Environmental science

The oyster mushroom, Pleurotus ostreatus, is one of the most widely cultivated edible basidiomycetes. It has gained increased attention for its economic, environmental, and medicinal properties. While a lot is known about cytokinins (CKs) and their actions at the molecular and cellular levels in plants, much less is known about the function of CKs in other kingdoms. Cytokinins, which have been detected in several fungal species, play a role in pathogenic attack against plants or during plant growth promotion by plant beneficial microbes; however, the role of CKs in fungal physiology, separate from plant associations remains largely unknown. This thesis focuses on the occurrence of fungal-derived CKs in P. ostreatus when grown in vitro as submerged or aerial mycelium. Cytokinin profiling by UHPLC-HRMS/MS revealed that P. ostreatus produces CKs and that the tRNA degradation pathway is the main source of these molecules. CK dynamics within fungal growth supported previous evidence, which suggested that tRNA degradation products have a role in the physiological development of fungi for which CKs act as fungal growth regulators. A second component of the thesis demonstrated that P. ostreatus responds to exogenous applications of aromatic and isoprenoid CKs and their effects were dependent on the dose and CK type. N6-Benzyladenine (BAP), Kinetin (KIN), N6-isopentenyladenine (iP), and trans-zeatin (tZ) bioassays revealed hormone-type responses (hormesis: biphasic response). At low doses, mycelium growth could be stimulated, whereas, at high doses only inhibitory effects were observed. This stimulation/inhibition was observed whether the measured response was an increase/decrease of aerial mycelium colony diameter, biomass accumulation or a change in mycelium morphology as compared to the controls. Results indicated there is potential to alter mycelium growth and development of P. ostreatus; thus, CKs may play the role of a "mycohormone" and may be specifically helpful for medicinal fungi by increasing growth and efficiency to produce many biologically active substances with valuable medical and environmental applications.

Author Keywords: cytokinins, fungal-derived CKs, hormesis, mycelium, mycohormone, Pleurotus ostreatus

Peatlands can be found widely across all latitudes and play a significant role in global cycles within the earth's biosphere. The anoxic conditions in peatlands promotes the accumulation of organic matter through decreased rates of decomposition and the storage of certain elements, which have received contaminant loading over the course of human existence, with significant increases occurring during the period of industrialization. We assessed global patterns of metal enrichment in peatlands in 439 cores distributed across 5 continents and 21 countries and measured 35 elements by depth increments and by peatland type. Global patterns in enrichment factors (EF's) were determined for all metals with the majority of metals being found to have a median EF < 2 indicating relatively minor enrichment. Principal component analysis indicated EF's of 6 metals (Cd, Co, Cu, Ni, Pb, and Zn), 2 metalloids (As and Sb) and Se in the upper peat horizon had similar spatial patterns among peatlands and these elements had generally the highest EF's with many cores exceeding EF >10 and some having EF values >100. Significant differences in EF's were found for these 9 "pollution" elements by peatland type and to a greater extent by geographic region, with higher EF values typically occurring in Europe and North America. Enrichment factors for most elements exhibited weak but significant positive correlations with modelled [1850 – present] S deposition. Estimated pools for the "pollution metals" within the 0 - 40 cm depth varied considerably, with median global pools in peat ranging from 12.9 mg m-2 (Sb) to 439 mg m-2 (Zn) for these 9 metals. Climate changes presents a significant risk to global peatland geochemistry due to expected changes in hydrologic regimes, resulting in potentially increased metal mobility though drought-induced peatland acidification, with historic areas previously impacted by industrial activities presenting the greatest risk of metal release to downstream receiving environments. Using a case study, we examined the impact of simulated 30-day drought on pore water chemistry at six sites in a peatland complex in Elliot Lake Ontario that were historically impacted by uranium (U) mining activities. All sites responded similarly to simulated drought with pore water pH significantly declining. The decline in pore water pH was likely due to increasing sulphate (SO42-) concentrations, which accompanied large increases in Al, Ni, Cu, Pb, Zn, and U. Dissolved organic carbon (DOC) increased, which may further enhance Al, Cu, and U mobility as these metals are strongly complexed by organic acids. Metal partitioning (Kd) values could be significantly predicted by pH and DOC although the strength of the relationship varied considerably among sites. Multiple linear regression and the inclusion of SO4-2 improved predictions, indicating that declines in pH as a result of SO4-2 and H+ production primarily governs metals and U partitioning in peatland soils. The results from both studies show that metal enrichment in global peatlands is highly variable, with northern peatlands in industrialized areas presenting the greatest risk of metal release to downstream surface waters based on expected hydrologic impacts from climate change due to historical and on-going metal and S deposition.

