an amphibian species implies that it was already present in the Devonian ancestor of amphibians and mammals, i.e. more than 400 million years ago. PMID:12034498

  5. Phylogeny of immunoglobulin heavy chain isotypes: structure of the constant region of Ambystoma mexicanum upsilon chain deduced from cDNA sequence.

    Fellah, J S; Kerfourn, F; Wiles, M V; Schwager, J; Charlemagne, J


    An RNA polymerase chain reaction strategy was used to amplify and clone a cDNA segment encoding for the complete constant part of the axolotl IgY heavy (C upsilon) chain. C upsilon is 433 amino acids long and organized into four domains (C upsilon 1-C upsilon 4); each has the typical internal disulfide bond and invariant tryptophane residues. Axolotl C upsilon is most closely related to Xenopus C upsilon (40% identical amino acid residues) and C upsilon 1 shares 46.4% amino acid residues among these species. The presence of additional cysteines in C upsilon 1 and C upsilon 2 domains is consistent with an additional intradomain S-S bond similar to that suggested for Xenopus C upsilon and C chi, and for the avian C upsilon and the human C epsilon. C upsilon 4 ends with the Gly-Lys dipeptide characteristic of secreted mammalian C gamma 3, human C epsilon 4, and avian and anuran C upsilon 4, and contains the consensus [G/GT(AA)] nucleotide splice signal sequence for joining C upsilon 4 to the transmembrane region. These results are consistent with the hypothesis of an ancestral structural relationship between amphibian, avian upsilon chains, and mammalian epsilon chains. However, these molecules have different biological properties: axolotl IgY is secretory Ig, anuran and avian IgY behave like mammalian IgG, and mammalian IgE is implicated in anaphylactic reactions. PMID:8344718

  6. Separating Population Structure from Population History: A Cladistic Analysis of the Geographical Distribution of Mitochondrial DNA Haplotypes in the Tiger Salamander, Ambystoma Tigrinum

    Templeton, A R; Routman, E.; Phillips, C A


    Nonrandom associations of alleles or haplotypes with geographical location can arise from restricted gene flow, historical events (fragmentation, range expansion, colonization), or any mixture of these factors. In this paper, we show how a nested cladistic analysis of geographical distances can be used to test the null hypothesis of no geographical association of haplotypes, test the hypothesis that significant associations are due to restricted gene flow, and identify patterns of significant...

  7. Dicty_cDB: Contig-U02845-1 [Dicty_cDB

    Full Text Available d yeast cells from Paracocci... 44 9.7 1 ( CN051673 ) v8_p6_f11_triplex_5.1ld_ V6 Ambystoma mexican...1 ( AC186951 ) Loxodonta africana clone VMRC15-414C10, WORKING D... 44 9.7 1 ( AC186328 ) Zea mays c... 44 9.7 1 ( CN051532 ) v8_p5_f17_triplex_5ld_ V6 Ambystoma cDN... 44 9.7 1 ( CN051390 ) v8_p2_e1_triplex_5ld_ V6 Ambystoma mexicanum cDNA... 44 9.7 1 ( CN051132 ) v8_p16_e9_triplex..._5.1ld_ V6 Ambystoma mexicanum c... 44 9.7 1 ( CN051110 ) v8_p16_c24_triplex_5.1l V6 Ambystoma mexican

  8. 78 FR 66058 - Habitat Conservation Plan for South Sacramento County, California


    ..., 2008 (73 FR 32729). The draft EIS/ EIR will evaluate the impacts of several alternatives related to the... Game code section 1600, and the Porter-Cologne Act, and other applicable law, on a project-by-project...) California tiger salamander, central California distinct population segment (Ambystoma californiense)...

  9. The Mexican Axolotl in Schools

    Thomas, R. M.


    Suggests and describes laboratory activities in which the Mexican axolotl (Ambystoma mexicanum Shaw) is used, including experiments in embryology and early development, growth and regeneration, neoteny and metamorphosis, genetics and coloration, anatomy and physiology, and behavior. Discusses care and maintenance of animals. (CS)

  10. Amphibian malformations and inbreeding

    Williams, Rod N.; Bos, David H; Gopurenko, David; DeWoody, J. Andrew


    Inbreeding may lead to morphological malformations in a wide variety of taxa. We used genetic markers to evaluate whether malformed urodeles were more inbred and/or had less genetic diversity than normal salamanders. We captured 687 adult and 1259 larval tiger salamanders (Ambystoma tigrinum tigrinum), assessed each individual for gross malformations, and surveyed genetic variation among malformed and normal individuals using both cytoplasmic and nuclear markers. The most common malformations...

  11. Ancient DNA Assessment of Tiger Salamander Population in Yellowstone National Park

    McMenamin, Sarah K.; Hadly, Elizabeth A.


    Recent data indicates that blotched tiger salamanders (Ambystoma tigrinum melanostictum) in northern regions of Yellowstone National Park are declining due to climate-related habitat changes. In this study, we used ancient and modern mitochondrial haplotype diversity to model the effective size of this amphibian population through recent geological time and to assess past responses to climatic changes in the region. Using subfossils collected from a cave in northern Yellowstone, we analyzed >...

  12. Dicty_cDB: Contig-U09430-1 [Dicty_cDB

    Full Text Available is chromosom... 52 9e-05 AJ004696_2( AJ004696 |pid:none) Obertrumia georgiana DNA for histo... 49 9e-05 AJ00...4697_2( AJ004697 |pid:none) Obertrumia georgiana DNA for histo... 48 2e-04 AF008301_1( AF008301 |pid:none) A...mbystoma mexicanum histone H4 (H4... 50 3e-04 AJ004695_2( AJ004695 |pid:none) Obertrumia georgiana DNA for h

  13. Hyperbaric oxygen therapy in a true regenerative environment, the regenerating limb of the axolotl

    Hansen, Kasper; Lauridsen, Henrik; Pedersen, Michael


    Introduction: Regeneration is a widespread phenomenon functioning to maintain and restore normal form and function of cells, tissues, and sometimes organs or appendages. Mammalian species typically employed in regenerative research generally process limited regenerative potential. Conversely, lower...... vertebrates such as the urodele amphibians (salamanders and newts), are excellent animal models for regenerative studies. The iconic Mexican axolotl (Ambystoma mexicanum) is capable of regenerating whole limbs, tail, jaw, and many inner organs, by dedifferentiation of cells to form a blastema (collection of...

  14. Relationships between trout stocking and amphibians in British Columbia's Southern Interior lakes

    Hirner, Joanna Lynne McGarvie


    Stocking lakes with non-native trout to encourage recreational fishing causes changes in lake ecosystems that can negatively affect biodiversity. I examined associations between rainbow trout (Oncorhynchus mykiss) and amphibians in small lakes of British Columbia’s Southern Interior by comparing abundance, growth, and probability of presence of aquatic breeding amphibians between lakes with and without trout. My evidence suggests that abundance of long-toed salamander (Ambystoma macrodactylum...

  15. The Genetics of Colonization in Two Amphibian Species After the 1980 Eruption of Mount St. Helens

    Bakkegard, Kristin Ann


    The genetics of colonization is understudied in salamanders but has large conservation implications as new habitats are formed or restored to their previous condition. The 1980 eruption of Mount St. Helens provided a natural experiment to study the genetic effects of a large infrequent environmental disturbance on two species of salamander, Taricha granulosa (Rough-skinned newt) and Ambystoma gracile (Northwestern salamander). Both these species breed in ponds, and are thought to exhibit high...

  16. Learning by embryos and the ghost of predation future

    Mathis, Alicia; Ferrari, Maud C. O.; Windel, Nathan; Messier, François; Chivers, Douglas P.


    Most research on the effects of exposure to stressful stimuli during embryonic development has focused on post-embryonic behaviour that appears to be abnormal or maladaptive. Here, we tested whether exposure to some stressful stimuli (predatory cues) can lead to post-embryonic behaviour that is adaptive. When eggs of ringed salamanders (Ambystoma annulatum) were exposed to chemical cues from predators, post-hatching larvae showed reduced activity and greater shelter-seeking behaviour; larvae ...

  17. Localization of calmodulin in epidermis and skin glands: a comparative immunohistological investigation in different vertebrate species.

    Wollina, U; Wevers, A; Mahrle, G


    The study deals with the immunolocalization of calmodulin-reactive epithelial cells in different vertebrates (Tinca tinca, Ambystoma mexicanum, Xenopus laevis, Rana ridibunda, Columba domestica, Sus scrofa domestica, Homo sapiens sapiens). The immunoperoxidase technique was performed on acetone fixed frozen sections using monoclonal (BF8) and polyclonal (ACAM) anti-calmodulin antibodies. We were able to differentiate 2 major types of staining patterns: 1. A more superficial epidermal staining in species adapted to an aqueous environment and 2. a staining along the epidermal-dermal junction in species adapted to a terrestrial environment. It seems most likely that epithelial cells immunoreactive for calmodulin are involved in skin permeability control. PMID:1718122

  18. Radioautographic investigation of retinal growth in mature amphibians

    Svistunov, S.A.; Mitashov, V.I.


    Growth of the retina was studied in mature intact amphibians, tritons, axolotls, ambystomas and clawed frogs, for six months using multiple injection of /sup 3/H-thymidine. It was established that the source of replenishment of the retina by new cells is its terminal zone in all animals investigated. This is attested to by the gradual migration of labeled cells from the growth zone into differentiated layers of the retina. The most intensely labeled cells occupy a distal position relative to other labeled cells, therefore marking the boundary between the initial part of the retina, not containing labeled nuclei, and the part being augmented. For each species studied, a level of proliferative activity is characteristic for cells of the terminal zone, which decreases in the order axolotl-clawed frog-triton -ambystoma. In the axolotl and additional growth zone is noted in the retina, in addition to the terminal, which is located in the area of the unclosed section of the embryonic fissure. Results obtained serve as a basis for the regenerative potentials of eye tissues revealed previously in these amphibian species.

