Biology

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Open Access
Tethered To Telomeres: Characterizing The Form And Dynamic Elasticity Of Tethers In Anaphase Crane-Fly Spermatocytes
(2025-04-10) Economopoulos, Demetra; Connor, Steven
Elastic tethers connect sister chromosomes during anaphase in a broad range of animal cells, likely aiding in communication and coordination of movement. Calyculin A prevents dephosphorylation of tethers by protein phosphatase 1 (PP1) and, consequently, causes them to remain elastic throughout anaphase, resulting in backward movement. In my thesis experiments, I further explore the composition and behaviour of tethers. I expand upon existing research with Calyculin A, elucidating an optimal working concentration of 10 nM for investigating the reversibility of this drug on backward movement when rinsed out at various stages. I show that PP1 is critical during the onset of anaphase and that backward movement is not readily reversible. I also support the existing notion that tethers are made of titin, disproving the hypothesis that tethers are made of DNA. These contributions further enhance our understanding of tether composition and regulation during a central mechanistic phase of cell division.
Open Access
Finding Common Ground: Methods For Sustaining Citizen Science Engagement That Increase Indigenous Plant Biodiversity In Southwestern Ontario
(2025-04-10) Bava, Julia Maria; Bazely, Dawn R. A.
Nature-based solutions to address biodiversity loss require supports that reach beyond government grants. The Conservation Impact Bond (CIB) developed by Southwestern Ontario regional conservation charity, Carolinian Canada Coalition, is an example of a novel financial tool to incentivize biodiversity conservation by supporting citizen science. My research evaluated Carolinian Canada’s In The Zone Tracker and its allied programs. A systematic review of the literature about Canadian citizen science projects provided context. (1) The ITZ Program reversed plant biodiversity loss at a local level through planting projects that generated a self-reported increase in native species. (2) Information about citizen science projects was difficult to discover. Academic research into citizen science projects published in peer-reviewed literature creates a more permanent record than web-based, grey literature. (3) Citizen science projects do not necessarily improve science literacy. Rather, the ITZ tracker helped people to find common values and make positive, evidence-informed differences in their communities.
Open Access
Effects And Interactions Of Antibiotic, Butyrate, And Temperature On Sailfin Molly (Poecilia Latipinna) Metabolism And Gut Microbiota
(2025-04-10) Trovato, James; Bucking, Carol
Understanding the complex interactions between environmental factors, dietary supplements, and microbiome dynamics is crucial for optimizing fish health and growth, particularly in aquaculture settings. This study examines the effects of antibiotic, butyrate, and temperature treatments on the metabolism and gut microbiota of sailfin molly (Poecilia latipinna) alone and in combination. The results indicate that butyrate successfully restored the growth rates due to antibiotic exposure but had little effect when supplied to controls. Interestingly, while elevated temperature has been shown to promote growth through increased metabolic rates growth was not observed, possibly due to stress. Notably, there was a decrease in diversity in the microbiome because of antibiotic treatment. Further, both antibiotics and increased temperature lead to a decrease in the predicted functional capabilities of bacteria related to metabolism and health pathways after using PICRUSt analyses. While butyrate ameliorated the impacts on growth and metabolism caused by antibiotics, the microbiome was mostly unaffected, indicating that butyrate exerts its effects through the host directly. These findings suggest that butyrate could mitigate some negative impacts of antibiotics on fish growth and gut health as well as highlight some of the concerning effects of increasing environmental temperatures and antibiotic use. These insights may help develop strategies in fish population management and aquaculture practices, particularly those involving antibiotics and butyrate supplementation.
Open Access
Understanding Bumble Bee Community Changes And Floral Use In Southern Ontario.