Author Keywords: Acidification, Climate Change, Drought, Enrichment Factors, Global, Peatlands

Mixtures of metals occur in surface waters, toxicity of which has drawn world-wide attention due to their crucial role in both ecotoxicology and regulations. The present research was undertaken to study the acute toxicity of binary mixtures of Ni, Cu, Zn, and Cd to the freshwater organism, Daphnia magna. The experimental approach included single and binary metal toxicity tests based on the 48h acute toxicity bioassay of Environment Canada. The acute toxicity of single metals followed the order of Cd > Cu > Zn > Ni. Based on the calculated 48h EC50 value of single metals, a toxic unit (TU) approach was used to combine two metals in a binary mixture, in which 1TU was equal to the 48h EC50 value of a metal in single exposure. The toxicity of binary metal mixtures to D. magna followed the order of Cu-Cd > Cu-Zn > Zn-Cd > Cu-Ni > Zn-Ni > Cd-Ni, which demonstrated three types of toxicity (i.e., less than additive, additive, and greater additive). Predictions from additivity models (including concentration addition (CA) and independent action (IA) models), a generalized linear model (GLM), and a biotic-ligand-like model (BLM-like) were compared to the bioassay results. The CA and the RA models also predicted three types of toxicity of the binary metal mixtures (i.e., less than additive, additive, and greater than additive). However, the CA model mostly overestimated the toxicity of binary mixtures. Predictions from the GLM supported the inclusion of the interaction between two metals in a mixture to predict the toxicity of binary metal mixtures. The binary metal toxicity was also predicted using a BLM-like model based on the calculated concentrations of free ionic forms of the metals, affinity constants, and toxic potency of each metal. In this model, it was hypothesized that the toxicity of metal mixture is the result of competition of metals with Ca2+ at biotic ligands, which can lead to whole-body deficiency of Ca2+ in D. magna. The BLM-like model provided the toxic potency of single metals with the following order, Cu > Cd > Zn > Ni. Although the prediction of the BLM-like model was not in good agreement with the observed toxicity of binary metal mixtures, an overestimation of risk of mixture toxicity was obtained using this model, which could be promising for use in environmental risk assessment.

Author Keywords: biotic ligand model, concentration addition, Daphnia magna, independent action, metal toxicity, modeling

Soil and tree chemistry were measured across 15 limed sites that were established 14 to 37 years ago within the Sudbury barrens in Ontario, along with two unlimed pre-treatment condition reference sites and an unlimed remnant pine forest. Soil pH and base cation (calcium (Ca), magnesium (Mg), and potassium (K)) concentrations were elevated in surface organic [FH] horizons up to 37-years post limestone treatment. Limestone in the organic horizon was evident by higher Ca/Sr ratios (a good marker of dolomite) in younger sites. Base cation mass budgets were generally unable to account for the mass of added Ca and Mg. Sudbury is characterized by widespread metal contamination. Metal (copper (Cu), nickel (Ni), and lead (Pb)) concentrations were generally greatest within the FH horizon and unrelated to stand age. Copper and Ni concentrations in soil generally decreased with distance from the nearest smelter. Metal partitioning (Kd) in soil was most influenced by soil pH rather than organic matter suggesting that as liming effects fade over time metal availability may increase.