  19. Germline transgenic methods for tracking cells and testing gene function during regeneration in the axolotl.

    Khattak, Shahryar; Schuez, Maritta; Richter, Tobias; Knapp, Dunja; Haigo, Saori L; Sandoval-Guzmán, Tatiana; Hradlikova, Kristyna; Duemmler, Annett; Kerney, Ryan; Tanaka, Elly M


    The salamander is the only tetrapod that regenerates complex body structures throughout life. Deciphering the underlying molecular processes of regeneration is fundamental for regenerative medicine and developmental biology, but the model organism had limited tools for molecular analysis. We describe a comprehensive set of germline transgenic strains in the laboratory-bred salamander Ambystoma mexicanum (axolotl) that open up the cellular and molecular genetic dissection of regeneration. We demonstrate tissue-dependent control of gene expression in nerve, Schwann cells, oligodendrocytes, muscle, epidermis, and cartilage. Furthermore, we demonstrate the use of tamoxifen-induced Cre/loxP-mediated recombination to indelibly mark different cell types. Finally, we inducibly overexpress the cell-cycle inhibitor p16 (INK4a) , which negatively regulates spinal cord regeneration. These tissue-specific germline axolotl lines and tightly inducible Cre drivers and LoxP reporter lines render this classical regeneration model molecularly accessible. PMID:24052945

  20. Dicty_cDB: Contig-U00933-1 [Dicty_cDB

    Full Text Available ductor muscle; &U55046_1(U55046... 51 1e-05 AJ287190_1( AJ287190 |pid:none) Radiomyces spectabilis partial ac...e) Lepeophtheirus salmonis Pacific fo... 51 1e-05 AF276076_1( AF276076 |pid:none) Ambystoma mexicanum cardiac ac...1e-05 AB179839_1( AB179839 |pid:none) Seriola quinqueradiata ACTB mRNA f... 51 1e-05 DQ666083_1( DQ666083 |pid:none) Artemia franci...83042_1( AB283042 |pid:none) Orgyia thyellina OtAct mRNA for cy... 51 1e-05 AY986487_1( AY986487 |pid:none) Rana catesbeiana cardiac...5 ) RecName: Full=Actin-104; Flags: Fragment; &U60494_1(U6... 51 1e-05 EF063572_1( EF063572 |pid:none) Actinidia delicio

  1. Dicty_cDB: Contig-U13841-1 [Dicty_cDB

    Full Text Available 32366_1( AK132366 |pid:none) Mus musculus 15 days embryo head c... 82 5e-14 BC120023_1( BC120023 |pid:none) Bos taurus tran...sp. (strain... 42 0.031 AE017352_159( AE017352 |pid:none) Cryptococcus neoformans var. neo... 42 0.031 AM778...C1099... 37 1.3 AM494961_234( AM494961 |pid:none) Leishmania braziliensis chromoso... 37 1.3 CP000686_3296( ...7 AE017348_264( AE017348 |pid:none) Cryptococcus neoformans var. neo... 35 3.7 CU...35 4.9 AM494972_271( AM494972 |pid:none) Leishmania braziliensis chromoso... 35 4.9 AF277161_1( AF277161 |pid:none) Ambystoma

  2. MRI tracking of SPIO labelled stem cells in a true regenerative environment, the regenerating limb of the axolotl

    Lauridsen, Henrik; Foldager, Casper Bindzus; Hagensen, Mette;

    Introduction: Regeneration is a widespread phenomenon functioning to maintain and restore normal form and function of cells, tissues, and in some cases organs or appendages. While mammals like mice and rats are typically employed as experimental models in regenerative research, these animals are...... generally restricted by their limited regenerative potential. Conversely, excellent animal models for regenerative studies exist in lower vertebrates such as the urodele amphibians (salamanders and newts), exemplified in the iconic Mexican axolotl (Ambystoma mexicanum) capable of regenerating whole limbs...... regenerating axolotl limb model. Superparamagnetic iron oxide particles (SPIOs) sensitive to MRI were used to track cells, and cell viability and regenerative capacity was investigated. Materials and Methods: Limb regeneration was induced by amputation of one hind limb of anaesthetised axolotls. The potential...

  3. Ion transport by the amphibian primary ureter

    Møbjerg, Nadja


    Three kidney systems appear during vertebrate development - the pronephros, mesonephros and metanephros. A characteristic of vertebrate organogenesis is the development of a primary ureter in association with the pronephros. This duct forms the collecting duct system of the latter kidneys and it is......+] steps from 3 to 20 mmol/l and a hyperpolarization of Vm upon lowering [Na+] from 102 to 2 mmol/l, indicating the presence of luminal K+ and Na+ conductances. This study provides the first functional data on the vertebrate primary ureter. The data show that the primary ureter of axolotl larvae...... putative ion transport mechanisms in the primary ureter of the freshwater amphibian Ambystoma mexicanum (axolotl). Primary ureters isolated from axolotl larvae were perfused in vitro and single cells were impaled across the basal cell membrane with glass microelectrodes. In 42 cells the membrane potential...

  4. Dicty_cDB: Contig-U03370-1 [Dicty_cDB

    Full Text Available comple... 36 0.41 A84578( A84578 ) probable WD-40 repeat protein [imported] - cDNA clo... 86 2e-21 2 ( FG985873 ) CF_WO0_86D10_SP6 Copepod Whole Organism, Nor...r_6_L01.ab1 AG Ambystoma tigrinum tigrin... 113 6e-21 1 ( EL966238 ) CF_WO0_54f03_SP6 Copepod Whole Organism, Nor...MVLFVGRVRPQYQFVYCIILAFWLGNF RQPLDSETGPF*gmrhnevi*kgsvflpyfiflidhsffng Frame B: cpacrcrsgsglfgcfvisktrts...) /CSM_Contig/Conti... 28 8.2 >Contig-U03370-1 (Contig-U03370-1Q) /CSM_Contig/Contig-U03370-1Q.Seq.d Length = 484 Score = 959 bits

  5. Comparative Analysis of Cartilage Marker Gene Expression Patterns during Axolotl and Xenopus Limb Regeneration.

    Kazumasa Mitogawa

    Full Text Available Axolotls (Ambystoma mexicanum can completely regenerate lost limbs, whereas Xenopus laevis frogs cannot. During limb regeneration, a blastema is first formed at the amputation plane. It is thought that this regeneration blastema forms a limb by mechanisms similar to those of a developing embryonic limb bud. Furthermore, Xenopus laevis frogs can form a blastema after amputation; however, the blastema results in a terminal cone-shaped cartilaginous structure called a "spike." The causes of this patterning defect in Xenopus frog limb regeneration were explored. We hypothesized that differences in chondrogenesis may underlie the patterning defect. Thus, we focused on chondrogenesis. Chondrogenesis marker genes, type I and type II collagen, were compared in regenerative and nonregenerative environments. There were marked differences between axolotls and Xenopus in the expression pattern of these chondrogenesis-associated genes. The relative deficit in the chondrogenic capacity of Xenopus blastema cells may account for the absence of total limb regenerative capacity.

  6. From biomedicine to natural history research: EST resources for ambystomatid salamanders

    Bryant Susan V


    Full Text Available Abstract Background Establishing genomic resources for closely related species will provide comparative insights that are crucial for understanding diversity and variability at multiple levels of biological organization. We developed ESTs for Mexican axolotl (Ambystoma mexicanum and Eastern tiger salamander (A. tigrinum tigrinum, species with deep and diverse research histories. Results Approximately 40,000 quality cDNA sequences were isolated for these species from various tissues, including regenerating limb and tail. These sequences and an existing set of 16,030 cDNA sequences for A. mexicanum were processed to yield 35,413 and 20,599 high quality ESTs for A. mexicanum and A. t. tigrinum, respectively. Because the A. t. tigrinum ESTs were obtained primarily from a normalized library, an approximately equal number of contigs were obtained for each species, with 21,091 unique contigs identified overall. The 10,592 contigs that showed significant similarity to sequences from the human RefSeq database reflected a diverse array of molecular functions and biological processes, with many corresponding to genes expressed during spinal cord injury in rat and fin regeneration in zebrafish. To demonstrate the utility of these EST resources, we searched databases to identify probes for regeneration research, characterized intra- and interspecific nucleotide polymorphism, saturated a human – Ambystoma synteny group with marker loci, and extended PCR primer sets designed for A. mexicanum / A. t. tigrinum orthologues to a related tiger salamander species. Conclusions Our study highlights the value of developing resources in traditional model systems where the likelihood of information transfer to multiple, closely related taxa is high, thus simultaneously enabling both laboratory and natural history research.

  7. Microarray analysis of a salamander hopeful monster reveals transcriptional signatures of paedomorphic brain development

    Putta Srikrishna


    Full Text Available Abstract Background The Mexican axolotl (Ambystoma mexicanum is considered a hopeful monster because it exhibits an adaptive and derived mode of development - paedomorphosis - that has evolved rapidly and independently among tiger salamanders. Unlike related tiger salamanders that undergo metamorphosis, axolotls retain larval morphological traits into adulthood and thus present an adult body plan that differs dramatically from the ancestral (metamorphic form. The basis of paedomorphic development was investigated by comparing temporal patterns of gene transcription between axolotl and tiger salamander larvae (Ambystoma tigrinum tigrinum that typically undergo a metamorphosis. Results Transcript abundances from whole brain and pituitary were estimated via microarray analysis on four different days post hatching (42, 56, 70, 84 dph and regression modeling was used to independently identify genes that were differentially expressed as a function of time in both species. Collectively, more differentially expressed genes (DEGs were identified as unique to the axolotl (n = 76 and tiger salamander (n = 292 than were identified as shared (n = 108. All but two of the shared DEGs exhibited the same temporal pattern of expression and the unique genes tended to show greater changes later in the larval period when tiger salamander larvae were undergoing anatomical metamorphosis. A second, complementary analysis that directly compared the expression of 1320 genes between the species identified 409 genes that differed as a function of species or the interaction between time and species. Of these 409 DEGs, 84% exhibited higher abundances in tiger salamander larvae at all sampling times. Conclusions Many of the unique tiger salamander transcriptional responses are probably associated with metamorphic biological processes. However, the axolotl also showed unique patterns of transcription early in development. In particular, the axolotl showed a genome

  8. Do Ecological Niche Models Accurately Identify Climatic Determinants of Species Ranges?

    Searcy, Christopher A; Shaffer, H Bradley


    Defining species' niches is central to understanding their distributions and is thus fundamental to basic ecology and climate change projections. Ecological niche models (ENMs) are a key component of making accurate projections and include descriptions of the niche in terms of both response curves and rankings of variable importance. In this study, we evaluate Maxent's ranking of environmental variables based on their importance in delimiting species' range boundaries by asking whether these same variables also govern annual recruitment based on long-term demographic studies. We found that Maxent-based assessments of variable importance in setting range boundaries in the California tiger salamander (Ambystoma californiense; CTS) correlate very well with how important those variables are in governing ongoing recruitment of CTS at the population level. This strong correlation suggests that Maxent's ranking of variable importance captures biologically realistic assessments of factors governing population persistence. However, this result holds only when Maxent models are built using best-practice procedures and variables are ranked based on permutation importance. Our study highlights the need for building high-quality niche models and provides encouraging evidence that when such models are built, they can reflect important aspects of a species' ecology. PMID:27028071

  9. Drought, deluge and declines: the impact of precipitation extremes on amphibians in a changing climate

    Walls, Susan C.; Barichivich, William J.; Brown, Mary E.


    The Class Amphibia is one of the most severely impacted taxa in an on-going global biodiversity crisis. Because amphibian reproduction is tightly associated with the presence of water, climatic changes that affect water availability pose a particularly menacing threat to both aquatic and terrestrial-breeding amphibians. We explore the impacts that one facet of climate change—that of extreme variation in precipitation—may have on amphibians. This variation is manifested principally as increases in the incidence and severity of both drought and major storm events. We stress the need to consider not only total precipitation amounts but also the pattern and timing of rainfall events. Such rainfall “pulses” are likely to become increasingly more influential on amphibians, especially in relation to seasonal reproduction. Changes in reproductive phenology can strongly influence the outcome of competitive and predatory interactions, thus potentially altering community dynamics in assemblages of co-existing species. We present a conceptual model to illustrate possible landscape and metapopulation consequences of alternative climate change scenarios for pond-breeding amphibians, using the Mole Salamander, Ambystoma talpoideum, as an example. Although amphibians have evolved a variety of life history strategies that enable them to cope with environmental uncertainty, it is unclear whether adaptations can keep pace with the escalating rate of climate change. Climate change, especially in combination with other stressors, is a daunting challenge for the persistence of amphibians and, thus, the conservation of global biodiversity.

  10. Ancient DNA assessment of tiger salamander population in Yellowstone National Park.

    Sarah K McMenamin

    Full Text Available Recent data indicates that blotched tiger salamanders (Ambystoma tigrinum melanostictum in northern regions of Yellowstone National Park are declining due to climate-related habitat changes. In this study, we used ancient and modern mitochondrial haplotype diversity to model the effective size of this amphibian population through recent geological time and to assess past responses to climatic changes in the region. Using subfossils collected from a cave in northern Yellowstone, we analyzed >700 base pairs of mitochondrial sequence from 16 samples ranging in age from 100 to 3300 years old and found that all shared an identical haplotype. Although mitochondrial diversity was extremely low within the living population, we still were able to detect geographic subdivision within the local area. Using serial coalescent modelling with Bayesian priors from both modern and ancient genetic data we simulated a range of probable population sizes and mutation rates through time. Our simulations suggest that regional mitochondrial diversity has remained relatively constant even through climatic fluctuations of recent millennia.