(2025-04-10) Kerekes, Taylor; Colla, Sheila
Multiple wild bee species, including bumble bees, are in global decline due to habitat loss, reduced floral resources, and competition with managed honey bees. Monitoring previously surveyed sites is essential to assess ongoing impacts on bumble bee populations and identify trends over time. This research revisited areas in southern Ontario to examine changes in bumble bee abundance, diversity, and floral use (Chapter 1) and explored potential competition for floral resources between bumble bees and honey bees (Chapter 2). Results indicate that while some bumble bee species have increased over the last 50 years, many have declined, leading to an overall loss of diversity. Findings also reveal significant niche overlap between bumble bees and honey bees, suggesting competition for floral resources. This work informs conservation strategies, identifies species requiring focused efforts, and emphasizes the importance of maintaining diverse floral resources to support bumble bee populations.
Open Access
The Relative Importance Of Microclimatic Heterogeneity On Plant And Animal Communities In Deserts Of Southern California, U.S.A
(2025-04-10) Ghaziantafrishi, Nargol; MacDonald, Suzanne
This dissertation examines the intricate relationships between facilitation, environmental heterogeneity, and artificial habitat constructions in arid ecosystems, focusing on plant-animal and plant-plant interactions. By leveraging ecological principles, it explores the role of foundation plant species like Ephedra californica and Larrea tridentata in fostering ecosystem resilience and biodiversity under adverse conditions due to climate change, land use, and desertification. The study starts with an overview of facilitation theory, highlighting its importance in mitigating the negative effects of abiotic stressors in arid and semi-arid regions. It highlights the Stress Gradient Hypothesis (SGH) as a framework for understanding how competitive interactions can shift towards facilitation under harsh conditions, thus promoting the survival of diverse flora and fauna. The dissertation then delves into environmental heterogeneity, explaining its complex influence on ecosystem dynamics, species composition, and microclimatic regulation. Chapter 2 provides a detailed analysis of camera trapping techniques, offering optimal sampling design strategies for accurately estimating vertebrate abundance and richness. Chapter 3 examines the effects of artificial canopies on microclimate regulation. This chapter evaluates various eco-friendly fabrics, such as natural burlap, cotton, and nursery seedling fabric, in buffering microclimate through lab experiments measuring understory temperature, relative humidity, and radiation. Chapters 4 and 5 investigate how climatic variations and vegetation structure contribute to spatial and environmental heterogeneity, shaping flora and fauna diversity and abundance. A key aspect of the study is evaluating the effectiveness of artificial habitat construction as a conservation and restoration strategy in arid landscapes. Field experiments assess the microclimatic buffering capabilities of artificial shelters compared to natural vegetation in supporting vertebrate populations amid desertification in the Southwestern U.S.A. Overall, this dissertation enhances our understanding of the complex interplay between facilitation, environmental heterogeneity, and habitat modification in arid ecosystems. By uncovering the mechanisms behind these ecological processes, it provides valuable insights for conservation, management, and restoration efforts in regions vulnerable to anthropogenic disturbances and climate change.
Open Access
Examining The Role Of Lysyl Oxidase Enzymes In Atrial Fibrillation Induced By Volume Overload
(2025-04-10) Sherrard, Dana Evelyn; Backx, Peter
Atrial fibrillation (AF), the most prevalent cardiac arrhythmia, is linked to atrial electrical and structural remodelling. AF is associated with fibrosis; however, in models imposing atrial stretch, this occurs without changes in collagen transcription, suggesting fibrosis results from collagen stabilization by the Lysyl Oxidases (LOX) family of enzymes. To investigate this relationship, we established a murine aortic regurgitation (AR) model, which induces atrial stretch. AR mice exhibited increased AF susceptibility, decreased atrial refractoriness (AERP), elevated left atria (LA) fibrosis, and macrophage infiltration 2-fold compared to sham mice. LOXL2 inhibitor PXS-5382 reduced AF inducibility, shortened AF duration and attenuated fibrosis without affecting AERP. In contrast, the broader LOX inhibitor PXS-5505 prevented fibrosis without reducing AF inducibility in AR mice but exacerbated AF and fibrosis in sham mice. These results highlight LOXL2's critical role in atrial structural remodelling and AR-induced AF, suggesting it may be a promising therapeutic target for AF.