Author Keywords: Afforestation, Degraded, Limestone, nutrient, Space-for-time, Sudbury

Lake of the Woods (LOW) is a large international waterbody which suffers from frequent and widespread algae blooms. Previous studies have highlighted the importance of the lake's largest tributary, the Rainy River (RR) and its significance in total phosphorus (TP) delivery to the LOW. Unfortunately, little is known about TP contributions from the RR and its tributaries within the Canadian portion of the watershed. This thesis examines patterns and sources of TP from four tributaries on the Canadian side of the lower RR region, two of which are predominantly natural, and two that are predominantly agricultural. Relationships between water quality parameters, land use and geologic characteristics were observed over a complete hydrologic year (Oct 1, 2018 - Sept 31, 2019), and through an intensive sampling campaign using a nested watershed approach during the spring high flow and summer low flow periods. Results revealed that TP and total suspended sediment (TSS) concentrations (>100 µg/L and >20 mg/L respectively), and loads (>20 kg/km2 and >3500 kg/km2, respectively), were greater at agricultural sites compared with natural sites (<65 µg/L TP and <15 mg/L TSS concentration, and <20 kg/km2 TP and <4000 kg/km2 TSS export). Total P, TSS, Fe, and Al were significantly positively correlated (R2= 0.26-0.59; p<0.05) and intensive sampling revealed that these relationships were strongest during the spring and at the agricultural sites (R2= 0.73-0.98; p<0.05). In contrast, the summer intensive sampling revealed that TP and redox sensitive Fe were significantly correlated (R2= 0.72; p<0.005), whereas redox insensitive Al and TSS were not, suggesting TP may be sourced via redox processes in the summer due to favourable hydrologic conditions. This was observed not only at sites with high wetland influence, but also at sites with more agricultural presence suggesting that redox sourced TP may also originate from mineral stream bed sediment during low flow periods. This research suggested two primary TP sources in the lower RR region: erosion in the spring, and redox processes (internal release) in the summer. It is recommended that intensive monitoring continue in Canada, and further research be conducted to fully understand the significance of internal P release in the tributaries.

Author Keywords: erosion, land use, nutrients, particulates, redox, water quality

This thesis presents an exploration of the requirements for and development of a software tool to calculate Certified Emission Reduction (CERs) credits for afforestation and reforestation projects conducted under the Clean Development Mechanism (CDM). We examine the relevant methodologies and tools to determine what is required to create a software package that can support a wide variety of projects involving a large variety of data and computations. During the requirements gathering, it was determined that the software package developed would need to support the ability to enter and edit equations at runtime. To create the software we used Java for the programming language, an H2 database to store our data, and an XML file to store our configuration settings. Through these choices, we can build a cross-platform software solution for the purpose outlined above. The end result is a versatile software tool through which users can create and customize projects to meet their unique needs as well as utilize the features provided to streamline the management of their CDM projects.

Author Keywords: Carbon Emissions, Climate Change, Forests, Java, UNFCCC, XML

In this study we seek to better understand the potential effects of short-term (5-year) N fertilization on jack pine forest biogeochemistry, vascular plant community composition and to project a temporal endpoint of nitrogen leaching below the major rooting zone. Aqueous ammonium nitrate (NH4NO3) was applied above the forest canopy across five treatment plots (20 x 80 m) four times annually. The experimental deposition gradient followed those known for localized areas around the major open pit operations at 0, 5, 10, 15, 20 and 25 kg N ha-1 yr-1 over a five-year period (2011 – 2015). Nitrate recovery in throughfall was significantly higher than NH4+ (p < 0.05), indicating canopy NH4+ immobilization. There was a strong treatment effect (p < 0.05) of N on the epiphytic lichen thalli concentrations of Hypogymnia physodes and Evernia mesomorpha after five years. The canopy appeared to approach saturation at the highest deposition load (25 kg N ha-1 yr-1) during the fifth year of N additions as most N added above the canopy was accounted for in throughfall and stemflow. The non-vascular (lichen and moss) vegetation pool above the forest floor was the largest receptor of N as cryptogam foliar and thalli N concentrations showed a significant treatment effect (p < 0.05). Nitrogen in decomposing litter (25 kg N ha-1 yr-1) remained immobilized after five years, while treatments ≤ 20 kg N ha-1 yr-1 started to mobilize. Understory vascular plant cover expansion was muted when deposition was ≥ 10 kg N ha-1 yr-1. Finally, modeling suggests the forest may not leach N below the rooting zone until around 50 years after chronic addition begin (25 kg N ha-1 yr-1). The modeling results are consistent with empirical data from a high exposure (~20 - 25 kg N ha-1 yr-1) jack pine site approximately 12 km west of the experimental site that has not yet experienced N leaching.

Author Keywords: Biogeochemistry, Canopy, Deposition, Jack Pine, Nitrogen, Understory

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

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

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