  11. High-elevation late Pleistocene (MIS 6-5) vertebrate faunas from the Ziegler Reservoir fossil site, Snowmass Village, Colorado

    Sertich, Joseph J. W.; Stucky, Richard K.; McDonald, H. Gregory; Newton, Cody; Fisher, Daniel C.; Scott, Eric; Demboski, John R.; Lucking, Carol; McHorse, Brianna K.; Davis, Edward B.


    The vertebrate record at the Ziegler Reservoir fossil site (ZRFS) near Snowmass Village, Colorado ranges from ~ 140 to 77 ka, spanning all of Marine Oxygen Isotope Stage (MIS) 5. The site contains at least 52 taxa of macro- and microvertebrates, including one fish, three amphibian, four reptile, ten bird, and 34 mammal taxa. The most common vertebrate is Ambystoma tigrinum (tiger salamander), which is represented by > 22,000 elements representing the entire life cycle. The mastodon, Mammut americanum, is the most common mammal, and is documented by > 1800 skeletal elements making the ZRFS one of the largest accumulations of proboscidean remains in North America. Faunas at the ZRFS can be divided into two groups, a lake-margin group dating to ~ 140-100 ka that is dominated by woodland taxa, and a lake-center group dating to ~ 87-77 ka characterized by taxa favoring more open conditions. The change in faunal assemblages occurred between MIS 5c and 5a (vertebrates were absent from MIS 5b deposits), which were times of significant environmental change at the ZRFS. Furthermore, the ZRFS provides a well-dated occurrence of the extinct Bison latifrons, which has implications for the timing of the Rancholabrean Mammal Age in the region.

  12. Cooperative inputs of Bmp and Fgf signaling induce tail regeneration in urodele amphibians.

    Makanae, Aki; Mitogawa, Kazumasa; Satoh, Akira


    Urodele amphibians have remarkable organ regeneration ability. They can regenerate not only limbs but also a tail throughout their life. It has been demonstrated that the regeneration of some organs are governed by the presence of neural tissues. For instance, limb regeneration cannot be induced without nerves. Thus, identifying the nerve factors has been the primary focus in amphibian organ regeneration research. Recently, substitute molecules for nerves in limb regeneration, Bmp and Fgfs, were identified. Cooperative inputs of Bmp and Fgfs can induce limb regeneration in the absence of nerves. In the present study, we investigated whether similar or same regeneration mechanisms control another neural tissue governed organ regeneration, i.e., tail regeneration, in Ambystoma mexicanum. Neural tissues in a tail, which is the spinal cord, could transform wound healing responses into organ regeneration responses, similar to nerves in limb regeneration. Furthermore, the identified regeneration inducer Fgf2+Fgf8+Bmp7 showed similar inductive effects. However, further analysis revealed that the blastema cells induced by Fgf2+Fgf8+Bmp7 could participate in the regeneration of several tissues, but could not organize a patterned tail. Regeneration inductive ability of Fgf2+Fgf8+Bmp7 was confirmed in another urodele, Pleurodeles waltl. These results suggest that the organ regeneration ability in urodele amphibians is controlled by a common mechanism. PMID:26703427

  13. Habitat alteration increases invasive fire ant abundance to the detriment of amphibians and reptiles

    Todd, B.D.; Rothermel, B.B.; Reed, R.N.; Luhring, T.M.; Schlatter, K.; Trenkamp, L.; Gibbons, J.W.


    Altered habitats have been suggested to facilitate red imported fire ant (Solenopsis invicta) colonization and dispersal, possibly compounding effects of habitat alteration on native wildlife. In this study, we compared colonization intensity of wood cover boards by S. invicta among four forest management treatments in South Carolina, USA: an unharvested control (>30 years old); a partially thinned stand; a clearcut with coarse woody debris retained; and a clearcut with coarse woody debris removed. Additionally, we compared dehydration rates and survival of recently metamorphosed salamanders (marbled salamanders, Ambystoma opacum, and mole salamanders, A. talpoideum) among treatments. We found that the number of wood cover boards colonized by S. invicta differed significantly among treatments, being lowest in the unharvested forest treatments and increasing with the degree of habitat alteration. Salamanders that were maintained in experimental field enclosures to study water loss were unexpectedly subjected to high levels of S. invicta predation that differed among forest treatments. All known predation by S. invicta was restricted to salamanders in clearcuts. The amount of vegetative ground cover was inversely related to the likelihood of S. invicta predation of salamanders. Our results show that S. invicta abundance increases with habitat disturbance and that this increased abundance has negative consequences for amphibians that remain in altered habitats. Our findings also suggest that the presence of invasive S. invicta may compromise the utility of cover boards and other techniques commonly used in herpetological studies in the Southeast. ?? 2007 Springer Science+Business Media B.V.

  14. Cannibalistic-morph Tiger Salamanders in unexpected ecological contexts

    McLean, Kyle I.; Stockwell, Craig A.; Mushet, David M.


    Barred tiger salamanders [Ambystoma mavortium (Baird, 1850)] exhibit two trophic morphologies; a typical and a cannibalistic morph. Cannibalistic morphs, distinguished by enlarged vomerine teeth, wide heads, slender bodies, and cannibalistic tendencies, are often found where conspecifics occur at high density. During 2012 and 2013, 162 North Dakota wetlands and lakes were sampled for salamanders. Fifty-one contained A. mavortium populations; four of these contained cannibalistic morph individuals. Two populations with cannibalistic morphs occurred at sites with high abundances of conspecifics. However, the other two populations occurred at sites with unexpectedly low conspecific but high fathead minnow [Pimephales promelas (Rafinesque, 1820)] abundances. Further, no typical morphs were observed in either of these later two populations, contrasting with earlier research suggesting cannibalistic morphs only occur at low frequencies in salamander populations. Another anomaly of all four populations was the occurrence of cannibalistic morphs in permanent water sites, suggesting their presence was due to factors other than faster growth allowing them to occupy ephemeral habitats. Therefore, our findings suggest environmental factors inducing the cannibalistic morphism may be more complex than previously thought.

  15. Status of amphibians on the Continental Divide:Surveys on a transect from Montana to Colorado, USA

    Corn, P.S.; Hossack, B.R.; Muths, E.; Patla, D.A.; Peterson, C.R.; Gallant, A.L.


    The Rocky Mountain Region of the United States Geological Survey's Amphibian Research and Monitoring Initiative is conducting monitoring of the status of amphibians on a transect that extends along the Continental Divide from Canada to Colorado and comprises four National Parks. Monitoring uses visual encounter surveys to determine site occupancy, with multiple visits to a subset of sites to estimate detection probabilities for each species. Detection probabilities were generally high (above 0.65) among species. There was a gradient in site occupancy, with most species scarce in the south and relatively common in the north. For example, Bufo boreas is close to extinction in Rocky Mountain National Park, was found at fewer than 5 % of sites in Yellowstone and Grand Teton National Parks in the middle of the transect, but occurs at approximately 10 % of sites in Glacier National Park. The salamander Ambystoma tigrinum was rare in Rocky Mountain and occurred at less than 25 % of sites at Yellowstone and Grand Teton, but A. macrodactylum occurred at more than 50 % of sites in Glacier. There are numerous differences among parks, such as latitude, climate, numbers of visitors, and human population density in the surrounding landscape. The degree to which these factors have influenced the current distribution and abundance of amphibians is unknown but should be a focus of additional research.

  16. The evolution of foraging rate across local and geographic gradients in predation risk and competition.

    Urban, Mark C; Richardson, Jonathan L


    Multiple theories predict the evolution of foraging rates in response to environmental variation in predation risk, intraspecific competition, time constraints, and temperature. We tested six hypotheses for the evolution of foraging rate in 24 spotted salamander (Ambystoma maculatum) populations from three latitudinally divergent sites using structural equation models derived from theory and applied to our system. We raised salamander larvae in a common-garden experiment and then assayed foraging rate under controlled conditions. Gape-limited predation risk from marbled salamanders solely explained foraging rate variation among populations at the southern site, which was dominated by this form of selection. However, at the middle and northern sites, populations evolved different foraging rates depending on their unique responses to local intraspecific density. The coupling of gape-limited predation risk from marbled salamanders and high intraspecific density at the middle site jointly contributed to selection for rapid foraging rate. At the northernmost site, intraspecific density alone explained 97% of the interpopulation variation in foraging rate. These results suggest that foraging rate has evolved multiple times in response to varying contributions from predation risk and intraspecific competition. Predation risk often varies along environmental gradients, and, thus, organisms might often shift evolutionary responses from minimizing predation risk to maximizing intraspecific competitive performance. PMID:26098352

  17. Non-additive response of larval ringed salamanders to intraspecific density.

    Ousterhout, Brittany H; Semlitsch, Raymond D


    Conditions experienced in early developmental stages can have long-term consequences for individual fitness. High intraspecific density during the natal period can affect juvenile and eventually adult growth rates, metabolism, immune function, survival, and fecundity. Despite the important ecological and evolutionary effects of early developmental density, the form of the relationship between natal density and resulting juvenile phenotype is poorly understood. To test competing hypotheses explaining responses to intraspecific density, we experimentally manipulated the initial larval density of ringed salamanders (Ambystoma annulatum), a pond-breeding amphibian, over 11 densities. We modeled the functional form of the relationship between natal density and juvenile traits, and compared the relative support for the various hypotheses based on their goodness of fit. These functional form models were then used to parameterize a simple simulation model of population growth. Our data support non-additive density dependence and presents an alternate hypothesis to additive density dependence, self-thinning and Allee effects in larval amphibians. We posit that ringed salamander larvae may be under selective pressure for tolerance to high density and increased efficiency in resource utilization. Additionally, we demonstrate that models of population dynamics are sensitive to assumptions of the functional form of density dependence. PMID:26683834

  18. Comparative Genomics of an Emerging Amphibian Virus.

    Epstein, Brendan; Storfer, Andrew


    Ranaviruses, a genus of the Iridoviridae, are large double-stranded DNA viruses that infect cold-blooded vertebrates worldwide. Ranaviruses have caused severe epizootics in commercial frog and fish populations, and are currently classified as notifiable pathogens in international trade. Previous work shows that a ranavirus that infects tiger salamanders throughout Western North America (Ambystoma tigrinum virus, or ATV) is in high prevalence among salamanders in the fishing bait trade. Bait ATV strains have elevated virulence and are transported long distances by humans, providing widespread opportunities for pathogen pollution. We sequenced the genomes of 15 strains of ATV collected from tiger salamanders across western North America and performed phylogenetic and population genomic analyses and tests for recombination. We find that ATV forms a monophyletic clade within the rest of the Ranaviruses and that it likely emerged within the last several thousand years, before human activities influenced its spread. We also identify several genes under strong positive selection, some of which appear to be involved in viral virulence and/or host immune evasion. In addition, we provide support for the pathogen pollution hypothesis with evidence of recombination among ATV strains, and potential bait-endemic strain recombination. PMID:26530419

  19. Sensitivity to nitrate and nitrite in pond-breeding amphibians from the Pacific Northwest, USA

    Marco, A.; Quilchano, C.; Blaustein, A.R.


