In the southeastern United States, ephemeral wetlands in pine flatwoods provide important habitat for amphibians and aquatic invertebrates, but extensive deforestation has destroyed or isolated many wetlands and fire suppression has altered vegetation in others. My goals were to identify how wetland hydroperiods have changed through time and to examine the effects of long-term fire suppression on aquatic communities, including Reticulated Flatwoods Salamanders (Ambystoma bishopi) and Ornate Chorus Frogs (Pseudacris ornata). Chapter 1 used a modeling approach to relate wetland hydroperiods to current climate conditions and to hindcast historic conditions. Over the past 118 years, hydroperiods were often unfavorable for A. bishopi reproduction, and in recent years hydroperiods were shortened by persistent drought. Chapters 2 and 3 focused on identifying the effects of shifting from an open, grass dominated wetland to a wetland with high canopy cover and little herbaceous vegetation. In Chapter 2, I quantified amphibian and invertebrate communities in several wetlands. A. bishopi and P. ornata tended to occupy wetlands with lower canopy cover and higher herbaceous vegetation cover. Aquatic invertebrate abundance was generally higher in wetlands with lower shrub density and lower canopy cover. In Chapter 3, I examined how a reduction in herbaceous vegetation affected tadpoles when a predatory crayfish was present using two experiments. Crayfish were effective predators of both species across all vegetation treatments and often caused nonlethal tail injury. My results suggest that managers should focus on ensuring that wetland basins regularly burn, and wetlands with longer hydroperiods should be a management priority. / Master of Science

In contrast to mammals, some fish and amphibians have retained the ability to regenerate complex body structures or organs, such as the limb, the tail, the eye lens or even parts of the heart. One major difference in the response to injury is the appearance of a mesenchymal growth zone or blastema in these regenerative species instead of the scarring seen in mammals. This blastema is thought to largely derive from the dedifferentiation of various functional cell types, such as skeletal muscle, skin and cartilage. In the case of multinucleated skeletal muscle fibres, cell cycle re-entry into S-phase as well as fragmentation into mononucleated progenitors is observed both in vitro and in vivo. In order to identify molecules that initiate dedifferentiation of cells at the wound site in amphibians we have established a cellular assay with a cultured newt myogenic cell line. Using this assay we have found a serum activity that stimulates cell cycle re-entry in differentiated multinucleated newt myotubes. The activity is present in serum of all mammalian species tested so far and, interestingly, thrombin proteolysis amplifies the activity from both serum and plasma. We think this serum factor provides a link between wounding and regeneration and its identification will be a key step in understanding the remarkable differences in wound healing between mammals and amphibians. In the course of this PhD thesis we have characterized the serum factor as a thermo-labile, pH- and proteinase K-sensitive, high molecular weight protein that is resistant to denaturing conditions such as SDS, urea or organic solvents. Surprisingly, under denaturing conditions the activity behaves as a low molecular weight protein that displays charge heterogeneity on isoelectric focusing. Using these characteristics of the serum factor we have performed a systematic investigation of commonly used protein chromatography modes and separation techniques to develop a successful purification procedure. After four column chromatography steps -- cation exchange, hydrophobic interaction, heparin affinity and size exclusion chromatography under denaturing conditions -- we have achieved a 2,000-fold purification starting from a commercially available Crude Bovine Thrombin preparation. This represents about 40,000-fold purification over bovine serum. Silver stained gels of the most purified fractions revealed ten major protein bands. In order to finally identify the cell cycle re-entry factor, we are currently analyzing the purification by quantitative mass spectrometry by correlating the abundance of tryptic peptides with activity in sequential fractions across a chromatography run.

Cannibalism is a seemingly aberrant interaction, appearing counter to the fitness of individuals. Yet cannibalism is not overly uncommon, and naturally occurs among aquatic organisms, including larval amphibians. In temporary wetlands larval amphibians are in a race to complete metamorphosis before their aquatic habitat disappears. When intraspecific competition intensifies, eating conspecifics may represent a beneficial if not necessary strategy. The research presented within this thesis aims to characterize factors that influence cannibalism within populations of larval amphibians. Wood frog tadpoles (Lithobates sylvaticus) were used to test potential benefits of cannibalism as a diet, determine if dietary quality and nutritional stress influence cannibalism, and investigate the roles of competition and chemical cues in influencing cannibalism. Larval long-toed salamanders (Ambystoma macrodactylum), and ringed salamanders (A. annulatum) were used to investigate a functional link between trophic polymorphism and cannibalism in natural populations. Results suggest that perceived increases in competition may stimulate some individuals to become less risk averse, and more aggressive, which may in turn facilitate cannibalistic behaviour. Cannibalism itself provided only conditional benefits to larval wood frogs, rather than the optimal growth that would be expected from an ideal diet. However, this may have been the result of individual variation in response to the diet and/or conspecific cues as opposed to a nutritional deficit. In conditions where tadpoles could perceive increased competition they altered their behaviour and morphology in ways that may improve their foraging success and potentially promote cannibalism. Finally, a functional link appears to exist between head morphology and cannibalism in natural wetlands. However, the appearance of this morphology appears related to conditions that may facilitate increased population densities through rapid pond drying.

