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Thesis (Ph. D.)--University of Texas at Austin, 2008. / Vita. Includes bibliographical references.

The eastern spadefoot (Scaphiopus holbrookii) is an ephemeral wetland breeding amphibian that ranges from southern Florida north and westward to southeastern Missouri and northward along the Atlantic coastal plain to Massachusetts. This species is listed as either "threatened" or "endangered" in the four states (Connecticut, Massachusetts, New York, and Rhode Island) in the northeastern United States where it is known to exist. Population declines and extirpations throughout the region over the past century have been documented and are largely believed to be the result of habitat loss and/or alteration. Very limited empirical results exist on many life history attributes of S. holbrookii anywhere in its range, including movement patterns, upland habitat selection, and breeding habitat preferences. These are critical information gaps that must be filled in order to effectively conserve and manage for this rare species in the northeastern U.S. While S. holbrookii is extremely rare throughout most of the Northeast, it is locally common in specific areas of Cape Cod National Seashore, most notably in an extensive sand-dune ecosystem known as the Province Lands located at the northern terminus of the Cape Cod peninsula. During 2005 and 2006, we conducted larval trapping surveys at 102 wetlands in the Province Lands primarily to: 1) identify breeding wetlands and 2) to assess breeding habitat use and preferences of S. holbrookii with respect to a suite of selected abiotic and biotic covariates. We captured S. holbrookii larvae at 140/652 (~21.5%) trap locations and 41/102 (~40.2%) wetlands sampled. Model results identified a number of additional habitat covariates that exhibited a statistically significant relationship with larval S. holbrookii abundance including: the percent cover of 1) cranberry (positive relationship), and 2) woody shrub (positive relationship) at a 4m radius from the trap location; 3) percent woody shrub cover at the wetland-scale (negative relationship); 4) percent canopy cover at the wetland-scale (positive relationship); 5) wetland pH (positive relationship); 6) distance to the closest paved road (positive relationship); and the kernel density cover of 7) deciduous shrubland edge (positive relationship), 8) deciduous shrubland (negative relationship), 9) pine (positive relationship), and 10) open dune (positive relationship) in the uplands surrounding the study wetlands. During 2006 and 2008 we radio-tracked adult S. holbrookii in the Province Lands using surgically implanted radio-transmitters to: 1) describe movement patterns and estimate home range sizes, and 2) assess upland habitat preferences of S. holbrookii. We tracked 19/20 individuals (11 males and 8 females) and 12/20 individuals (7 males and 5 females) for at least the first 30 and 100 days post-surgery, respectively, during 2006. During 2008 we successfully tracked 15/25 individuals (10 males and 5 females) and 5/25 individuals (3 males and 2 females) for at least the first 30 and 100 days post-surgery, respectively. In addition to results obtained on a suite of movement attributes we identified a number of upland habitat preferences for S. holbrookii. Use locations were: 1) closer to the nearest deciduous shrub edge, 2) had a greater percent cover of ground-running pitch pine branches at a 1 m scale, 3) had a greater percent cover of deciduous shrubs at a 1 m scale, and 4) had a greater percent cover of reindeer lichen at a 5 m scale. Results from this research provide much needed empirical results on these critical life history attributes related to the movement and breeding ecology of S. holbrookii. These results will aid biologists and Park management staff at Cape Cod National Seashore in more effectively employing conservation and management strategies aimed at enhancing the long-term persistence probability of this regionally rare species in the Province Lands.

The aim of the diploma thesis was to find out, which of the chosen abiotic and biotic habitat parameters different species and their hybrid groups of European fire-bellied toads prefer. The subject of the thesis was also to revise the already known locations of occurrence and to find new locations in the hybrid zone of Kaplicko and Českobudějovicko. I have monitored four new founded posts and thirteen already known posts in seven locations. I evaluated two posts as ceased, because the pool in such a location has been rebuilt for fish pond management. One location has been degraded by building waste. Another one has ceased to exist because there has been built a new family house at the post. Four posts were regularly without findings because of excessive drought. My monitoring was based on the method of complete capturing and the determination was made on the basis of the pattern of aposematic coloration on the bellies of the caught toads. I also determined the degree of hybridization (hybrid index) according to this pattern. I classified nine posts in five locations in total, of which four of them have been newly found. On the basis of obtained results I decided with the help of the Pearson correlation coefficient some parameters were not confirmative: a perimeter, an area, a depth and vegetation presence. Sun exposure, forest distance and the altitude of the posts showed to be statistically significant.

Thesis (Ph. D.)--University of Alberta, 2010. / Title from pdf file main screen (viewed on June 11, 2010). A thesis submitted to the Faculty of Graduate Studies and Research in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Environmental Biology and Ecology, Department of Biological Sciences, University of Alberta. Includes bibliographical references.