    In static experiments, the authors studied the effects of nitrate and nitrate solutions on newly hatched larvae of five species of amphibians, namely Rana pretiosa, Rana aurora, Bufo boreas, Hyla regilla, and Ambystoma gracile. When nitrate or nitrite ions were added to the water, some larvae of some species reduced feeding activity, swam less vigorously, showed disequilibrium and paralysis, suffered abnormalities and edemas, and eventually died. The observed effects increased with both concentration and time, and there were significant differences in sensitivity among species. Ambrystoma gracile displayed the highest acute effect in water with nitrate and nitrite. The three ranid species had acute effects in water with nitrite. In chronic exposures, R. pretiosa was the most sensitive species to nitrates and nitrites. All species showed 15-d LC50s lower than 2 mg N-NO{sub 2{sup {minus}}}/L. For both N ions, B. boreas was the least sensitive amphibian. All species showed a high morality at the US Environmental Protection Agency-recommended limits of nitrite for warm-water fishes and a significant larval mortality at the recommended limits of nitrite concentration for drinking water. The recommended levels of nitrate for warm-water fishes were highly toxic for R. pretiosa and A. gracile larvae.

  20. Myofibril-Inducing RNA (MIR is essential for tropomyosin expression and myofibrillogenesis in axolotl hearts

    Lemanski Sharon L


    Full Text Available Abstract The Mexican axolotl, Ambystoma mexicanum, carries the naturally-occurring recessive mutant gene 'c' that results in a failure of homozygous (c/c embryos to form hearts that beat because of an absence of organized myofibrils. Our previous studies have shown that a noncoding RNA, Myofibril-Inducing RNA (MIR, is capable of promoting myofibrillogenesis and heart beating in the mutant (c/c axolotls. The present study demonstrates that the MIR gene is essential for tropomyosin (TM expression in axolotl hearts during development. Gene expression studies show that mRNA expression of various tropomyosin isoforms in untreated mutant hearts and in normal hearts knocked down with double-stranded MIR (dsMIR are similar to untreated normal. However, at the protein level, selected tropomyosin isoforms are significantly reduced in mutant and dsMIR treated normal hearts. These results suggest that MIR is involved in controlling the translation or post-translation of various TM isoforms and subsequently of regulating cardiac contractility.

  1. Identification of Conserved and Novel MicroRNAs during Tail Regeneration in the Mexican Axolotl

    Micah D. Gearhart


    Full Text Available The Mexican axolotl salamander (Ambystoma mexicanum is one member of a select group of vertebrate animals that have retained the amazing ability to regenerate multiple body parts. In addition to being an important model system for regeneration, the axolotl has also contributed extensively to studies of basic development. While many genes known to play key roles during development have now been implicated in various forms of regeneration, much of the regulatory apparatus controlling the underlying molecular circuitry remains unknown. In recent years, microRNAs have been identified as key regulators of gene expression during development, in many diseases and also, increasingly, in regeneration. Here, we have used deep sequencing combined with qRT-PCR to undertake a comprehensive identification of microRNAs involved in regulating regeneration in the axolotl. Specifically, among the microRNAs that we have found to be expressed in axolotl tissues, we have identified 4564 microRNA families known to be widely conserved among vertebrates, as well as 59,811 reads of putative novel microRNAs. These findings support the hypothesis that microRNAs play key roles in managing the precise spatial and temporal patterns of gene expression that ensures the correct regeneration of missing tissues.

  2. The Axolotl Fibula as a Model for the Induction of Regeneration across Large Segment Defects in Long Bones of the Extremities.

    Xiaoping Chen

    Full Text Available We tested the ability of the axolotl (Ambystoma mexicanum fibula to regenerate across segment defects of different size in the absence of intervention or after implant of a unique 8-braid pig small intestine submucosa (SIS scaffold, with or without incorporated growth factor combinations or tissue protein extract. Fractures and defects of 10% and 20% of the total limb length regenerated well without any intervention, but 40% and 50% defects failed to regenerate after either simple removal of bone or implanting SIS scaffold alone. By contrast, scaffold soaked in the growth factor combination BMP-4/HGF or in protein extract of intact limb tissue promoted partial or extensive induction of cartilage and bone across 50% segment defects in 30%-33% of cases. These results show that BMP-4/HGF and intact tissue protein extract can promote the events required to induce cartilage and bone formation across a segment defect larger than critical size and that the long bones of axolotl limbs are an inexpensive model to screen soluble factors and natural and synthetic scaffolds for their efficacy in stimulating this process.

  3. Genic regions of a large salamander genome contain long introns and novel genes

    Bryant Susan V


    Full Text Available Abstract Background The basis of genome size variation remains an outstanding question because DNA sequence data are lacking for organisms with large genomes. Sixteen BAC clones from the Mexican axolotl (Ambystoma mexicanum: c-value = 32 × 109 bp were isolated and sequenced to characterize the structure of genic regions. Results Annotation of genes within BACs showed that axolotl introns are on average 10× longer than orthologous vertebrate introns and they are predicted to contain more functional elements, including miRNAs and snoRNAs. Loci were discovered within BACs for two novel EST transcripts that are differentially expressed during spinal cord regeneration and skin metamorphosis. Unexpectedly, a third novel gene was also discovered while manually annotating BACs. Analysis of human-axolotl protein-coding sequences suggests there are 2% more lineage specific genes in the axolotl genome than the human genome, but the great majority (86% of genes between axolotl and human are predicted to be 1:1 orthologs. Considering that axolotl genes are on average 5× larger than human genes, the genic component of the salamander genome is estimated to be incredibly large, approximately 2.8 gigabases! Conclusion This study shows that a large salamander genome has a correspondingly large genic component, primarily because genes have incredibly long introns. These intronic sequences may harbor novel coding and non-coding sequences that regulate biological processes that are unique to salamanders.

  4. Association of amphibians with attenuation of ultraviolet-b radiation in montane ponds

    Adams, M.J.; Schindler, D.E.; Bury, B.R.


    Ambient ultraviolet-b (UV-B) radiation (280-320 nm) has increased at north-temperate latitudes in the last two decades. UV-B can be detrimental to amphibians, and amphibians have shown declines in some areas during this same period. We documented the distribution of amphibians and salmonids in 42 remote, subalpine and alpine ponds in Olympic National Park, Washington, United States. We inferred relative exposure of amphibian habitats to UV-B by estimating the transmission of 305- and 320-nm radiation in pond water. We found breeding Ambystoma gracile, A. macrodactylum and Rana cascadae at 33%, 31%, and 45% of the study sites, respectively. Most R. cascadae bred in fishless shallow ponds with relatively low transmission of UV-B. The relationship with UV-B exposure remained marginally significant even after the presence of fish was included in the model. At 50 cm water depth, there was a 55% reduction in incident 305-nm radiation at sites where breeding populations of R. cascadae were detected compared to other sites. We did not detect associations between UV-B transmission and A. gracile or A. macrodactylum. Our field surveys do not provide evidence for decline of R. cascadae in Olympic National Park as has been documented in Northern California, but are consistent with the hypothesis that the spatial distribution of R. cascadae breeding sites is influenced by exposure to UV-B. Substrate or pond depth could also be related to the distribution of R. cascadae in Olympic National Park.

  5. Limited influence of local and landscape factors on finescale gene flow in two pond-breeding amphibians.

    Coster, Stephanie S; Babbitt, Kimberly J; Cooper, Andrew; Kovach, Adrienne I


    Dispersal and gene flow within animal populations are influenced by the composition and configuration of the landscape. In this study, we evaluated hypotheses about the impact of natural and anthropogenic factors on genetic differentiation in two amphibian species, the spotted salamander (Ambystoma maculatum) and the wood frog (Lithobates sylvaticus) in a commercial forest in central Maine. We conducted this analysis at two scales: a local level, focused on factors measured at each breeding pond, and a landscape level, focused on factors measured between ponds. We investigated the effects of a number of environmental factors in six categories including Productivity, Physical, Land Composition, Land Configuration, Isolation and Location. Embryos were sampled from 56 spotted salamander breeding ponds and 39 wood frog breeding ponds. We used a hierarchical Bayesian approach in the program GESTE at each breeding pond and a random forest algorithm in conjunction with a network analysis between the ponds. We found overall high genetic connectivity across distances up to 17 km for both species and a limited effect of natural and anthropogenic factors on gene flow. We found the null models best explained patterns of genetic differentiation at a local level and found several factors at the landscape level that weakly influenced gene flow. This research indicates multiscale investigations that incorporate local and landscape factors are valuable for understanding patterns of gene flow. Our findings suggest that dispersal rates in this system are high enough to minimize genetic structuring and that current forestry practices do not significantly impede dispersal. PMID:25580642

  6. Toxicity of road salt to Nova Scotia amphibians

    The deposition of chemical pollutants into roadside wetlands from runoff is a current environmental concern. In northern latitudes, a major pollutant in runoff water is salt (NaCl), used as de-icing agents. In this study, 26 roadside ponds were surveyed for amphibian species richness and chloride concentration. Acute toxicity tests (LC50) were performed on five locally common amphibian species using a range of environmentally significant NaCl concentrations. Field surveys indicated that spotted salamanders (Ambystoma maculatum) and wood frogs (Rana sylvatica) did not occupy high chloride ponds. American toads (Bufo americanus) showed no pond preference based on chloride concentration. Acute toxicity tests showed spotted salamanders and wood frogs were most sensitive to chloride, and American toads were the least. Spring peepers (Pseudacris crucifer) and green frogs (Rana clamitans) showed intermediate sensitivities. We concluded that chloride concentrations in ponds due to application of de-icing salts, influenced community structure by excluding salt intolerant species. - Salt toxicity is presented as a mechanism affecting the distribution of amphibians and structure of amphibian communities in roadside wetlands

  7. Characterization of a multimeric polypeptide complex on the surface of thymus-derived cells in the Mexican axolotl.

    Kerfourn, F; Guillet, F; Charlemagne, J; Tournefier, A


    We previously raised a rabbit antiserum (L12) against a 38 kD polypeptide which is expressed on the surface of thymocytes and peripheral T cells of an Urodele Amphibian, the Mexican axolotl (Ambystoma mexicanum). Here we show that L12 antibodies immunoprecipitate several labelled molecules from surface iodinated axolotl spleen cells, including the 38 kD molecule, but also two polypeptides of 43 and 22 kD which are covalently linked to other elements. Another rabbit antiserum (L10) was raised against detergent-solubilized axolotl thymocyte membranes and shown to recognize the majority of thymocytes and about half of the splenocytes in immunofluorescence. In Western blotting, L10 antibodies recognized a limited number of surface polypeptides in thymocyte and splenocyte lysates, including 43, 38, and 22 kD elements. Immune complexes formed between L10 antibodies and solubilized splenocyte membranes were used to immunize BALB/c mice intrasplenically in the aim of raising MoAbs specific for axolotl T cells. Monoclonal antibody 87.16 was shown to stain in immunofluorescence 26.7% of thymocytes and 26.8% of spleen cells. This MoAb recognized a 43 kD polypeptide that can covalently associate on the T-cell surface with several other molecules to form a multimeric complex. PMID:8211000

