Thesis (B.A.)-Oregon State University, 1997. / Includes bibliographical referenes (leaves 20-22). Also available via the Internet.

The analysis of diverse data sets can yield different phylogenetic estimates that challenge systematists to explain the source of discordance. The Ambystomatidae are a classic example of this phylogenetic conflict. Previous attempts to resolve the ambystomatid species tree using allozymic, morphological, and mitochondrial sequence data have yielded different estimates, making it unclear which data source best approximates ambystomatid phylogeny. We present the first multi-locus DNA sequencebased phylogenetic study of the Ambystomatidae. Because independent loci can contain discordant gene tree histories, concatenating unlinked loci into a single data matrix can lead to strongly supported and erroneous results. Therefore, we utilized a range of analyses, including coalescent-based methods of phylogenetic estimation that account for incomplete lineage sorting and concordance-based methods that estimate the proportion of sampled loci that support a particular clade. We repeated these analyses with the removal of individual loci to determine if any locus has a disproportionate effect on our phylogenetic results. Many deep and relatively shallow clades within Ambystoma were robustly resolved. Analyses that excluded loci produced overlapping posterior distributions, suggesting no disproportionate influence of any particular locus. Our estimates differ from previous hypotheses, although there was greater similarity with previous molecular estimates, relative to morphological estimates.

Unlike many amphibians, the paedomorphic axolotl (Ambystoma mexicanum) rarely undergoes external morphological changes indicative of metamorphosis. However, internally, some axolotl tissues undergo cryptic metamorphic changes. A previous study examined interspecific patterns of larval brain gene expression and found that these species exhibited unique temporal expression patterns that were hypothesized to be morph specific. This thesis tested this hypothesis by examining differences in brain gene expression between juvenile (JUV), paedomorphic (PAED), and metamorphic (MET) axolotls. I identified 828 genes that were expressed differently between JUV, PAED, and MET. Expression estimates from JUV were compared to estimates from PAED and MET brains to identify genes that changed significantly during development. Genes that showed statistically equivalent expression changes across MET and PAED brains provide a glimpse at aging and maturation in an amphibian. The genes that showed statistically different expression estimates between metamorphic and paedomorphic brains provide new functional insights into the maintenance and regulation of paedomorphosis. For genes that were not commonly regulated due to aging, paedomorphs exhibited greater transcriptional similarity to juvenile than metamorphs did to juvenile. Overall, gene expression differences between metamorphic and paedomorphic development exhibit a mosaic pattern of expression as a function of aging and metamorphosis in axolotls.

Thèse (M.Sc.) -- Université Laval, 1997. / Bibliogr.: f. 98-115. Publié aussi en version électronique.

No description available.

In the Southern Appalachians, there are growing concerns about Spotted Salamanders (Ambystoma maculatum) due to habitat destruction and the uncertain future of their forested riparian habitat. In this study, I establish baseline data of a population of Spotted Salamanders so future monitoring of this population can be performed. The study site is located in northeastern Tennessee, just east of the city of Bristol. It is a vernal pond at the Osceola Island Recreation Area one mile below the South Holston Dam on Holston View Dam road. A drift fence was established completely around the perimeter of the vernal pond to capture Spotted Salamanders entering and exiting the study pond. Individual Spotted Salamanders were identified using spot pattern recognition. An estimated population size of 2,449 Spotted Salamanders was determined to utilized the pond for breeding for the spring of 2003.