The goal of my dissertation was to describe the dynamics of a group of Houston toads located at the Griffith League Ranch (GLR), Bastrop County, Texas. My research included using statistical modeling to predict activity and abundance, mark-recapture techniques to estimate survivorship, and simulation modeling to explore the impacts of the difference in age at first reproduction and to project the future dynamics of the population at the GLR. From 2001 – 2005, 225 individual Houston toads (199 M : 26 F) were captured using two methods: breeding pond surveys and drift fences. Houston toads were neither caught equally among capture methods, nor across years. Toad activity was mostly confined within their breeding season, and activity was not continuous. A logistic regression indicated activity depended on time of year, mean precipitation, mean minimum daily temperature, and mean percent lunation as well as two-way interactions with moon-phase and other variables. Abundance depended on time of year, current precipitation, minimum temperature, and two-way interactions between time of year and the other two variables. Twenty-one of the 199 males (10.5%) and no females were recaptured among years. The probability of male survival was estimated using program MARK. Eight of 16 candidate models were supported and all but one contained precipitation as a covariate, indicating precipitation is important for Houston toad survival. Survivorship estimates varied from 0.1 to 0.41. The sex ratio was significantly male-biased. The odds of catching females in traps were 3.5 greater than capturing females in a pond, while the odds of capturing males in a trap were 0.28 compared to ponds. Results from a simulation model indicated the sex ratio is biased because of the difference in maturation times between males and females, coupled with high juvenile mortality. Results from an individual-based, spatially-explicit, stochastic simulation model, indicated a relatively low probability (~ 0.013) of B. houstonensis going extinct at the GLR within the next 10 years. Emergent properties of the model were similar to results observed in the field or reported in the literature. The model also identified that dispersal of Houston toads should be a future research priority.

The goal of my dissertation was to describe the dynamics of a group of Houston toads located at the Griffith League Ranch (GLR), Bastrop County, Texas. My research included using statistical modeling to predict activity and abundance, mark-recapture techniques to estimate survivorship, and simulation modeling to explore the impacts of the difference in age at first reproduction and to project the future dynamics of the population at the GLR. From 2001 – 2005, 225 individual Houston toads (199 M : 26 F) were captured using two methods: breeding pond surveys and drift fences. Houston toads were neither caught equally among capture methods, nor across years. Toad activity was mostly confined within their breeding season, and activity was not continuous. A logistic regression indicated activity depended on time of year, mean precipitation, mean minimum daily temperature, and mean percent lunation as well as two-way interactions with moon-phase and other variables. Abundance depended on time of year, current precipitation, minimum temperature, and two-way interactions between time of year and the other two variables. Twenty-one of the 199 males (10.5%) and no females were recaptured among years. The probability of male survival was estimated using program MARK. Eight of 16 candidate models were supported and all but one contained precipitation as a covariate, indicating precipitation is important for Houston toad survival. Survivorship estimates varied from 0.1 to 0.41. The sex ratio was significantly male-biased. The odds of catching females in traps were 3.5 greater than capturing females in a pond, while the odds of capturing males in a trap were 0.28 compared to ponds. Results from a simulation model indicated the sex ratio is biased because of the difference in maturation times between males and females, coupled with high juvenile mortality. Results from an individual-based, spatially-explicit, stochastic simulation model, indicated a relatively low probability (~ 0.013) of B. houstonensis going extinct at the GLR within the next 10 years. Emergent properties of the model were similar to results observed in the field or reported in the literature. The model also identified that dispersal of Houston toads should be a future research priority.

The behavioral and physiological interaction between Bufo bankorensis and its snake predators, Macropisthodon rudis and Dinodon rufozonatum were investigated in this study. Adult M. rudis preferred medium sized toads but neonate favored to small ones. About seized position of different toad sizes, adult snakes attacked large toads head first but neonates did randomly. On the swallow direction with toad sizes, adult snakes had no preference but neonates swallowed large toads head first, so the neonates could change the direction swallowing preys after preys had been seized. Adult and neonate snakes spent similar time in handling toads with head first ingestion costed less handling time. After the large rear-teeth had been grounded, M. rudis would spend more time in handling preys. In all trials, venom secreted from parotid glands did not seem to prolong handling time on adult snake. After forced fed 1 mg/g toad venom to D. rufozonatum, no significant change of Na+, K+, Ca2+ ion concentrations were observed but significant increasing of heart rates. The ECGs of intoxicated snakes showed typical digitalis-like cardioactive effects as sinus arrhythmia, prolonged P-R interval as varying degrees of atrioventricular block and ventricular fibrillation. The LD50 of the venom of B. bankorensis are different between from toads sampled Northern and Southern Taiwan. In comparison with dissected toad-eating snakes D. rufozonatum, M. rudis and Rhabdophis tigrinus formosanus, I didn¡¦t find any nuchal glands on nuchal-dorsal site of D. rufozonatum and M. rudis.