  8. Conserved structure of amphibian T-cell antigen receptor beta chain.

    Fellah, J S; Kerfourn, F; Guillet, F; Charlemagne, J


    All jawed vertebrates possess well-differentiated thymuses and elicit T-cell-like cell-mediated responses; however, no surface T-cell receptor (TCR) molecules or TCR genes have been identified in ectothermic vertebrate species. Here we describe cDNA clones from an amphibian species, Ambystoma mexicanum (the Mexican axolotl), that have sequences highly homologous to the avian and mammalian TCR beta chains. The cloned amphibian beta chain variable region (V beta) shares most of the structural characteristics with the more evolved vertebrate V beta and presents approximately 56% amino acid identities with the murine V beta 14 and human V beta 18 families. The two different cloned axolotl beta chain joining regions (J beta) were found to have conserved all the invariant mammalian J beta residues, and in addition, the presence of a conserved glycine at the V beta-J beta junction suggests the existence of diversity elements. The extracellular domains of the two axolotl beta chain constant region isotypes C beta 1 and C beta 2 show an impressively high degree of identity, thus suggesting that a very efficient mechanism of gene correction has been in operation to preserve this structure at least from the early tetrapod evolution. The transmembrane axolotl C beta domains have been less well conserved when compared to the mammalian C beta but they do maintain the lysine residue that is thought to be involved in the charged interaction between the TCR alpha beta heterodimer and the CD3 complex. PMID:8341702




    Full Text Available Abstract.—Amphibians are important components of ecosystem function and processes; however, many populations havedeclined due to habitat loss, fragmentation and degradation. We studied the effect of wetlands ecosystem restoration onamphibian population recovery at Kankakee Sands in northwest Indiana, USA. We also tested predictions about colonizationin relation to proximity to existing nature preserves and species characteristics. Prior to restoration activities (1998, theamphibian community at Kankakee Sands consisted of fourteen populations of seven species at seven breeding sites. By 2001,this community increased to 60 populations at 26 sites; however, species richness had not increased. By 2002 the communityincreased to 143 populations of eight species at 38 sites, and by 2003 there were 172 populations of ten species at 44 sites.Abundance index values increased 15-fold from 1998-2003. These increases best fit the exponential growth model. Althoughsurvival through metamorphosis was substantial during wetter than average years (2002 and 2003, during other yearsrestored wetlands dried before larvae of most species transformed. Amphibian colonization was greatest near a naturepreserve with the greatest amphibian diversity. The earliest colonists included fossorial species and those species whose habitatincludes wet and mesic sand prairie. However, the fossorial Tiger Salamander (Ambystoma tigrinum was the last species tocolonize Kankakee Sands.

  10. Molecular cloning, characterization and tissue profiles of a HSP70 cDNA in Siberian sturgeon Acipenser baerii%西伯利亚鲟热休克蛋白HSP70 cDNA的克隆、序列分析和组织分布

    田照辉; 徐绍刚; 王巍; 胡红霞; 董颖; 宋超


    A full length cDNA encoding heat shock protein 70 (HSP70) was cloned from liver RNA in Siberian sturgeon (Acipenser baerii) ,and was found to contain 2 343 bp, including 5'untranslated region of 140 bp, 3'un-translated region of 256 bp and an open reading frame of 1 947 bp. The deduced 648 amino acids sequence with estimated molecular of 71 000 and theoretical isoelectric point of 5. 21 had high similarity with other vertebrate HSP70 genes in NCBI. Sequence analysis indicated that the HSP70 cDNA was consisted of three signature sequences IDL-GTTYS, IFDLGGGTFDVSrL and IVLVGGSTRIPKIQK belonging to HSP70 family. Phylogenetic tree revealed that Alligator mississippiensis, Xenopus laevis ,Ambystoma mexicanum and Sebrian Sturgeon showed a close genetic relationship. Using fluorescent real-time quantitative PCR.the HSP70 was expressed in the liver, gill, spleen, heart, intestine, muscle, gonad and brain of Siberian sturgeon, with the peak in the spleen and the minimum in the liver (P<0.05) , indicating that the HSP70 cDNA could be used in understanding of anti-stress mechanism in Siberian sturgeon and in genetic breeding improvement.%采用普通PCR和RACE技术克隆了西伯利亚鲟Acipenser baerii热休克蛋白HSP70 cDNA的全序列,该序列全长为2343 bp,其中5'非编码区为140 bp,3'非编码区为256 bp,可读编码框(ORF)为1947bp,编码为648个氨基酸.该氨基酸序列中含有HSP70家族的3个特征序列——IDLGTTYS、IFDLGGGTFDVSIL和ⅣLVGGSTRIPKIQK,细胞质特征性保守序列为EEVD,C端重复序列为GGMP.该cDNA序列与其它生物的HSP70 cDNA序列一样具有很高的相似性.系统发育树显示,西伯利亚鲟与非洲爪蛙蟾Xenopus laevis、密西西比短吻鳄Alligator mississippiensis、美西螈Ambystoma mexicanum的亲缘关系较近.实时定量分析结果表明,水温为17.5℃时,西伯利亚鲟肝脏、鳃、脾脏、心脏、肌肉、中肠、性腺、脑8种组织中均有HSP70表达,其中HSP70在脾脏中的表达

  11. Analysis of signal processing in vestibular circuits with a novel light-emitting diodes-based fluorescence microscope.

    Direnberger, Stephan; Banchi, Roberto; Brosel, Sonja; Seebacher, Christian; Laimgruber, Stefan; Uhl, Rainer; Felmy, Felix; Straka, Hans; Kunz, Lars


    Optical visualization of neural network activity is limited by imaging system-dependent technical tradeoffs. To overcome these constraints, we have developed a powerful low-cost and flexible imaging system with high spectral variability and unique spatio-temporal precision for simultaneous optical recording and manipulation of neural activity of large cell groups. The system comprises eight high-power light-emitting diodes, a camera with a large metal-oxide-semiconductor sensor and a high numerical aperture water-dipping objective. It allows fast and precise control of excitation and simultaneous low noise imaging at high resolution. Adjustable apertures generated two independent areas of variable size and position for simultaneous optical activation and image capture. The experimental applicability of this system was explored in semi-isolated preparations of larval axolotl (Ambystoma mexicanum) with intact inner ear organs and central nervous circuits. Cyclic galvanic stimulation of semicircular canals together with glutamate- and γ-aminobutyric acid (GABA)-uncaging caused a corresponding modulation of Ca(2+) transients in central vestibular neurons. These experiments revealed specific cellular properties as well as synaptic interactions between excitatory and inhibitory inputs, responsible for spatio-temporal-specific sensory signal processing. Location-specific GABA-uncaging revealed a potent inhibitory shunt of vestibular nerve afferent input in the predominating population of tonic vestibular neurons, indicating a considerable impact of local and commissural inhibitory circuits on the processing of head/body motion-related signals. The discovery of these previously unknown properties of vestibular computations demonstrates the merits of our novel microscope system for experimental applications in the field of neurobiology. PMID:25847143

  12. Genomic sequence of a ranavirus (family Iridoviridae) associated with salamander mortalities in North America

    Disease is among the suspected causes of amphibian population declines, and an iridovirus and a chytrid fungus are the primary pathogens associated with amphibian mortalities. Ambystoma tigrinum virus (ATV) and a closely related strain, Regina ranavirus (RRV), are implicated in salamander die-offs in Arizona and Canada, respectively. We report the complete sequence of the ATV genome and partial sequence of the RRV genome. Sequence analysis of the ATV/RRV genomes showed marked similarity to other ranaviruses, including tiger frog virus (TFV) and frog virus 3 (FV3), the type virus of the genus Ranavirus (family Iridoviridae), as well as more distant relationships to lymphocystis disease virus, Chilo iridescent virus, and infectious spleen and kidney necrosis virus. Putative open reading frames (ORFs) in the ATV sequence identified 24 genes that appear to control virus replication and block antiviral responses. In addition, >50 other putative genes, homologous to ORFs in other iridoviral genomes but of unknown function, were also identified. Sequence comparison performed by dot plot analysis between ATV and itself revealed a conserved 14-bp palindromic repeat within most intragenic regions. Dot plot analysis of ATV vs RRV sequences identified several polymorphisms between the two isolates. Finally, a comparison of ATV and TFV genomic sequences identified genomic rearrangements consistent with the high recombination frequency of iridoviruses. Given the adverse effects that ranavirus infections have on amphibian and fish populations, ATV/RRV sequence information will allow the design of better diagnostic probes for identifying ranavirus infections and extend our understanding of molecular events in ranavirus-infected cells

  13. Responses of pond-breeding amphibians to wildfire: short-term patterns in occupancy and colonization.

    Hossack, Blake R; Corn, Paul Stephen


    Wildland fires are expected to become more frequent and severe in many ecosystems, potentially posing a threat to many sensitive species. We evaluated the effects of a large, stand-replacement wildfire on three species of pond-breeding amphibians by estimating changes in occupancy of breeding sites during the three years before and after the fire burned 42 of 83 previously surveyed wetlands. Annual occupancy and colonization for each species was estimated using recently developed models that incorporate detection probabilities to provide unbiased parameter estimates. We did not find negative effects of the fire on the occupancy or colonization rates of the long-toed salamander (Ambystoma macrodactylum). Instead, its occupancy was higher across the study area after the fire, possibly in response to a large snowpack that may have facilitated colonization of unoccupied wetlands. Naive data (uncorrected for detection probability) for the Columbia spotted frog (Rana luteiventris) initially led to the conclusion of increased occupancy and colonization in wetlands that burned. After accounting for temporal and spatial variation in detection probabilities, however, it was evident that these parameters were relatively stable in both areas before and after the fire. We found a similar discrepancy between naive and estimated occupancy of A. macrodactylum that resulted from different detection probabilities in burned and control wetlands. The boreal toad (Bufo boreas) was not found breeding in the area prior to the fire but colonized several wetlands the year after they burned. Occupancy by B. boreas then declined during years 2 and 3 following the fire. Our study suggests that the amphibian populations we studied are resistant to wildfire and that B. boreas may experience short-term benefits from wildfire. Our data also illustrate how naive presence-non-detection data can provide misleading results. PMID:17708217

  14. Solution structure and phylogenetics of Prod1, a member of the three-finger protein superfamily implicated in salamander limb regeneration.

    Acely Garza-Garcia

    Full Text Available BACKGROUND: Following the amputation of a limb, newts and salamanders have the capability to regenerate the lost tissues via a complex process that takes place at the site of injury. Initially these cells undergo dedifferentiation to a state competent to regenerate the missing limb structures. Crucially, dedifferentiated cells have memory of their level of origin along the proximodistal (PD axis of the limb, a property known as positional identity. Notophthalmus viridescens Prod1 is a cell-surface molecule of the three-finger protein (TFP superfamily involved in the specification of newt limb PD identity. The TFP superfamily is a highly diverse group of metazoan proteins that includes snake venom toxins, mammalian transmembrane receptors and miscellaneous signaling molecules. METHODOLOGY/PRINCIPAL FINDINGS: With the aim of identifying potential orthologs of Prod1, we have solved its 3D structure and compared it to other known TFPs using phylogenetic techniques. The analysis shows that TFP 3D structures group in different categories according to function. Prod1 clusters with other cell surface protein TFP domains including the complement regulator CD59 and the C-terminal domain of urokinase-type plasminogen activator. To infer orthology, a structure-based multiple sequence alignment of representative TFP family members was built and analyzed by phylogenetic methods. Prod1 has been proposed to be the salamander CD59 but our analysis fails to support this association. Prod1 is not a good match for any of the TFP families present in mammals and this result was further supported by the identification of the putative orthologs of both CD59 and N. viridescens Prod1 in sequence data for the salamander Ambystoma tigrinum. CONCLUSIONS/SIGNIFICANCE: The available data suggest that Prod1, and thereby its role in encoding PD identity, is restricted to salamanders. The lack of comparable limb-regenerative capability in other adult vertebrates could be

  15. Functional characterization of the vertebrate primary ureter: Structure and ion transport mechanisms of the pronephric duct in axolotl larvae (Amphibia

