Graduate Theses & Dissertations


Using environmental DNA (eDNA) metabarcoding to assess aquatic plant communities
Environmental DNA (eDNA) metabarcoding targets sequences with interspecific variation that can be amplified using universal primers allowing simultaneous detection of multiple species from environmental samples. I developed novel primers for three barcodes commonly used to identify plant species, and compared amplification success for aquatic plant DNA against pre-existing primers. Control eDNA samples of 45 plant species showed that species-level identification was highest for novel matK and preexisting ITS2 primers (42% each); remaining primers each identified between 24% and 33% of species. Novel matK, rbcL, and pre-existing ITS2 primers combined identified 88% of aquatic species. The novel matK primers identified the largest number of species from eDNA collected from the Black River, Ontario; 21 aquatic plant species were identified using all primers. This study showed that eDNA metabarcoding allows for simultaneous detection of aquatic plants including invasive species and species-at-risk, thereby providing a biodiversity assessment tool with a variety of applications. Author Keywords: aquatic plants, biodiversity, bioinformatics, environmental DNA (eDNA), high-throughput sequencing, metabarcoding
Hydrological and Flooding Effects on Stream Nutrient Levels
Stream solutes are strongly linked to hydrology, and as such, we sought to better understand how hydrology, particularly flooding, influences nitrogen (N) and phosphorus (P) levels. We used a long-term dataset of monthly water quality samples for many Ontario, Canada, catchments to assess the effects of landscape variables, such as land use and physiography, on the export of nutrients during floods, and to characterize overall concentration-discharge patterns. In general, we found that landscape variables could partially explain the export variation in flood waters, but that the importance of specific variables depended on flood characteristics. We also found that overall concentration-discharge relationships for N and P C were positive, but non-linear, with greater concentrations on the rising limb of the hydrograph depending on the nutrient. With these results, we have identified general patterns between nutrients and hydrology, which will be helpful for managing the ecological effects of flooding. Author Keywords: C-Q relationships, Discharge, Export, Flooding, Nutrients, Thresholds
Discontinuities in stream networks
The network composition hypothesis (NCH) suggests that i) large confluence symmetry ratios (drainage area of the tributary relative to the mainstem) and ii) landscape differences (differences in landscape characteristics between the mainstem and tributary drainages) lead to greater ecological changes below confluences. As a test of the NCH, 34 confluences were sampled in southern Ontario to examine the effects of these two factors on benthic invertebrate communities to infer the degree of ecological change at confluences. Given the typology of streams surveyed, there was subtle evidence that benthic invertebrate communities below confluences changed as a function of confluence symmetry ratio and landscape differences. This indicates that abrupt changes in stream networks are not as common as theory may suggest. Further support for the network composition hypothesis may be found by examining a wider range of stream types and examining single-species responses. Author Keywords: benthic invertebrates, community similarity, landscape characteristics, stream networks, tributary
wind tunnel and field evaluation of the efficacy of various dust suppressants
A series of experiments was designed to assess the relative efficacy of various dust suppressants to suppress PM10 emissions from nepheline syenite tailings. The experiments were conducted in the Trent University Environmental Wind Tunnel, Peterborough, Ontario, and on the tailings ponds at the Unimin Ltd Nephton mine near Havelock, Ontario. Treated surfaces were subjected to particle-free airflow, abrasion with blown sand particles, particle-free airflow after physical disturbance, and were measured independently using a pin penetrometer. In the particle-free wind tunnel tests, three of the surfaces performed well, and PM10 emissions scaled inversely with crust strength. Light bombardment of each surface by saltating sand grains resulted in PM10 emission rates two orders of magnitude higher. All treated surfaces emitted significantly more PM10 after physical disturbance in both the laboratory and field research. The results suggest that the site conditions, inclusive of the potential for dust advection and resuspension, must be taken into account when considering the use of a commercial dust suppressant. Author Keywords: dust suppression, field testing, mine tailings, wind tunnel experiment
Temperature effects on the routine metabolic rates of brook trout (Salvelinus fontinalis) eggs, alevin and fry
Early developmental stages of cold-adapted ectotherms such as brook trout (Salvelinus fontinalis) are at risk of mortality with increasing water temperatures because of their sensitivity to changes in their environment. I studied the mass and routine metabolic rate (RMR) of wild-origin brook trout eggs, alevin and young fry reared at normal (5°C) and elevated (9°C) temperatures for the duration of the study or at mismatched temperatures. This setup determined if preconditioning acclimation for one temperature benefits or hinders the organism later in life. Three levels of biological organization (ancestry, population, family) were studied using Akaike’s Information Criterion (AIC) to identify models that best accounted for variation in the data. Family, mass and temperature were most important in predicting body mass and mass-adjusted RMR, although population and ancestral-level differences were also detected at some life stages. Strong variation in body mass and mass-adjusted RMR among families may indicate adaptive potential within brook trout populations to respond to increases in water temperature with climate change. Author Keywords: Acclimation, AIC, Brook trout (Salvelinus fontinalis), Environmental matching, Routine metabolic rate, Temperature