Mole salamanders (Ambystoma) and woodfrogs (Lithobates sylvaticus) are abundant in New England and depend on ephemeral wetlands for breeding. Their aquatic habitats have been well studied and are protected by several local and regional regulations. State endangered species laws also protect mabled salamanders (A. opacum), Jefferson salamanders (A. jeffersonianum), and blue-spotted salamanders (A. laterale). However, these amphbibians spend most of their adult lives in terrestrial habitats that remain poorly protected and elusive to researchers. In chapter 1, I developed a novel technique using passive integrated transponders for tracking small animals. I used this technique to track marbled salamanders walking up to 200 m from their breeding pond during post-breeding migrations. In Chapter 2, I examined the importance of multiple habitat variables for predicting the distributions of woodfrogs and spotted salamanders at 455 ponds in western Massachusetts. Based on a variable-comparison technique I developed, the best predictor for either species of amphibian was the amount of forest in the surrounding vii landscape. Both species were found more frequently in upland forests where the ponds are least protected by state and federal wetland regulations. In chapter 3, I used my data from chapter 2 and three other similar data sets to conduct an analysis of spatial scale and to parameterize a recently published resistant kernel model. The complex model parameterized by an expert panel did significantly worse than the null model. The distributions of both amphibians were best predicted by measuring the landscape at very large scales (over 1000 m). The most effective scales for conservation may be largest for organisms of intermediate dispersal capability. In chapter 4, I explored the evolution and genetics of the Jefferson/blue-spotted/unisexual salamander complex. I framed research into the fascinating unisexual reproductive system with a model that relates nuclear genome replacement, positive selection on hybrids, and biogeography of the species complex. I parameterized this model using genetic data taken from salamanders spanning Massachusetts and an individual-based breeding simulation. If paternal genomes are transmitted to offspring with the frequencies reported from laboratory experiments, then my model suggests that there must be strong selection favoring unisexuals with hybrid nuclei.

No description available.

Assessing ecological risk to wildlife exposed to anthropogenic contamination in soil has traditionally been problematic. Attempts to standardize an approach to evaluate risk for various community types in North America have been challenging, given the variation in terrestrial communities and the values in which policy makers are bound to protect. This has resulted in vague, yet flexible guidance from the U.S. Environmental Protection Agency and other interested parties (e.g., the U.S. Army Corps of Engineers, and the Tri-Service Ecological Risk Assessment Working Group). Interpretation of these and other guidance has been variable, often resulting in conflicting opinions on how best to address the question of ecological risk to receptors that are exposed to xenobiotics in a soil matrix. This work reports the results of research designed to address the question of ecological risk to terrestrial vertebrates. Objective, ecologically-relevant criteria were used in the selection and development of models in this research. Several lines of logic were considered: 1) substance sensitivity, 2) ecological sensitivity (i.e., the species importance to the system; e.g., keystone species); and, 3) probability and extent of exposure. A primary soil contaminant at many U.S. Army installations is 2,4,6-trinitrotoluene (TNT). This was a result of the mass manufacturing, storing, and assembly of weapons from the early 1900's until the 1950s. The Army has reported soil concentrations of TNT ranging from 0.12 to 38,600 ug/g (Walsh and Jenkins 1992) and 0.08 to 64,000 ug/g (Hovatter et al. 1997). The chemical-physical properties of TNT result in a relatively unique compound, not easily amenable to current modeling techniques to estimate exposure to terrestrial wildlife. Moreover, there are few data describing the effects of exposure to TNT in other than mammals, fish, and specific invertebrates. In this research, the pathways of exposure and selected potential toxic effects from TNT exposure were investigated in a terrestrial salamander: Ambystoma tigrinum (tiger salamanders). A. tigrinum was chosen since they are exclusively carnivorous, relatively long-lived, have a thin integument, and are large enough to investigate individual effects. These investigations were designed to mimic natural conditions as closely as possible, though maintain a degree of homogeneity in a laboratory environment. All studies exposed salamanders to soil and food (earthworms) in identical preparations. As such, these exposures were considered complete, eliminating assumptions made regarding daily food consumption, systemic dermal dose, etc. The first study examined the relative contribution of dermal or oral exposures to the whole-body burdens of TNT and primary metabolites. A poly-chlorinated biphenyl (PCB) mixture (Aroclor7 1260) was used with TNT to simultaneously to assist in the evaluation of each pathway, since the fate and transport of PCBs are well characterized. Tiger salamanders were exposed 28 days in situ. The dermal route of exposure contributed the most to the final burdens of TNT in salamanders, with the primary reduction products, 2-amino-4,6-dinitrotoluene and 4-amino, 2,6-dinitrotoulene reaching higher concentrations than of parent compound. Other TNT metabolites were found in insignificant quantities. The concentrations of PCBs were higher in the oral treatment, as expected. These results were corroborated in a subsequent study using Ambystoma maculatum (spotted salamanders). The second series of investigations evaluated the potential toxic effects from TNT exposure. Two treatments consisting of TNT and a control were used to evaluate these effects to A. tigrinum. The salamanders were exposed in situ for 14 days to TNT in soil and food (earthworms of which were exposed to TNT in the soil in similar preparations). Non-specific immune effects were evaluated through the characterization of splenic phagocytes in their ability to: 1) phagocytize foreign particles, and 2) digest (through oxygen radicals) phagocytized material. This was conducted using fluorescent microspheres and a fluorescent chemical probe specific to hydrogen peroxide, measured per each cell using flow cytometry. Other data collected included histological examination (e.g., liver, kidney, and other miscellaneous organs), blood differentials, weight changes over time, organ/ body weight comparisons, and an analysis of organ-specific metabolism. No significant effects were noted in salamanders exposed to these conditions. Coordinated with the preceding study included a search for biomarkers of exposure and an investigation of the metabolites of TNT in situ. Biotransformation products of TNT were found including primary (e.g., 2-amino-4,6-dinitrotoluene) and secondary (e.g., 2,4-diamino-6-nitrotoluene) in relative concentrations in skin, liver, and kidney. Biomarkers of exposure included an analysis of cytochrome p450, b5, and the glutathione antioxidant enzymes in liver, kidney, skin, lung, and serum, respectively. Traces of parent compound were found in the skin and liver only. Levels of 2,4-diamino-6-nitrotoluene were found only in the liver and kidney, suggesting that TNT is reduced primarily in or on the skin. Levels of p450 were higher in TNT exposed salamanders than controls. Glutathione and related enzyme levels are reported. This work suggests that salamanders have levels of detoxification enzymes capable of the biotransformation of anthropogenic substances in soil rivaling that of mammals. Another investigation evaluated these same immunological parameters in white-footed mice (Peromyscus leucopus). This species was chosen based on the relative importance of small mammals to the community structure in many North American ecosystems. Mice were exposed to TNT in the feed at 0.264, 0.066, 0.033, and 0.017%, where actual daily dose estimates for males were 604, 275, 109, and 65; and for females was 544, 282, 143, and 70 mg/kg/d. An investigation to evaluate the specificity of commercially-available monoclonal antibodies specific to cell surface markers for thymocytes and splenocytes in inbred mice was unsuccessful. These results suggest the recognition epitopes of monoclonal antibodies prepared against Old-World mice are not conserved into Peromyscus, a New-World species. However, high dose males and females had larger spleens consistent with the hemolytic effects previously reported for mammals exposed to TNT. Further, males exposed at all levels had reduced phagocytic activity of splenocytes, and reduced hydrogen peroxide production associated with the two highest doses relative to controls. Females showed no response relative to treatment. This research has shown the feasibility for these types of investigations, and provides toxicity information valuable for modeling estimates of ecological risk. Further, the in situ exposures have provided media concentrations that are or are not toxic for species of concern. This type of information reduces the uncertainty associated with ingestion modeling estimates, dermal exposure estimates, and other factors not traditionally considered in toxicity studies. / Ph. D.