    Prehn Lea R


    Full Text Available Abstract Background Three kidney systems appear during vertebrate development: the pronephroi, mesonephroi and metanephroi. The pronephric duct is the first or primary ureter of these kidney systems. Its role as a key player in the induction of nephrogenic mesenchyme is well established. Here we investigate whether the duct is involved in urine modification using larvae of the freshwater amphibian Ambystoma mexicanum (axolotl as model. Results We investigated structural as well as physiological properties of the pronephric duct. The key elements of our methodology were: using histology, light and transmission electron microscopy as well as confocal laser scanning microscopy on fixed tissue and applying the microperfusion technique on isolated pronephric ducts in combination with single cell microelectrode impalements. Our data show that the fully differentiated pronephric duct is composed of a single layered epithelium consisting of one cell type comparable to the principal cell of the renal collecting duct system. The cells are characterized by a prominent basolateral labyrinth and a relatively smooth apical surface with one central cilium. Cellular impalements demonstrate the presence of apical Na+ and K+ conductances, as well as a large K+ conductance in the basolateral cell membrane. Immunolabeling experiments indicate heavy expression of Na+/K+-ATPase in the basolateral labyrinth. Conclusions We propose that the pronephric duct is important for the subsequent modification of urine produced by the pronephros. Our results indicate that it reabsorbs sodium and secretes potassium via channels present in the apical cell membrane with the driving force for ion movement provided by the Na+/K+ pump. This is to our knowledge the first characterization of the pronephric duct, the precursor of the collecting duct system, which provides a model of cell structure and basic mechanisms for ion transport. Such information may be important in understanding

  16. Aquatics task force on environmental assessment of the Atikokn Power Plant: effects on aquatic organisms

    Hendrey, G.R.


    Possible impacts of an 800-MW coal-fired power plant to be built near Atikokan, Ontario were evaluated. It is feared that the emissions of SO/sub 2/ will lead to the deposition of sulfuric acid and result in the acidification of freshwaters in nearby parks and wilderness areas. The most obvious biological effects of acidification are damages to populations of fish. Less conspicuous but no less severe damages also occur to other organisms. It appears that all trophic levels are affected: species numbers are reduced, biomasses are altered, and primary production and decomposition are impaired. Field experiments and laboratory experiments indicate that microbial activity is reduced and that the recycling of materials is greatly impeded at low pH. This may interfere with nutrient supplies to plants and decrease the microbial biomass available to higher trophic levels. Phytoplankton densities decrease in acidified lakes and there is a reduction in some species of macrophytes. On the other hand, Sphagnum and benthic filamentous algae greatly increase in acidified conditions. The total primary productivity of lakes and streams may actually increase because of such dense growths on the bottom. Zooplankton and benthic invertebrate communities become less complex as acidity increases. This may in part be due to reduced food supplies, but direct inhibition by H/sub 2/SO/sub 4/ has also been demonstrated. This removal of fish food organisms may exacerbate damage to fisheries, especially in the pH range of 5 to 6. When a lake loses all fish because of low pH, a few species of invertebrates may become very abundant. The salamanders Ambystoma jeffersonium and A. maculatum, sensitive to acidity below pH 7.0 and 5.0 respectively, are being eliminated from small ponds or temporary pools in the region around Ithaca, NY because of the impact of acid precipitation. Species of frogs in some lakes are also being eliminated because of acidification. (ERB)

  17. Precise control of miR-125b levels is required to create a regeneration-permissive environment after spinal cord injury: a cross-species comparison between salamander and rat

    Juan Felipe Diaz Quiroz


    Full Text Available Most spinal cord injuries lead to permanent paralysis in mammals. By contrast, the remarkable regenerative abilities of salamanders enable full functional recovery even from complete spinal cord transections. The molecular differences underlying this evolutionary divergence between mammals and amphibians are poorly understood. We focused on upstream regulators of gene expression as primary entry points into this question. We identified a group of microRNAs (miRNAs that are conserved between the Mexican axolotl salamander (Ambystoma mexicanum and mammals but show marked cross-species differences in regulation patterns following spinal cord injury. We found that precise post-injury levels of one of these miRNAs (miR-125b is essential for functional recovery, and guides correct regeneration of axons through the lesion site in a process involving the direct downstream target Sema4D in axolotls. Translating these results to a mammalian model, we increased miR-125b levels in the rat through mimic treatments following spinal cord transection. These treatments downregulated Sema4D and other glial-scar-related genes, and enhanced the animal’s functional recovery. Our study identifies a key regulatory molecule conserved between salamander and mammal, and shows that the expression of miR-125b and Sema4D must be carefully controlled in the right cells at the correct level to promote regeneration. We also show that these molecular components of the salamander’s regeneration-permissive environment can be experimentally harnessed to improve treatment outcomes for mammalian spinal cord injuries.

  18. Microarray and cDNA sequence analysis of transcription during nerve-dependent limb regeneration

    Bryant Susan V


    Full Text Available Abstract Background Microarray analysis and 454 cDNA sequencing were used to investigate a centuries-old problem in regenerative biology: the basis of nerve-dependent limb regeneration in salamanders. Innervated (NR and denervated (DL forelimbs of Mexican axolotls were amputated and transcripts were sampled after 0, 5, and 14 days of regeneration. Results Considerable similarity was observed between NR and DL transcriptional programs at 5 and 14 days post amputation (dpa. Genes with extracellular functions that are critical to wound healing were upregulated while muscle-specific genes were downregulated. Thus, many processes that are regulated during early limb regeneration do not depend upon nerve-derived factors. The majority of the transcriptional differences between NR and DL limbs were correlated with blastema formation; cell numbers increased in NR limbs after 5 dpa and this yielded distinct transcriptional signatures of cell proliferation in NR limbs at 14 dpa. These transcriptional signatures were not observed in DL limbs. Instead, gene expression changes within DL limbs suggest more diverse and protracted wound-healing responses. 454 cDNA sequencing complemented the microarray analysis by providing deeper sampling of transcriptional programs and associated biological processes. Assembly of new 454 cDNA sequences with existing expressed sequence tag (EST contigs from the Ambystoma EST database more than doubled (3935 to 9411 the number of non-redundant human-A. mexicanum orthologous sequences. Conclusion Many new candidate gene sequences were discovered for the first time and these will greatly enable future studies of wound healing, epigenetics, genome stability, and nerve-dependent blastema formation and outgrowth using the axolotl model.

  19. Microarray analysis of microRNA expression during axolotl limb regeneration.

    Edna C Holman

    Full Text Available Among vertebrates, salamanders stand out for their remarkable capacity to quickly regrow a myriad of tissues and organs after injury or amputation. The limb regeneration process in axolotls (Ambystoma mexicanum has been well studied for decades at the cell-tissue level. While several developmental genes are known to be reactivated during this epimorphic process, less is known about the role of microRNAs in urodele amphibian limb regeneration. Given the compelling evidence that many microRNAs tightly regulate cell fate and morphogenetic processes through development and adulthood by modulating the expression (or re-expression of developmental genes, we investigated the possibility that microRNA levels change during limb regeneration. Using two different microarray platforms to compare the axolotl microRNA expression between mid-bud limb regenerating blastemas and non-regenerating stump tissues, we found that miR-21 was overexpressed in mid-bud blastemas compared to stump tissue. Mature A. mexicanum ("Amex" miR-21 was detected in axolotl RNA by Northern blot and differential expression of Amex-miR-21 in blastema versus stump was confirmed by quantitative RT-PCR. We identified the Amex Jagged1 as a putative target gene for miR-21 during salamander limb regeneration. We cloned the full length 3'UTR of Amex-Jag1, and our in vitro assays demonstrated that its single miR-21 target recognition site is functional and essential for the response of the Jagged1 gene to miR-21 levels. Our findings pave the road for advanced in vivo functional assays aimed to clarify how microRNAs such as miR-21, often linked to pathogenic cell growth, might be modulating the redeployment of developmental genes such as Jagged1 during regenerative processes.

  20. Characterizing the width of amphibian movements during postbreeding migration.

    Coster, Stephanie S; Veysey Powell, Jessica S; Babbitt, Kimberly J


    Habitat linkages can help maintain connectivity of animal populations in developed landscapes. However, the lack of empirical data on the width of lateral movements (i.e., the zigzagging of individuals as they move from one point to point another) makes determining the width of such linkages challenging. We used radiotracking data from wood frogs (Lithobates sylvaticus) and spotted salamanders (Ambystoma maculatum) in a managed forest in Maine (U.S.A.) to characterize movement patterns of populations and thus inform planning for the width of wildlife corridors. For each individual, we calculated the polar coordinates of all locations, estimated the vector sum of the polar coordinates, and measured the distance from each location to the vector sum. By fitting a Gaussian distribution over a histogram of these distances, we created a population-level probability density function and estimated the 50th and 95th percentiles to determine the width of lateral movement as individuals progressed from the pond to upland habitat. For spotted salamanders 50% of lateral movements were ≤13 m wide and 95% of movements were ≤39 m wide. For wood frogs, 50% of lateral movements were ≤17 m wide and 95% of movements were ≤ 51 m wide. For both species, those individuals that traveled the farthest from the pond also displayed the greatest lateral movement. Our results serve as a foundation for spatially explicit conservation planning for pond-breeding amphibians in areas undergoing development. Our technique can also be applied to movement data from other taxa to aid in designing habitat linkages. PMID:24423254

  1. Estimating survival and breeding probability for pond-breeding amphibians: a modified robust design

    Bailey, L.L.; Kendall, W.L.; Church, D.R.; Wilbur, H.M.


    Many studies of pond-breeding amphibians involve sampling individuals during migration to and from breeding habitats. Interpreting population processes and dynamics from these studies is difficult because (1) only a proportion of the population is observable each season, while an unknown proportion remains unobservable (e.g., non-breeding adults) and (2) not all observable animals are captured. Imperfect capture probability can be easily accommodated in capture?recapture models, but temporary transitions between observable and unobservable states, often referred to as temporary emigration, is known to cause problems in both open- and closed-population models. We develop a multistate mark?recapture (MSMR) model, using an open-robust design that permits one entry and one exit from the study area per season. Our method extends previous temporary emigration models (MSMR with an unobservable state) in two ways. First, we relax the assumption of demographic closure (no mortality) between consecutive (secondary) samples, allowing estimation of within-pond survival. Also, we add the flexibility to express survival probability of unobservable individuals (e.g., ?non-breeders?) as a function of the survival probability of observable animals while in the same, terrestrial habitat. This allows for potentially different annual survival probabilities for observable and unobservable animals. We apply our model to a relictual population of eastern tiger salamanders (Ambystoma tigrinum tigrinum). Despite small sample sizes, demographic parameters were estimated with reasonable precision. We tested several a priori biological hypotheses and found evidence for seasonal differences in pond survival. Our methods could be applied to a variety of pond-breeding species and other taxa where individuals are captured entering or exiting a common area (e.g., spawning or roosting area, hibernacula).

  2. Agricultural ponds support amphibian populations

    Knutson, M.G.; Richardson, W.B.; Reineke, D.M.; Gray, B.R.; Parmelee, J.R.; Weick, S.E.