Effect of the neonicotinoid imidacloprid on embryogenesis and anuran survivorship in frog virus 3 infected tadpoles
Exposure of pre-metamorphic amphibians to neonicotinoid insecticides may be contributing to the global decline in amphibian populations. In this study, anuran embryos and tadpoles of the African clawed frog (Xenopus laevis) and the North American leopard frog (Lithobates pipiens) were used to determine the effects of embryonic exposure to neonicotinoids. In addition, Xenopus was used to determine if prolonged exposure to neonicotinoids influenced tadpole sensitivity to frog virus 3 (FV3). Exposure of anuran embryos to concentrations of the neonicotinoid insecticide, imidacloprid, ranging from 1 -20 ppm induced a concentration dependent increase in malformations of the retina in Xenopus embryos. However, similar responses were not observed with embryos of leopard frogs. Exposure of Xenopus tadpoles to 500 ppb concentration of imidacloprid followed by challenge with FV3 showed that pesticide exposure unexpectedly decreased the rates of mortality, although total mortalities by the end of the experiment were not significantly different from controls. This unexpected observation may be attributed to a reduced inflammatory response induced by exposure to imidacloprid. Despite the low acute toxicity of neonicotinoid insecticides to vertebrates, these studies indicate that exposure to this class of insecticides causes sublethal effects in anuran species during early life stages. Author Keywords: embryogenesis, Lithobates pipiens, neonicotinoid, ranavirus, tadpole, Xenopus laevis
Biology and Management of Stratiotes Aloides in the Trent River, Ontario
Invasive aquatic plants can create negative ecological, economic and social impacts when they displace local vegetation, interfere with shipping and navigation and inhibit water-based recreational activities. In 2008, the first North American occurrence of the invasive plant Stratiotes aloides (Water soldier) was identified in the Trent River, Ontario. This research measured offset photosynthesis and turion germination to determine the light compensation point (5.2-5.4m) and maximum depth of colonization (4-6m) for S. aloides propagules using in situ incubations and controlled growth experiments. The effects of spring and fall chemical (Diquat) and physical (hand raking) treatments on S. aloides biomass, local macrophyte recovery and community dynamics in the Trent River were also measured. The target of a 75% minimum reduction in S. aloides biomass was not attained using any of the treatment methods and no perceivable recovery of the local plant community was observed. Significant S. aloides regrowth was recorded for both treatment methods regardless of application timing. Author Keywords: herbicide, invasive species, macrophyte, photosynthesis, propagule
impact of selection harvesting on soil properties and understory vegetation in canopy gaps and skid roads in central Ontario
Tree harvesting alters nutrient cycling and removes nutrients held in biomass, and as a result nutrient availability may be reduced, particularly in naturally oligotrophic ecosystems. Selection harvesting is a silvicultural technique limited to tolerant hardwood forests where individual or small groups of trees are removed creating a “gap” in the forest canopy. In order for harvesting machinery to gain access to these individual trees, trees are felled to create pathways, known as skid roads. The objective of this study was to characterize differences in soil chemical and physical properties in gaps, skid roads and uncut areas following selection harvesting in central Ontario as well as documenting differences in the understory vegetation community and sugar maple (Acer saccharum) seedlings chemical composition post harvest. First year seedlings were collected for elemental analysis from unharvested areas, canopy gaps, and skid roads in 2014, eight months after harvesting. In 2015, first and second year sugar maple seedlings were collected. Soil bulk density and water infiltration were measured in the three areas of the catchment as well as soil moisture, organic matter content, exchangeable base cations, and net nitrification. Seedlings in the disturbed sites had lower concentrations of Mg, K, P, and N compared with unharvested sites and soil nitrification was significantly lower in the skid roads. Water infiltration rates in the gap and skid roads were slower than the control and concentrations of metals (e.g. Fe, Al, Ca) and litter mass increased in litter bags deployed over 335 days, likely reflecting an increase in soil erosion in the skid roads. Understory vegetation was markedly different amongst sites, particularly the dominance of Carex spp. in the skid roads. The sustainability of industrial logging is dependent on successful tree regeneration, however, increased soil compaction, establishment and growth of grasses and shrubs, as well as low nutrient concentrations in seedlings may ultimately restrict forest succession. Author Keywords: Canadian Shield, nitrification, selection harvesting, soil compaction, sugar maple seedling, understory vegetation
Evaluating Environmental DNA (eDNA) Detection of Invasive Water Soldier (Stratiotes Aloides)
