Effects of commercially available bacterial products were investigated on two common recreational pool contaminants: sunscreen and cyanuric acid (CYA). Microbial products developed by BiOWiSH Technologies, Inc. were tested for enhancing mechanical filtration and water clarification in bench-scale bioreactors, with conditions mimicking those of recreational pool water. Bacterial consortia included proprietary mixes of Bacillus, Lactobacillus and Pseudomonas, and other genera of bacteria. BiOWiSH products are either fermented on a solid substrate consisting of rice bran and soy meal, or they are mixed with a soluble diluent. Twenty-nine BiOWiSH products were tested throughout forty experiments. Experiments were carried out to determine both the efficacy of BiOWiSH products for turbidity reduction and the mechanism by which BiOWiSH removes sunscreen from solution. In trials without mechanical filtration, the only product which showed a reduction in turbidity relative to the control, albeit inconsistently, was the solid substrate version of BiOWiSH Aqua FOGTM (Thai FOG). Experiments on BiOWiSH coupled with mechanical filtration showed a 79% average reduction of turbidity in the first 24 hrs. BiOWiSH products containing solid substrate, both active and abiotic, showed an average turbidity reduction of 90% in the first 24 hrs. In the same timeframe, soluble BiOWiSH products showed a 79% average reduction in turbidity. Thus, the solid v substrate provided an additional 11% reduction in turbidity over soluble products and un-amended mechanical filtration. Through experimentation and scanning electron microscopy, it was concluded that the primary mechanism of clarification by the solid substrate is adsorption of sunscreen to the substrate surface. Further experiments were performed in anaerobic and aerobic environments to determine whether BiOWiSH products can remove cyanuric acid from solution through adsorption or biodegradation. Two measurement methods, turbidimetric and HPLC (high performance liquid chromatography) were used to independently quantify CYA. A reverse-phase HPLC method was developed which utilizes a phosphate buffer and methanol for the separation of cyanuric acid from nitrate and other chemical species. The solid BiOWiSH Aqua FOG product (prod. in Thailand) interfered with the turbidimetric analysis, showing false decreases in CYA. Using HPLC, there was no measureable biodegradation or adsorption of CYA by BiOWiSH products in these bench-scale tests. Significant systematic error in the HPLC analysis prevented conclusive findings; therefore, the ability of BiOWiSH products to reduce CYA from solution remains inconclusive.

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.

Vernal pools are essential breeding habitat for amphibians - the vertebrates most at-risk across the globe. Unfortunately, due to their small sizes and temporary nature, vernal pools are prone to indiscriminate destruction. This is the case in southern Ontario as most vernal pools have already been destroyed by human development. As such there is an urgent need to map remaining vernal pools in relatively undeveloped forested regions, such as the District Municipality of Muskoka in South-Central Ontario. This thesis aims to head-start the creation of a community-based vernal pool mapping project using LiDAR in South-Central Ontario. This goal has been broken down into two chapters with their own sub-objectives. In one chapter, we implemented a pilot study for integrating community involvement in potential vernal pool mapping across the Muskoka River Watershed (i.e., the major watershed of the District of Muskoka). We built a protocol and survey based on past vernal pool projects and studies which effectively integrated citizen involvement and also implemented novel online components (e.g., a portal) for vernal pool field-work. Our efforts were successful with positive feedback for the online components and a majority of the potential vernal pools located by our volunteers were probable vernal pools. In the other chapter, we developed two potential vernal pool mapping protocols using LiDAR based on regional characteristics of pools across the District of Muskoka in the Muskoka River Watershed and Coastal Georgian Bay. We demonstrated that the best mapping protocol for each of the two regions were associated with the protocol that was based on their respective pool characteristics. Moreover, we determined that while LiDAR can increase the accuracy of vernal pool mapping efforts, this is not always the case, especially when mapping vernal pools that occur in expansive bedrock laden regions. / Thesis / Master of Science (MSc) / Vernal pools, small forested temporary wetlands, provide essential breeding habitat for amphibians - the most threatened vertebrate group across earth. Unfortunately, most vernal pools in southern Ontario have been destroyed by human development, and there is an urgent need to map remaining vernal pools in relatively undeveloped forested regions, such as the District Municipality of Muskoka in South-Central Ontario. We implemented a pilot study for integrating community involvement in potential vernal pool mapping and developed two potential vernal pool mapping protocols using LiDAR based on regional pool characteristics. With successful integration of volunteers and mapping accuracies above 80%, we hope our findings will aid future vernal pool conservation, especially by head-starting the creation of a community-based vernal pool mapping project using LiDAR in South-Central Ontario.

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