    In some agricultural regions, natural wetlands are scarce, and constructed agricultural ponds may represent important alternative breeding habitats for amphibians. Properly managed, these agricultural ponds may effectively increase the total amount of breeding habitat and help to sustain populations. We studied small, constructed agricultural ponds in southeastern Minnesota to assess their value as amphibian breeding sites. Our study examined habitat factors associated with amphibian reproduction at two spatial scales: the pond and the landscape surrounding the pond. We found that small agricultural ponds in southeastern Minnesota provided breeding habitat for at least 10 species of amphibians. Species richness and multispecies reproductive success were more closely associated with characteristics of the pond (water quality, vegetation, and predators) compared with characteristics of the surrounding landscape, but individual species were associated with both pond and landscape variables. Ponds surrounded by row crops had similar species richness and reproductive success compared with natural wetlands and ponds surrounded by nongrazed pasture. Ponds used for watering livestock had elevated concentrations of phosphorus, higher turbidity, and a trend toward reduced amphibian reproductive success. Species richness was highest in small ponds, ponds with lower total nitrogen concentrations, tiger salamanders (Ambystoma tigrinum) present, and lacking fish. Multispecies reproductive success was best in ponds with lower total nitrogen concentrations, less emergent vegetation, and lacking fish. Habitat factors associated with higher reproductive success varied among individual species. We conclude that small, constructed farm ponds, properly managed, may help sustain amphibian populations in landscapes where natural wetland habitat is rare. We recommend management actions such as limiting livestock access to the pond to improve water quality, reducing nitrogen input, and

  3. Responses of pond-breeding amphibians to wildfire: Short-term patterns in occupancy and colonization

    Hossack, B.R.; Corn, P.S.


    Wildland fires are expected to become more frequent and severe in many ecosystems, potentially posing a threat to many sensitive species. We evaluated the effects of a large, stand-replacement wildfire on three species of pond-breeding amphibians by estimating changes in occupancy of breeding sites during the three years before and after the fire burned 42 of 83 previously surveyed wetlands. Annual occupancy and colonization for each species was estimated using recently developed models that incorporate detection probabilities to provide unbiased parameter estimates. We did not find negative effects of the fire on the occupancy or colonization rates of the long-toed salamander (Ambystoma macrodactylum). Instead, its occupancy was higher across the study area after the fire, possibly in response to a large snowpack that may have facilitated colonization of unoccupied wetlands. Naïve data (uncorrected for detection probability) for the Columbia spotted frog (Rana luteiventris) initially led to the conclusion of increased occupancy and colonization in wetlands that burned. After accounting for temporal and spatial variation in detection probabilities, however, it was evident that these parameters were relatively stable in both areas before and after the fire. We found a similar discrepancy between naïve and estimated occupancy of A. macrodactylum that resulted from different detection probabilities in burned and control wetlands. The boreal toad (Bufo boreas) was not found breeding in the area prior to the fire but colonized several wetlands the year after they burned. Occupancy by B. boreas then declined during years 2 and 3 following the fire. Our study suggests that the amphibian populations we studied are resistant to wildfire and that B. boreas may experience short-term benefits from wildfire. Our data also illustrate how naïve presence–non-detection data can provide misleading results.

  4. Level of UV-B radiation influences the effects of glyphosate-based herbicide on the spotted salamander.

    Levis, Nicholas A; Johnson, Jarrett R


    Glyphosate-based herbicides are the number one pesticide in the United States and are used commonly around the world. Understanding the affects of glyphosate-based herbicides on non-target wildlife, for example amphibians, is critical for evaluation of regulations pertaining to the use of such herbicides. Additionally, it is important to understand how variation in biotic and abiotic environmental conditions, such as UV-B light regime, could potentially affect how glyphosate-based herbicides interact with non-target species. This study used artificial pond mesocosms to identify the effects of generic glyphosate-based herbicide (GLY-4 Plus) on mortality, cellular immune response, body size, and morphological plasticity of larvae of the spotted salamander (Ambystoma maculatum) under conditions that reflect moderate (UV(M)) and low (UV(L)) UV-B light regimes. Survival within a given UV-B level was unaffected by herbicide presence or absence. However, when herbicide was present, survival varied between UV-B levels with higher survival in UV(M) conditions. Herbicide presence in the UV(M) treatments also decreased body size and reduced cellular immune response. In the UV(L) treatments, the presence of herbicide increased body size and affected tail morphology. Finally, in the absence of herbicide, body size and cellular immune response were higher in UV(M) treatments compared to UV(L) treatments. Thus, the effects of herbicide on salamander fitness were dependent on UV-B level. As anthropogenic habitat modifications continue to alter landscapes that contain amphibian breeding ponds, salamanders may increasingly find themselves in locations with reduced canopy cover and increased levels of UV light. Our findings suggest that the probability of surviving exposure to the glyphosate-based herbicide used in this study may be elevated in more open canopy ponds, but the effects on other components of fitness may be varied and unexpected. PMID:25794558

  5. The effect of community composition on persistence of prey with their predators in an assemblage of pond-breeding amphibians

    Walls, S.C.; Williams, M.C.


    We examined whether the species composition of a community influences the persistence of larval Ambystoma maculatum in assemblages composed of two larger intraguild predators (A. opacum and A. jeffersonianum) and an alternative prey species (tadpoles of Rana sylvatica). We predicted a priori that A. maculatum would have higher survival in more diverse communities containing alternative species of prey and top predators (A. opacum), the latter of which may lower the abundance of intermediate predators (A. jeffersonianum) via intraguild predation. In a factorial experiment, we manipulated the presence of larval A. opacum, A. jeffersonianum, and R. sylvatica in replicated artificial ponds containing larval A. maculatum. The presence of all three species significantly depressed biomass production in A. maculatum: biomass was highest in ponds lacking the other species and was zero in ponds initially containing all four species. Tadpoles severely reduced the growth of filamentous algae in the ponds. This, in turn, may have affected the abundance of some herbivorous prey of larval salamanders, although this possibility was not tested. The presence of congeneric predators severely restricted the presence in the water column of larval A. maculatum, which otherwise exhibited significant diel patterns of activity in the absence of predators. Together, the presence of tadpoles and a predator-mediated reduction in activity patterns may have limited foraging opportunities for A. maculatum, thus exacerbating the direct impact of predation on survival in this species. These results suggest that diverse assemblages consisting of these particular species may actually inhibit, rather than promote, inclusion of A. maculatum in some communities of pond-breeding amphibians.

  6. Low detection of ranavirus DNA in wild postmetamorphic green frogs, Rana (Lithobates) clamitans, despite previous or concurrent tadpole mortality.

    Forzán, María J; Wood, John


    Ranavirus (Iridoviridae) infection is a significant cause of mortality in amphibians. Detection of infected individuals, particularly carriers, is necessary to prevent and control outbreaks. Recently, the use of toe clips to detect ranavirus infection through PCR was proposed as an alternative to the more frequently used lethal liver sampling in green frogs (Rana [Lithobates] clamitans). We attempted reevaluate the use of toe clips, evaluate the potential use of blood onto filter paper and hepatic fine needle aspirates (FNAs) as further alternatives, and explore the adequacy of using green frogs as a target-sampling species when searching for ranavirus infection in the wild. Samples were obtained from 190 postmetamorphic (≥1-yr-old) green frogs from five ponds on Prince Edward Island (PEI), Canada. Three of the ponds had contemporary or recent tadpole mortalities due to Frog Virus 3 (FV3) ranavirus. PCR testing for ranavirus DNA was performed on 190 toe clips, 188 blood samples, 72 hepatic FNAs, and 72 liver tissue samples. Only two frogs were ranavirus-positive: liver and toe clip were positive in one, liver only was positive in the other; all blood and FNAs, including those from the two positive frogs, were negative. Results did not yield a definitive answer on the efficacy of testing each type of sample, but resemble what is found in salamanders infected with Ambystoma tigrinum (rana)virus. Findings indicate a low prevalence of FV3 in postmetamorphic green frogs on PEI (≤2.78%) and suggest that green frogs are poor reservoirs (carriers) for the virus. PMID:24502715

  7. Wide tissue distribution of axolotl class II molecules occurs independently of thyroxin.

    Völk, H; Charlemagne, J; Tournefier, A; Ferrone, S; Jost, R; Parisot, R; Kaufman, J


    Unlike most salamanders, the Mexican axolotl (Ambystoma mexicanum) fails to produce enough thyroxin to undergo anatomical metamorphosis, although a "cryptic metamorphosis" involving a change from fetal to adult hemoglobins has been described. To understand to what extent the development of the axolotl hemopoietic system is linked to anatomical metamorphosis, we examined the appearance and thyroxin dependence of class II molecules on thymus, blood, and spleen cells, using both flow cytometry and biosynthetic labeling followed by immunoprecipitation. Class II molecules are present on B cells as early as 7 weeks after hatching, the first time analyzed. At this time, most thymocytes, all T cells, and all erythrocytes lack class II molecules, but first thymocytes at 17 weeks, then T cells at 22 weeks, and finally erythrocytes at 26-27 weeks virtually all bear class II molecules. Class II molecules and adult hemoglobin appear at roughly the same time in erythrocytes. These data are most easily explained by populations of class II-negative cells being replaced by populations of class II-positive cells, and they show that the hemopoietic system matures at a variety of times unrelated to the increase of thyroxin that drives anatomical metamorphosis. We found that administration of thyroxin during axolotl ontogeny does not accelerate or otherwise affect the acquisition of class II molecules, nor does administration of drugs that inhibit thyroxin (sodium perchlorate, thiourea, methimazole, and 1-methyl imidazole) retard or abolish this acquisition, suggesting that the programs for anatomical metamorphosis and some aspects of hemopoietic development are entirely separate. PMID:9510551

  8. Structure and diversity of the heavy chain VDJ junctions in the developing Mexican axolotl.

    Golub, R; Fellah, J S; Charlemagne, J


    The immune capacity of young and adult axolotls (Ambystoma mexicanum) was evaluated by examining the combinatorial and junctional diversity of the VH chain. A large number of VDJ rearrangements isolated from 2.5-, 3.5-, 10-, and 24-month-old animals were sequenced. Six JH segments were identified with the canonical structure of all known vertebrate JHs, including the conserved Trp103-Gly104-X-Gly106 motif. Four core DH-like sequences were used by most (80%) of the VDJ junctions. These G-rich sequences had structures reminiscent of the TCRB DB sequences, and were equally used in their three reading frames. About 25% of the Igh, VDJ junctions from 3.5-month-old axolotls were out of frame, but most rearrangements were in frame at 10 and 24 months, suggesting that there is active selection of the productively rearranged Igh chains in the developing animals. There was no significant difference between the size of CDR3 in young (3.5 months) and subadult (10 months) axolotls (mean: 8.5 amino acids). However, the CDR3 loop was 1 amino acid longer in 2-year-old adult animals (mean: 9.5 residues). Several pairs of identical VDJ/CDR3 sequences were shared between 3.5-month-old individually analyzed axolotls, or between groups of axolotl of different ages. These identical rearrangements might be provided by the selection of some B-cell clones important for species survival, although the probability that different 3.5-month-old axolotl larvae would produce identical junctions seems very low, considering their limited number of B cells (less than 10(5)). The high frequency of tyrosine residues and the paucity of charged residues in the axolotl CDR3 loops may explain the polyreactivity of natural antibodies, and also clarify why it is so difficult to raise specific antibodies against soluble antigens. PMID:9271630