In 2008, the first North American water soldier (Stratiotes aloides) population was discovered in the Trent River, Ontario. Water soldier is an invasive aquatic plant with sharp, serrated leaves that has the potential to spread rapidly through dispersed vegetative fragments. Although it is too late to prevent water soldier establishment in the Trent River, its local distribution remains limited. In this study, environmental DNA (eDNA) was explored as a potential tool for early detection of water soldier. Species-specific markers were designed from chloroplast DNA regions matK and rbcL, and a qPCR assay with rbcL primers yielded the most sensitive detection of water soldier eDNA. Positive detections were obtained from six of 40 sampling locations, of which five were collected in Seymour Lake, an area with large patches of water soldier. As water soldier plants were known to be present at these sites, high eDNA concentrations were expected. The sixth positive detection from Trent Lock 5 (50 km downstream of Lake Seymour) was unexpected as it was obtained at a site with no water soldier sightings. This is one of the first studies to demonstrate the effectiveness of eDNA detection from aquatic plants. Author Keywords: aquatic plant, eDNA, environmental DNA, invasive species, Stratiotes aloides, water soldier
effects of in-stream woody debris from selective timber harvest on nutrient pools and dynamics within Precambrian Shield streams
Timber harvest can influence the rate of transfer of organic matter from the terrestrial catchment to streams, which may have both direct and indirect effects on in-stream nutrient pools and dynamics. In the interest of developing sustainable forestry practices, the continued study of the effects of forestry on nutrient dynamics in aquatic systems is paramount, particularly in sensitive nutrient-poor oligotrophic systems. The goal of this study was to investigate the impacts of harvest-related woody debris on stream nutrient status in streams located in the Canadian Shield region of south-central Ontario. Surveys showed greater large (> 10 cm) and small (< 10 cm) woody debris dry masses and associated nutrient pools in streams located in recently (2013) selectively harvested catchments, when compared with catchments not harvested for at least 20 years. Experimental releases of flagging tape underlined the importance of woody debris as a mechanism of coarse particulate organic matter (CPOM) retention. Sediment surveys showed a significant exponential decline in both OM content and nutrients associated with coarse sediment with distance upstream from debris dams. Laboratory leaching experiments suggest that fresh woody debris may be an important short-term source of water-soluble nutrients, particularly phosphorus and potassium. This study suggests that woody debris from timber harvest is both a direct and indirect source of nutrients, as trapped wood and leaves that accumulate behind debris dams can augment stream nutrient export over long time periods. Author Keywords: nutrient leaching, nutrient pools, organic matter retention, selection harvest, southern Ontario, woody debris
Chemical characterization of dissolved organic matter in relation with hydrography in the Arctic Ocean
In this thesis, water mass distribution of dissolved organic matter (DOM) characteristics (i.e. molecular weight, fluorescent components, thiols and humic substances concentration) was observed in the Arctic Ocean. For the first time, DOM molecular weight (MW) in Beaufort Sea was assessed using asymmetrical flow field-flow fractionation, as well as the first monitoring of thiols and humic substances (HS) using cathodic stripping voltammetry (CSV) in the Arctic Ocean. Based on fluorescence property, DOM characterization was carried out using parallel factor analysis – excitation-emission matrices. Pacific winter waters in the Canada Basin showed higher MW DOM associated with higher fluorescence intensity. High HS was associated with the Arctic outflow waters in top 300 m of the Canadian Arctic Archipelago. Interestingly, maximum thiol concentration was associated with the subsurface chlorophyll-a maximum at most sites, but not universal along the study area. Comparable distributions of CSV-based HS and humic-like fluorescent components suggest similar sources/ processes in the Arctic Ocean. The findings in this thesis suggested DOM characteristics could be used as fingerprints in tracing water masses in the Arctic Ocean. Author Keywords: Asymmetrical flow field-flow fractionation, Cathodic stripping voltammetry, DOM, Metal-binding ligands, Molecular weight, PARAFAC-EEMs
Responses of Primary Producers and Grazers to Silver Nanoparticle Exposure
The increasing production and use of silver nanoparticles (AgNPs) raise concerns on environmental exposure and impact. A large scale in situ enclosure study was conducted at the Experimental Lakes Area to determine the effect of AgNPs on natural phytoplankton and zooplankton communities. This study investigated AgNPs of varying concentrations (4, 16 and 64 μg/L), dosing regimens (chronic vs. pulse), and capping agents (poly-vinyl pyrrolidone vs. citrate). Phytoplankton communities were influenced only by the natural limnological properties of the system signifying tolerance to AgNPs. Zooplankton community structure significantly changed with AgNP concentration and dosing regimen indicating AgNP sensitivity. A microcosm study investigating the effect of AgNPs and phosphorus-dosed periphyton before and after grazing by two benthic invertebrate species (snails and caddisfly larvae) showed reduced periphyton stoichiometry with AgNP exposure. Grazers foraged less on silver dosed periphyton indicating a preferential choice in food quality. Phosphorus reduced the detrimental effects of AgNPs across all conditions. These studies verify the need for in situ experimental designs to fully investigate the effects of AgNPs and their interaction with environmental factors, multiple species assemblages, and across trophic levels. Author Keywords: benthic invertebrate, Experimental Lakes Area, periphyton, phytoplankton, silver nanoparticles, zooplankton


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