Environmental and Life Sciences

In Ontario, farmers commonly use a MZ (Maize (Zea mays L.))-SB (Soybean (Glycine max))/WW (Winter wheat (Triticum aestivum)) – CC (mixed cover crop) rotation to maximize economic benefits. This study aimed to investigate the short-term impacts of the crop rotation phases and their associated management practices in this diversified cash crop rotation on soil health and the abundance of nitrogen (N)-cycling soil microbial communities (SMCs). Additionally, the abundance of N-cycling SMCs and plant-available N in both surface (0-5 cm) and rooting zone (5-15 cm) depths were characterized in tile-drained (TD) and non-TD fields. In the present study, soils collected under the CC phase had the highest labile carbon levels (10-17% higher) and water-stable aggregates (35-50% higher) compared to the other two crop phases. Lower nitrifying (amoA) gene abundances and soil NO3--N levels were observed in the CC phase compared to the MZ and SB-WW phases, suggesting a potential for decreased nitrification in the CC phase. The presence of SB potentially influenced the soil N concentration in the subsequent WW phase likely due to the release of symbiotically fixed N in the SB-WW phase. Further, higher amoA abundances and NO3--N in the SB-WW phase imply a potential for increased nitrification in the SB-WW phase. Additionally, higher amoA/nosZI and nirS+nirK/nosZI ratios were observed in the MZ phase than in SB-WW and CC phases, suggesting a potential capacity for increased N2O emissions from the reactions mediated by N-cycling SMCs in soils planted to MZ during fall sampling days. In the TD and NTD field study, higher NO3--N levels were observed in TD-SB-WW fields at 5-15 cm vs. 0-5 cm depths, which was possibly facilitated by tile drainage. The TD-CC fields displayed higher nosZI gene abundances and lower nirS+nirK/nosZI abundance ratios, suggesting a greater potential capacity for decreased N2O emissions in soils planted to CCs during the spring sampling days. When examining changes in plant available N by soil depth, reduced downward movement of NO3- through shallow soil depths (0-15 cm depth) was observed in the CC phase compared to cash crops. This short-term study highlights the potential contribution of the CC phase, particularly within TD agricultural fields, for improving soil health and reducing potential N2O emissions. Together, these results suggest that management-associated differences in crop rotation phases have temporary effects on soil health and the abundance of SMCs. Future studies linking N-cycling SMC's potential activity and field-scale N2O fluxes will provide a better insight into the longer-term sustainability of Ontario's cash crop management systems.

Author Keywords: denitrification, maize-soybean-winter wheat- cover crop rotation, nitrification, soil depth, Sustainable agriculture, tile-drainage

In an effort to improve upon existing analytical methods, electrochemical sensors offer portable, cost-effective alternatives to traditional lab-based techniques. Recent advances in supramolecular chemistry offer a unique alternative to achieve high selectivity while also benefitting from facile scaling for mass production. Thus, by incorporating host-guest chemistry with electrochemical sensors, the development of simple and selective sensors is possible. To that extent, novel hosts and electroactive ion pairs were investigated for their ability to transduce an electrochemical signal representative of host-guest complexation. Results demonstrated that the upper rim modifications of resorcinarene hosts attenuated their affinity for electroactive probes whilst maintaining structural integrity upon extended cycling. Further work revealed that guests may be directly quantified via their complexation with electroactive hosts. The sensing method was further validated by quantification of surfactant pollutants in the Otonabee River. Through a fundamental understanding of the electrochemical behaviour of host-guest systems a general sensing platform can be developed, where hosts are interchangeable for specificity towards any desired analyte. Therefore, moving away from expensive lab-based methods and significantly reducing the barriers for biological or environmental monitoring.