Background Molecular studies of appendage regeneration have been hindered by the lack of a stable and efficient means of transferring exogenous genes. We therefore sought an efficient integrating virus system that could be used to study limb and tail regeneration in salamanders. Results We show that replication-deficient foamy virus (FV) vectors efficiently transduce cells in two different regeneration models in cell culture and in vivo. Injection of EGFP-expressing FV but not lentivirus vector particles into regenerating limbs and tail resulted in widespread expression that persisted throughout regeneration and reamputation pointing to the utility of FV for analyzing adult phenotypes in non-mammalian models. Furthermore, tissue specific transgene expression is achieved using FV vectors during limb regeneration. Conclusions FV vectors are efficient mean of transferring genes into axolotl limb/tail and infection persists throughout regeneration and reamputation. This is a nontoxic method of delivering genes into axolotls in vivo/ in vitro and can potentially be applied to other salamander species.

No description available.

Studies were conducted on a population of tiger salamanders, Ambystoma tigrinum, at Henderson Island Wildlife Management Area in Jefferson County, TN. Tests were conducted to locate the nonbreeding habitat of the salamanders and to detect any difference in fluctuating asymmetry (FA) between larval populations in a large, permanent pond and an ephemeral wetland. Drift fences were installed with pitfall traps at selected locations around each pond to determine nonbreeding habitat use by adults. Most adult salamanders were found using a blackberry (Rubus sp.) dominated old-field, a grassy field, and a shrub-grass mix field, which were all adjacent to the ponds. No statistical difference in FA between the 2 ponds.

Declines in amphibian populations have prompted study of their ecology and distribution. The purpose of this study was to survey two sites located within different physiographic and one herpetofaunal region of Northeast Tennessee, comparing species composition and activity. The first, Henderson Wetland, is in the Appalachian Ridge and Valley physiographic region. The second, John's Bog, is in the Blue Ridge. Survey methods included random walks, aural surveys, and point source collections during a 16-month period (February 1999 to May 2000). Nine caudate (Plethodontidae) and one anuran species (Ranidae) were found in John's Bog. Seven caudate (Ambystomatidae, Plethodontidae, Salamandridae) and five anuran species (Hylidae, Ranidae) were found in Henderson Wetland. Assemblages were compared using an index of community similarity. Sites differed regarding amphibians detected. Temporal activity was not compared because of different species compositions. Instead, temporal data were compared to literature. Data will be used in future amphibian studies and site management.

A robotic microscope was designed using a microcontroller to take time-lapse digital photographs of developing salamander embryos. The microcontroller operated three stepper motors to control three-axis movement accurately, and two six mega-pixel digital cameras to capture through-focus time-lapse digital pictures of six views of Ambystoma mexicanum embryos (axolotl, a salamander). The device is designed to take images every five minutes for 80 hours of early development, from fertilization to stage 20, when the neural tube closes to form the brain and spinal column. Techniques to enhance the embryo images were investigated including image fusion to get in-focus views from a stack of images. In the early embryo surface epithelial cells differentiate to form neural tissue and external skin tissue. Observing the whole embryo surface at cellular level will give a better idea of the stress and strain each cell undergoes and what physical forces are involved in cell differentiation. / May 2007

The Coeur d’Alene salamander (Plethodon idahoensis) is a species of special concern throughout its global range, which includes southeastern BC (COSEWIC, Nov 2007), Idaho, and Montana, due to disjunct populations and sensitivity to human disturbance. Within the Interior Cedar-Hemlock forest on Mount Revelstoke, BC, Coeur d’Alene salamanders occur at their highest abundance between 600 m and 800 m. Beyond the Mount Revelstoke National Park boundaries, the low-order stream habitat of this species is subject to disturbance from forestry, mining, road building, road maintenance, and watercourse diversions such as run-of-the-river hydro projects. We conducted nocturnal salamander surveys and assessed habitat characteristics on 12 Sites (750 m² – 1000 m² stream transects) along three streams from June through September 2006. Coeur d’Alene salamanders were detected from 600 m - 1000 m on seven of the 12 study sites. Relative abundance of Coeur d’Alene salamanders ranged from 0.005 ± 0.001 per m² to 0.025 ± 0.005 per m² on six sites below 950 m. Coeur d’Alene salamanders occurred at an average of 0.001 ± 0.001 per m² at 972 m, the only site above 950 m where we detected salamanders. Our capture-mark-recapture efforts of three surveys per month in June and August yielded a very low recapture rate (3.95 %). Coeur d’Alene salamanders are challenging to enumerate due to their vertical distribution within the soil and underlying geological material. Neonate, juvenile, and adult Coeur d’Alene salamanders were observed from June to September and the highest proportion of neonates occurred in June, soon after the salamanders emerged from winter hibernation. Results of a logistic regression analysis of 1-m² plots reflected the importance of fine scale habitat characteristics (quadrat gradient, boulder, cobble, moss, grass, and shrub) in addition to site-level habitat features (water volume and elevation) that in combination describe the association of Coeur d’Alene salamanders with cool and moist conditions. Coeur d’Alene salamanders appear to select streambed habitat during warm, dry periods, which may be a behavioural response to minimize dehydration during periods of activity at the surface of the forest floor.

Thesis (Ph. D.)--Rutgers University, 2009. / "Graduate Program in Literatures in English." Includes bibliographical references (p. 235-241).