Species that are introduced outside of their native ranges are an important threat to biodiversity. In southwestern Alberta, Canada, sport and bait fish have been introduced into most waterbodies. I examined the effects of introduced trout and minnows on the distribution, demography and behaviour of larval long-toed salamanders through a combination of field surveys, laboratory experiments and an outdoor mesocosm experiment. Results from field surveys at 30 high-elevation (>1500m) lakes confirmed previous studies showing an allopatic distribution of trout and long-toed salamanders. The same pattern was also documented at 27 low-elevation (<1500m) ponds. In a mesocosm experiment, salamander survival was significantly reduced in ponds containing trout or minnows. Surprisingly, larvae exposed to minnows were 28-65% smaller than larvae in control ponds, suggesting strong interspecific competition for zooplankton prey. In a series of laboratory studies, trout preyed directly on salamander hatchlings and larvae, whereas minnows injured hatchlings but did not consume them. In laboratory aquaria, salamander larvae spent significantly more time within a refuge when exposed to minnow cues, but showed no behavioural response to trout. This confirmed my expectation that long-toed salamanders lack specific behavioural responses to trout, but respond generally to disturbances within the water column. Thus, direct predation and a lack of specific antipredator behaviour are among the likely mechanisms responsible for the observed allopatic distribution of trout and long-toed salamanders. My data also show that gape-limited fish reduce growth and survival of salamanders, perhaps more so than trout, through mechanism such as competition and behavioural alteration. / ix, 76 leaves : ill., map ; 29 cm.