Author Keywords: Electrochemistry, Ferrocene, Host-Guest, Resorcinarenes, Supramolecular Chemistry, Surfactants

The detection of ciguatoxins (CTXs) in biological samples is challenging due to their low concentrations, the presence of various congeners, and the absence of standardized methods. This study uses high resolution mass spectrometry (HRMS) with P-CTX-3B as a reference standard. The protonated molecules ([M+H]+) were most prevalent, especially when acetonitrile/water was utilized, providing enhanced sensitivity. Optimized collision energies of 15 eV for protonated molecules and flow rates of 10 µl/min enhance sensitivity and peak intensities, respectively. Acetonitrile/water (ACN/H2O) is recommended as the primary solvent for HRMS method, an aspect underexplored in existing literature. The detection of CTX-3B in fish tissue samples proved to be challenging, caused by variations in ion peak intensities and matrix effects, requiring a deeper exploration of the impact of complex matrices on CTX detection. The study emphasizes the need for a reliable internal standard to mitigate these effects and highlights the ongoing challenge of developing a rapid, simple, and sensitive detection method. The study's specific focus on the P-CTX-3B analogue significantly contributes to methodology development for this congener, serving as a foundational step in understanding and detecting CTX. Despite notable progress, the study acknowledges the absence of an ideal assay, outlining key challenges for future research on ciguatera analysis. It underscores the continuous necessity for method reevaluation, testing, and the broader goal of establishing a more clarified and rugged method for the identification of CTX in fish.

Author Keywords: Analytical Chemistry, Ciguatera Fish Poisoning, Ciguatoxin, High-Resolution Mass Spectrometry, Optimization, P-CTX-3B

Molecular and genomic tools provide a deeper understanding of the ecology and evolution of species and their capacity to adapt to changing selective pressures, where diversity is presumedly linked to higher fitness and evolutionary potential. Molecular tools can also illustrate how historical processes affect contemporary genetic variation to predict how current population trends may influence future genetic diversity. Genomic investigations increasingly extend beyond variation within host genomes to include diversity of their associated microbiomes, recognized to influence host/environment interactions and adaptation. Muskoxen (Ovibos moschatus) are iconic, Arctic herbivores of ecological, economic, and cultural significance. Demographically, most mainland muskox populations have remained stable or grown over the last century, yet the biggest herds, found on Victoria and Banks Islands (Nunavut and the Northwest Territories, Canada) have experienced recent and drastic population declines. These Arctic island population declines have been associated with warming trends leading to shifting ranges of forage biodiversity, and pathogen expansions directly associated with increased mortality. Genomic investigations have the potential to enhance understanding of these contrasting trends and the adaptive capacities of muskox to cope with rapid ecological change. In this thesis, I assess genetic, genomic, microbiome and diet diversity to better understand the ecology, and evolution of muskoxen. I found extremely low levels of genetic variation associated with population bottlenecks coinciding with major glaciation events and contemporarily low levels of gene flow among populations. Whole genome analyses identified signatures of selection between muskox populations, providing a genetic basis for the divergence of two previously proposed muskox subspecies. Significant differences in diversity, effective population size and inbreeding among subspecies suggests animals from Arctic islands and Greenland are more vulnerable to environmental change. Molecular investigations of diet and microbiome diversity reflected unique capacities of muskoxen to survive on high-fiber forage and exploit shifts in Arctic vegetation that may include continued shrubification. Overall, these data provide insight into the complex relationship between genetic diversity and changing environments, setting a foundation for expanded future investigations of muskox seeking to promote the future viability of this species.

Author Keywords: Genetic Diversity, Genome Assembly, Metabarcoding, Microsatellites, Muskox, Persistence

Cytokinins (CKs) are adenine derivative molecules that are present in all kingdoms of life. CKs are known to have a role in cell growth and development in plants. However, the role of CKs in vertebrate systems is not well understood. Frog virus 3 (FV3) is a type species of the Iridioviridae family, genus Ranavirus. FV3 is a major contributor to the amphibian population decline in North America. In this study, we demonstrate that concurrent and pretreatment of 20 µM of either N6-isopentyldenine (iP), N6-ispoentyladenosine (iPR), N6-furfurladenine/kinetin, and N6-furfurladenosine/kinetin riboside (KR) decreased FV3 replication. To understand the mechanism of inhibition, we assessed morphological changes in host cell nuclei to assess the effect of CKs on infected nuclei. Our results show that infection with FV3 and 20µM treatment of iP or iPR reduced nuclei size. These results are the first to reveal insight into the potential mechanism in which FV3 replication is inhibited by iP and iPR.  