Open Access
Estimating Broad-Sense Heritability Of The Gut Microbiome In Western Honey Bees (Apis Mellifera)
(2025-04-10) Subramanian, Aishwarya; Zayed, Amro
The honey bee gut microbiome, crucial for bee health, may be shaped by both environmental and genetic factors. While differences in microbiomes exist between honey bee subspecies, it is unclear if genetic variation within a population influences this variation. This study estimated the broad-sense heritability of gut microbiome traits to explore patriline effects. We analyzed 500 workers from two colonies using 16S rRNA sequencing for microbiome characterization and genotyped them at 11 microsatellite loci. Results showed that patriline genetics marginally influenced the relative abundance of dominant microbiome taxa (18% to 24%) but did not significantly impact presence/absence of bacterial taxa or alpha and beta diversity. These findings suggest that environmental factors play a more substantial role in shaping the honey bee gut microbiome, indicating that microbiome-based interventions could benefit honey bees across populations.
Open Access
Elucidating Anthropogenic Impacts On Water Quality Through Spatial Heterogeneity In The Laurentian Great Lakes And The Global Fast Fashion Industry
(2025-04-10) Bailey, Kerrice Kesaynoi; Sharma, Sapna
Freshwater resources are vital for human survival including for consumption, transportation, and production of goods. We explore the impact of anthropogenic activity on water quality through: 1) the spatial heterogeneity in the northern nearshore regions of the Canadian Great Lakes and 2) the state of fast fashion in the top garment producing countries: China, Bangladesh, Vietnam, Turkey, India, and Indonesia. Water quality conditions varied spatially from oligotrophic conditions in Lake Superior to eutrophic conditions with elevated concentrations of nutrients and chlorophyll-a in Lake Erie. Further, we found that policy, sustainability and wastewater are key research areas in the top garment-producing countries. However, there was variation in the number of publications, terminology, knowledge gaps, and barriers that impede more sustainable garment production in the top garment-producing countries. We suggest investments in management and education to protect and use our freshwater resources more sustainably.
Open Access
The Impact Of Scene Context On Visual Object Recognition: Comparing Humans And Monkeys
(2025-04-10) Zafer, Anaa Salim; Kar, Kohitij
During natural vision, we rarely see objects on their own but rather within rich and complex environments. Understanding how the brain recognizes objects in these natural scenes by using contextual information remains a significant challenge. To explore neural mechanisms similar to human visual processing, it is crucial to have an animal model that behaves like humans. This way, the neural mechanisms we identify can offer insights relevant to the human brain. In this study, we investigated whether rhesus macaques could serve as a model for human context-driven object recognition. We did this by measuring their ability to identify visual objects with varying amounts, quality, and congruency of contextual cues. Our behavioral metrics showed remarkably similar context-dependent patterns between humans and monkeys. However, neural responses in the inferior temporal (IT) cortex of monkeys, who were not explicitly trained to recognize objects in context, could only partly explain this cross-species similarity. The behavioral similarities that were not explained by context-naive neural data highlight significant gaps in our understanding. Our findings reveal a fascinating alignment between human and monkey visual object processing that cannot be fully explained by neural activity in a key visual region.
Open Access
Investigating Pannexin1a-Mediated Mechanisms Of Pain And Neuroinflammation Using Zebrafish
(2025-04-10) Jeyarajah, Darren; Zoidl, George
This thesis explores the role of Panx1a in modulating pain and neuroinflammatory responses in zebrafish (Danio rerio). Nociception was induced using acetic acid (AA) treatments. Behavioral assays conducted on Panx1a knockout (KO) zebrafish larvae demonstrate significant alterations in response to acetic acid (AA)-induced pain. Pharmacological interventions using probenecid, a Panx1 inhibitor, and ibuprofen, a cyclooxygenase (COX) inhibitor, reveal their potential in modulating pain behaviors and rescuing nociceptive deficits. Furthermore, molecular analyses employing quantitative polymerase chain reaction (qPCR) and RNA sequencing (RNA-seq) elucidate the regulatory impact of Panx1a on gene expression related to nociception, neuroinflammation, and synaptic plasticity. In summary, this thesis provides evidence of Panx1a's involvement in pain and neuroinflammation, proposing zebrafish as a viable model for studying nociception.