To date, the majority of studies into sex determination and sexual development have focused on the mammalian system due to the mouse model being an excellent tool for developmental biology as well as obvious implications to human health and development. However, the focus on the mammalian system has caused a large fraction of other vertebrate groups to be overlooked. The last decade has seen an array of studies into various non-mammalian organisms like fishes, birds, reptiles, and amphibians. These studies have unveiled a remarkably conserved molecular background utilized for sexual differentiation amongst all vertebrates, ranging from the alligator which employs temperature dependent sex determination to the mouse, which employs genotypic sex determination In this project, I implemented molecular methods traditionally used to study model organisms, to investigate an amphibian species, the cane toad (Bufo marinus). The cane toad was chosen due to its invasive status in Australia, as well as being a representative of one of the most successful and specious families of vertebrates, Bufonidae. Since, its introduction, this species has rapidly spread across the continent, adversely effecting native species throughout its introduced range. Recent studies have identified the disruption of the sex-determination pathway as a realistic goal in an otherwise, seemingly futile, effort to curtail their expansion. We decided to approach the study of cane toad sex determination and development on three levels, in order to have a broader understanding of the similarities between toads and other groups, and additionally, to better assess the feasibility of utilizing molecular means to control cane toads in non-native habitats. Firstly, I used a candidate gene approach to clone and characterize five genes which were previously known to be involved in both mammalian as well as amphibian sex determination: Sox9, Dmrt1, p450arom, Sf1, and Dax1. I chose Sox9 and Dmrt1 due to their known involvement in the male-specific pathway of mammals. Dmrt1 was also known to be male specific in other groups, including amphibians. Conversely, I also decided to investigate p450arom and Dax1, due to their affiliation with the female pathway, while Sf1 has a role in both sexes. All of these genes were expressed in the gonads of both sexes of cane toads. However, Sox9 exhibited strong transcriptional up-regulation in testes at the time of sexual differentiation, similar to mammals. Thus, using the candidate gene approach, I was successful in identifying a sex-specific marker which could be utilized in the manipulation of the sex determination pathway (e.g. female to male sex reversal). VIIThe second approach involved the study of sex determination on a cytogenetic level. Traditionally, Bufonids have been assumed to utilize a ZZ/ZW (male homogametic/female heterogametic) sex chromosome system, yet only one recent study has been successful in the identification of sex chromosomes in a toad species. Moreover, the authors identified the sex chromosomes in only one population (of 18 studied), highlighting the scarcity of discernible sex chromosomes in toads. When we began to investigate the karyotype of cane toads, we were able to identify a female-specific length polymorphism in the nucleolus organizer region (NOR) of chromosome 7, making this chromosome pair a strong candidates for the Z and W sex chromosomes. In order to verify our hypothesis, we also performed chromomycin A3 staining to reveal a differential signal between the chromosome pair, indicative of heterochromatin accumulation on the brighter NOR. This study lent strong support to the observed difference between the chromosomes, yet we required further investigation in order to identify the W and the Z chromosomes, specifically. By utilizing comparative genomic hybridization (CGH), I was able to identify a female specific region on the chromosome with the larger NOR, identifying it as the W chromosome. This study was successful in identifying the sex chromosomes in the cane toad. Moreover, it allows us to make a more accurate prediction of the possible sex determination method utilized: either a dosage dependent male determination system, or a female-specific gene in a female determination system. The third approach focused on the Bidder’s organ and its role in sexual development of cane toads. The Bidder’s organ is a gonadal structure comprised of ovary-like tissue, found in both males and females, with no known function. Since our study was focused on female to male sex-reversal of cane toads, we decided to investigate the potential of the Bidder’s organ to interrupt this process. By utilizing the candidate genes, which I had previously cloned, I was able to identify the Bidder’s organ as having an entirely distinct transcriptional pattern in comparison to the gonads of either sex. Additionally, the Bidder’s organ showed significantly higher levels of p450arom expression than the gonads, identifying it as a possible key player in the production of aromatase enzyme for oestrogen production. Moreover, gene expression patterns in the Bidder’s organ strongly correlated with the significant developmental time points in the sexual development of the toads, implying a possible function in the development process of toads. In conclusion, I have performed the first study of the molecular, cytogenetic and anatomical aspects of sexual development in a toad. I was able to verify that cane toads utilize a ZZ/ZW chromosome system and furthermore, likely utilize the mammalian male-linked gene, Sox9, in male development. I was also able to show that the Bidder’s organ is transcriptionally active at key time points, likely indicative of a functional role during development.

Thesis (M.S.)--Brigham Young University. Dept. of Integrative Biology, 2006. / Includes bibliographical references (p. 27-34).