Author Keywords: cytokinins, frog virus 3

Estimation of population abundance from samples has inherent practical challenges. Moreover, analytical methods to estimate abundance may vary in statistical assumptions and prediction uncertainties. I evaluated the performance of design-based and model-based methods to estimate Canada geese (Branta canadensis) abundance based on aerial fixed-width transect surveys in the Hudson Bay Lowlands, Canada. I evaluated Empirical Bayesian Kriging (EBK), areal interpolation and a ratio estimator on the basis of accuracy and precision using spatial point simulations. Untransformed EBK was the most accurate and precise, due in part, to its inherent handling of nonstationary distributions. The ratio estimator followed the same trends as EBK and, in some cases, had higher precision. Consideration of alternative analytical methods and their strengths and weaknesses is an important step in generating reliable information for population monitoring. Geostatistical approaches such as EBK have the benefit of providing spatially explicit mapping of abundance and reliable population estimates.

Author Keywords: Areal interpolation, Design-based inference, Empirical Bayesian Kriging, Geostatistics, Model-based inference, Ratio estimator

Northern flying squirrels (NFS) are mycophagous specialists (fungi-dominated diet) thatmay be displaced with southern flying squirrel (SFS) range expansion, thereby limiting fungal dispersal in forest communities. To understand the implications of squirrel species turnover on mycophagy, we investigated the home ranges of both flying squirrel species who are living in stable sympatry. We found no significant difference in home range sizes and identified spatial overlap between the two species. Through habitat selection ratios we found SFS were strongly selecting for deciduous-dominated habitats more than NFS. Lastly, we conducted microscopy on flying squirrel scat and found NFS were eating more fungi than SFS. We conclude that the squirrels are sharing the same habitat landscape but are finding ways to partition the habitat accordingly to allow for sympatry. SFS may contribute to the spore-dispersal cycle similarly to their northern counterpart through moderate fungus consumption and large home range sizes.

Author Keywords: diet, flying squirrels, Glaucomys, home range, mycophagy, sympatry

Climate change is a major conservation concern, especially for many cold-adapted species. The rate of warming due to climate change will likely outpace adaptive responses, and many populations will likely need to rely on phenotypic plasticity to cope with environmental warming. It is currently unclear whether plasticity in physiological responses to warming will be sufficient to offset the negative consequences of chronic environmental warming in ectotherms. I studied within-generation and transgenerational plasticity in two cold-adapted species of fishes, lake trout (Salvelinus namaycush) and brook trout (S. fontinalis), following temperature acclimation. Adults of both species were acclimated to either cold or warm temperatures and offspring were generated using a fully factorial breeding design, whereby the family thermal histories included crosses made within each temperature treatment and bidirectional crosses between temperatures. Offspring families were subdivided into two groups and acclimated to either warm or cold temperatures, so that offspring thermal experience matched or mismatched that of one or both parents. Offspring metabolic rate and critical thermal maximum during an acute thermal challenge were measured for both species. Limited transgenerational plasticity was detected in both species, but had a lesser effect than within-generationacclimation. In brook trout, the paternal contribution was greater than the maternal contribution. In lake trout, a mismatch in thermal acclimation, where the offspring were cold-acclimated but the parents warm-acclimated, resulted in elevated offspring metabolic rate without a corresponding increase in growth, suggesting that a mismatch in temperatures across generations could be detrimental to offspring. Using RNA-sequencing, transgenerational plasticity was linked to differential gene expression in the liver of lake trout offspring, in that genes were differentially expressed depending on the parental acclimation temperatures. Within-generation warm acclimation had the greatest effect on gene expression profile of offspring, with more genes differentially expressed under conditions of within-generation warm acclimation compared with transgenerational warm acclimation. Although it has been suggested that transgenerational plasticity may help to buffer the impact of warming due to climate change, my work implies that transgenerational plasticity, like within-generation plasticity, will be insufficient for these two species of cold-adapted salmonids to cope with climate change.