  9. Structure and diversity of the T-cell receptor alpha chain in the Mexican axolotl.

    Fellah, J S; Kerfourn, F; Dumay, A M; Aubet, G; Charlemagne, J


    Polymerase chain reaction was used to isolate cDNA clones encoding putative T-cell receptor (TCR) alpha chains in an amphibian, the Mexican axolotl (Ambystoma mexicanum). Five TCRalpha-V chain-encoding segments were identified, each belonging to a separate family. The best identity scores for these axolotl TCRalpha-V segments were all provided by sequences belonging to the human TCRalpha-V1 family and the mouse TCRalpha-V3 and TCRalpha-V8 families. A total of 14 different TCRA-J segments were identified from 44 TCRA-V/TCRA-J regions sequenced, suggesting that a large repertoire of TCRA-J segments is a characteristic of most vertebrates. The structure of the axolotl CDR3 alpha chain loop is in good agreement with that of mammals, including a majority of small hydrophobic residues at position 92 and of charged, hydrophilic, or polar residues at positions 93 and 94, which are highly variable and correspond to the TCRA-V/J junction. This suggests that some positions of the axolotl CDR3 alpha chain loop are positively selected during T-cell differentiation, particularly around residue 93 that could be selected for its ability to makes contacts with major histocompatibility complex-associated antigenic peptides, as in mammals. The axolotl Calpha domain had the typical structure of mammalian and avian Calpha domains, including the charged residues in the TM segment that are thought to interact with other proteins in the membrane, as well as most of the residues forming the conserved antigen receptor transmembrane motif. PMID:9002443

  10. Structure, diversity and expression of the TCRdelta chains in the Mexican axolotl.

    Fellah, Julien S; André, Sébastien; Kerfourn, Fabienne; Guerci, Aline; Durand, Charles; Aubet, Geneviève; Charlemagne, Jacques


    Mammals and birds have two major populations of T cells, based on the molecular composition and biological properties of their antigen receptors (TCR). alpha beta T cells recognize antigenic peptides linked to major histocompatibility complex (MHC) molecules, and gamma delta T cells recognize native peptide or non-peptide antigens independently of MHC. Very little is known about gamma delta T cells in ectothermic vertebrates. We have cloned and characterized the TCRdelta chains of an urodele amphibian, the Mexican axolotl (Ambystoma mexicanum). The Cdelta domain is structurally similar to its mammalian homologues and the transmembrane domain is very well conserved. Four of the six Valpha regions that can associate with Calpha (Valpha2, Valpha3, Valpha5 and Valpha6) can also associate with Cdelta, but no specific Vdelta regions were found. This suggests that the axolotl TRD locus is nested within the TRA locus, as in mammals, and that this organization has been present in all tetrapod vertebrates and in the common ancestor of Lissamphibians and mammals, for over 400 million years. Two Jdelta regions were identified, but no Ddelta segments were clearly recognized at the Vdelta-Jdelta junctions. This results in shorter and less variable CDR3 loops than in other vertebrates and the size range of the Vdelta-Jdelta junctions is similar to that of mammalian immunoglobulin light chains. Equivalent quantities of TRD mRNA were found in the lymphoid organs, and in the skin and the intestines of normal and thymectomized axolotls. The analysis of several Valpha/delta6-Cdelta and Vbeta7-Cbeta junctions showed that both the TCRdelta and the TCRbeta chains were limited in diversity in thymectomized axolotls. PMID:11981822

  11. Evolution of T cell receptor genes. Extensive diversity of V beta families in the Mexican axolotl.

    Fellah, J S; Kerfourn, F; Charlemagne, J


    We have cloned 36 different rearranged variable regions (V beta) genes encoding the beta-chain of the T cell receptor in an amphibian species, Ambystoma mexicanum (the Mexican axolotl). Eleven different V beta segments were identified, which can be classified into 9 families on the basis of a minimum of 75% nucleotide identity. All the cloned V beta segments have the canonical features of known mammalian and avian V beta, including conserved residues Cys23, Trp34, Arg69, Tyr90, and Cys92. There seems to be a greater genetic distance between the axolotl V beta families than between the different V beta families of any mammalian species examined to date: most of the axolotl V beta s have fewer than 35% identical nucleotides and the less related families (V beta 4 and V beta 8) have no more than 23.2% identity (13.5% at the amino acid level). Despite their great mutual divergence, several axolotl V beta are sequence-related to some mammalian V beta genes, like the human V beta 13 and V beta 20 segments and their murine V beta 8 and V beta 14 homologues. However, the axolotl V beta 8 and V beta 9 families are not significantly related to any other V beta sequence at the nucleotide level and show limited amino acid similarity to mammalian V alpha, V kappa III, or VH sequences. The detection of nine V beta families among 35 randomly cloned V beta segments suggests that the V beta gene repertoire in the axolotl is probably larger than presently estimated. PMID:7963525

  12. Impact of forestry practices at a landscape scale on the dynamics of amphibian populations.

    Harper, Elizabeth B; Patrick, David A; Gibbs, James P


    Forest loss is a primary cause of worldwide amphibian decline. Timber harvesting in the United States has caused dramatic changes in quality and extent of forest ecosystems, and intensive forest management still occurs. Although numerous studies have documented substantial reductions in amphibian densities related to timber harvest, subsequent extinctions are rare. To better understand the population dynamics that have allowed so many amphibian species to persist in the face of widespread forest disturbance, we developed spatially explicit metapopulation models for four forest-dependent amphibian species (Lithobates sylvaticus, Ambystoma opacum, A. talpoideum, and A. maculatum) that incorporated demographic and habitat selection data derived from experiments conducted as part of the Land Use Effects on Amphibian Populations Project (LEAP). We projected local and landscape-scale population persistence under 108 different forestry practice scenarios, varying treatment (partial cut, clear-cut with coarse woody debris [CWD] removed, and clearcut with CWD retained), cut patch size (1, 10, or 50 ha), total area cut (10, 20, or 30%), and initial amphibian population size (5, 50, or 500 adult females per local breeding population). Under these scenarios, landscape-scale extinction was highly unlikely, occurring in < 1% of model runs and for only 2 of the 4 species, because landscape-scale populations were able to persist via dispersal even despite frequent local extinctions. Yet for all species, population sizes were reduced to -50% in all clear-cut scenarios, regardless of the size of harvested patches. These findings suggest that debate over timber harvesting on pool-breeding amphibian populations in the United States should focus not on questions of landscape-scale extinction but on the ecological consequences of dramatic reductions in amphibian biomass, including changes in trophic interactions, nutrient cycling, and energy transfer. Additionally, we conclude that

  13. Interactive effects of wildfire, forest management, and isolation on amphibian and parasite abundance.

    Hossack, Blake R; Lowe, Winsor H; Honeycutt, R Ken; Parks, Sean A; Corn, Paul Stephen


    Projected increases in wildfire and other climate-driven disturbances will affect populations and communities worldwide, including host-parasite relationships. Research in temperate forests has shown that wildfire can negatively affect amphibians, but this research has occurred primarily outside of managed landscapes where interactions with human disturbances could result in additive or synergistic effects. Furthermore, parasites represent a large component of biodiversity and can affect host fitness and population dynamics, yet they are rarely included in studies of how vertebrate hosts respond to disturbance. To determine how wildfire affects amphibians and their parasites, and whether effects differ between protected and managed landscapes, we compared abundance of two amphibians and two nematodes relative to wildfire extent and severity around wetlands in neighboring protected and managed forests (Montana, USA). Population sizes of adult, male long-toed salamanders (Ambystoma macrodactylum) decreased with increased burn severity, with stronger negative effects on isolated populations and in managed forests. In contrast, breeding population sizes of Columbia spotted frogs (Rana luteiventris) increased with burn extent in both protected and managed protected forests. Path analysis showed that the effects of wildfire on the two species of nematodes were consistent with differences in their life history and transmission strategies and the responses of their hosts. Burn severity indirectly reduced abundance of soil-transmitted Cosmocercoides variabilis through reductions in salamander abundance. Burn severity also directly reduced C. variabilis abundance, possibly though changes in soil conditions. For the aquatically transmitted nematode Gyrinicola batrachiensis, the positive effect of burn extent on density of Columbia spotted frog larvae indirectly increased parasite abundance. Our results show that effects of wildfire on amphibians depend upon burn extent and

  14. Complete sequence determination of a novel reptile iridovirus isolated from soft-shelled turtle and evolutionary analysis of Iridoviridae

    Wang Xiujie


    Full Text Available Abstract Background Soft-shelled turtle iridovirus (STIV is the causative agent of severe systemic diseases in cultured soft-shelled turtles (Trionyx sinensis. To our knowledge, the only molecular information available on STIV mainly concerns the highly conserved STIV major capsid protein. The complete sequence of the STIV genome is not yet available. Therefore, determining the genome sequence of STIV and providing a detailed bioinformatic analysis of its genome content and evolution status will facilitate further understanding of the taxonomic elements of STIV and the molecular mechanisms of reptile iridovirus pathogenesis. Results We determined the complete nucleotide sequence of the STIV genome using 454 Life Science sequencing technology. The STIV genome is 105 890 bp in length with a base composition of 55.1% G+C. Computer assisted analysis revealed that the STIV genome contains 105 potential open reading frames (ORFs, which encode polypeptides ranging from 40 to 1,294 amino acids and 20 microRNA candidates. Among the putative proteins, 20 share homology with the ancestral proteins of the nuclear and cytoplasmic large DNA viruses (NCLDVs. Comparative genomic analysis showed that STIV has the highest degree of sequence conservation and a colinear arrangement of genes with frog virus 3 (FV3, followed by Tiger frog virus (TFV, Ambystoma tigrinum virus (ATV, Singapore grouper iridovirus (SGIV, Grouper iridovirus (GIV and other iridovirus isolates. Phylogenetic analysis based on conserved core genes and complete genome sequence of STIV with other virus genomes was performed. Moreover, analysis of the gene gain-and-loss events in the family Iridoviridae suggested that the genes encoded by iridoviruses have evolved for favoring adaptation to different natural host species. Conclusion This study has provided the complete genome sequence of STIV. Phylogenetic analysis suggested that STIV and FV3 are strains of the same viral species belonging to the

  15. 2010 Dry and 2009 - 2010 Wet Season Branchiopod Survey Report, Site 300

    Dexter, W


    Lawrence Livermore National Laboratory (LLNL) requested that Condor Country Consulting, Inc. (CCCI) perform wet season surveys and manage the dry season sampling for listed branchiopods in two ponded locations within the Site 300 Experimental Test Site. Site 300 is located in Alameda and San Joaquin Counties, located between the Cities of Livermore and Tracy. The two pool locations have been identified for possible amphibian enhancement activities in support of the Compensation Plan for impacts tied to the Building 850 soil clean-up project. The Building 850 project design resulted in formal consultation with the U.S. Fish and Wildlife Service (USFWS) as an amendment (File 81420-2009-F-0235) to the site-wide Biological Opinion (BO) (File 1-1-02-F-0062) in the spring of 2009 and requires mitigation for the California tiger salamander (AMCA, Ambystoma californiense) and California red-legged frog (CRLF, Rana draytonii) habitat loss. Both pools contain breeding AMCA, but do not produce metamorphs due to limited hydroperiod. The pool to the southeast (Pool BC-FS-2) is the preferred site for amphibian enhancement activities, and the wetland to northwest (Pool OA-FS-1) is the alternate location for enhancement. However, prior to enhancement, LLNL has been directed by USFWS (BO Conservation Measure 17 iii) to 'conduct USFWS protocol-level branchiopod surveys to determine whether listed brachiopod species are present within the compensation area.' CCCI conducted surveys for listed branchiopods in the 2009-2010 wet season to determine the presence of federally-listed branchiopods at the two pools (previous surveys with negative findings were performed by CCCI in 2001-2002 and 2002-2003 onsite). Surveys were conducted to partially satisfy the survey requirements of the USFWS 'Interim Survey Guidelines to Permittees for Recovery Permits under Section 10(a)(1)(A) of the Endangered Species Act for the Listed Vernal Pool Branchiopods' ('Guidelines, USFWS