Author Keywords: Brook trout, Climate change, Lake trout, Phenotypic plasticity, Thermal tolerance, Transgenerational plasticity

Methylobacterium spp., a dominant and functionally conserved group of plant-associated bacteria, have long been recognized for their roles in promoting host growth, stress tolerance, and phytohormone modulation. This body of work collectively repositions Methylobacterium not only as a plant growth-promoting genus but also as a promising agent of microbiome-mediated crop protection. Across several investigations, the ecological, biochemical, and functional attributes that underpin this potential were examined, with specific focus on hormone production, compatibility with agrochemical inputs, and antifungal activity.

A comprehensive inventory of 46 Methylobacterium strains revealed widespread production of cytokinins – including highly active forms such as trans-zeatin – and variable capacities to synthesize indole-3-acetic acid. Cytokinin output increased under carbon-limiting conditions, highlighting the genus's adaptive hormonal response. Parallel investigations demonstrated that commercial glyphosate-based herbicide formulations significantly inhibited the growth of most Methylobacterium strains, whereas pure glyphosate alone showed negligible toxicity. Key findings of experiments indicate that non-active formulation components participate in the disruption of beneficial bacteria by facilitating higher intracellular glyphosate concentrations and subsequent toxic effects. This introduces a novel link between agrichemical formulation practices and the selective disruption of keystone microbial taxa.

Contrastingly, fungicide compatibility testing showed that Methylobacterium strains tolerate key fungicides such as azoxystrobin, fludioxonil, and metalaxyl-M, supporting their inclusion in integrated pest management frameworks. Subsequent functional antagonism assays further revealed that specific Methylobacterium isolates inhibit phytopathogenic Fusarium species in vitro and in planta. Notably, M. organophilum enhanced soybean seedling vigor and reduced disease severity when co-inoculated with F. graminearum by preserving the integrity of the seed coat, demonstrating protective activity with unique mechanics.

Finally, differential hormone profiling at the pathogen-antagonist interface revealed that biocontrol-effective Methylobacterium strains not only produce higher levels of auxin and salicylic acid but also induces jasmonic acid production – likely derived from Fusarium – suggesting complex cross-signalling and interference with fungal development and sensing pathways. Together, these findings advance our understanding of Methylobacterium as a keystone genus in the phytobiome, capable of contributing to both plant vigor and pathogen suppression and reinforce its relevance in the design of next-generation biocontrol strategies.

Author Keywords: agrochemical interactions, biological control, Fusarium antagonism, Methylobacterium, phytobiome, phytohormone signalling

The Wabigoon River is known for an historic mercury (Hg) pollution source, caused by a chlor-alkali facility operating in the 1960s. As legacy Hg contamination continues to cause serious adverse health effects to the local communities living in the Wabigoon River region, it is imperative to undertake additional research to understand the deposition and transport of historical mercury in this system and more importantly, its conversion into methylmercury (MMHg) which renders it bioavailable for ongoing bioaccumulation. The aim of this dissertation was to evaluate the transport and accumulation of Hg species by doing a spatial and temporal analysis of concentrations of mercury and methylmercury along the Wabigoon River, as well as assessing rates of methylation and demethylation, identifying areas of higher methylmercury production. Results show that locations downstream from the pollution source still show elevated mercury concentrations, with levels at least five times higher in water and up to 134 times higher in sediments compared to background levels. Among selected study sites, the Hydroelectric dam, the Wabigoon Rapids wetland and Clay Lake were identified to have high capacity for methylmercury production in the system, with notably Clay Lake presenting a higher potential for methylmercury accumulation due to the observed lower methylmercury demethylation rate. Furthermore, the impact of wetting and drying cycles on Hg methylation in riverbed and wetland locations within the Wabigoon River system was investigated through a laboratory simulation. Findings indicated increased susceptibility of riverbed locations to wetting and drying cycles.

Author Keywords: Demethylation, Mercury, Methylation, Methylmercury, Wabigoon River