Open Access
The Contribution Of Stereoposis And Motion Parallax To Depth Segmentation
(2025-04-10) Neagu, Teodora; Wilcox, Laurie
Studies comparing how stereopsis and motion parallax inform depth yield mixed results, with some showing poorer performance from motion parallax. We propose that this stems from use of depth magnitude tasks, which bias observers towards using stereopsis. In Experiment 1, we used a depth segmentation task to evaluate performance of stereopsis and self-generated motion parallax individually and in combination. Observers performed equally well with individual cues but appeared to rely on stereopsis when both were available. Follow-up experiments showed that increasing the range of self-motion did not improve performance, but the use of object motion did. Subsequently, we replicated Experiment 1 using object motion with experts and non-experts and found segmentation from object motion was always better than stereopsis, and non-experts needed more stereopsis than experts to perform the task. Thus, we suggest that the ability to use depth information depends on several factors including task demands and observers’ experience.
Open Access
Developing A Non-Human Primate Model To Dissect The Neural Mechanism Of Facial Emotion Processing
(2025-04-10) Taghian Alamooti, Shirin; Kar, Kohitij
Facial emotion recognition is a cornerstone of social cognition, vital for interpreting social cues and fostering communication. Despite extensive research in human subjects, the neural mechanisms underlying this process remain incompletely understood. This thesis investigates these mechanisms using a non-human primate model to provide deeper insights into the neural circuitry involved in facial emotion processing. We embarked on a comparative analysis of facial emotion recognition between humans and rhesus macaques. Using a carefully curated set of facial expression images from the Montreal Set of Facial Displays of Emotion (MSFDE), we designed a series of binary emotion discrimination tasks. Our innovative approach involved detailed behavioral metrics that revealed significant parallels in emotion recognition patterns between the two species. These findings highlight the macaques’ potential as a robust model for studying human-like facial emotion recognition. Building on these behavioral insights, the second phase of our research delved into the neural underpinnings of this cognitive process. We conducted large-scale, chronic multi-electrode recordings in the inferior temporal (IT) cortex of rhesus macaques. By mapping the neural activity associated with the classification of different facial emotions, we uncovered specific neural markers that correlate strongly with behavioral performance. These neural signatures provide compelling evidence for the role of the IT cortex in processing complex emotional cues. Our findings bridge the gap between behavioral and neural perspectives on facial emotion recognition, offering a comprehensive understanding of the underlying mechanisms. This research not only underscores the evolutionary continuity of social cognition across primate species but also sets the stage for future explorations into the neural basis of emotion processing. The integration of behavioral analysis with advanced neural recording techniques presents a powerful framework for advancing our knowledge of social cognition and its disorders.
Open Access
Identifying the Ecological Interactions of Invasive Lymantria Dispar Dispar and Agrilus Planipennis Using Metabarcoding Techniques
(2025-04-10) Borsato, Nicole Dorothy; Clare, Elizabeth
The objectives of this thesis were to develop a standardized metabarcoding protocol to identify ecological interactions between insects and other taxa, as well as determine the impact of ecological correlates on the detection of these interactions. In Chapter 2, I evaluate target loci for the identification of Lymantria dispar dispar (spongy moth) and Agrilus planipennis (emerald ash borer) ecological interactions and develop a standardized protocol for the analysis of these interactions. I demonstrate that my metabarcoding protocol can be used to identify a wide diversity of interactions, including those with animals, plants, bacteria, and fungi. In Chapter 3, I assess how ecological factors (e.g., life stage, collection location, etc.) influence the presence or absence of ecological interactions in emerald ash borers using random forest models. I found that interactions were more likely to be detected in pupal specimens and in samples collected earlier in the year.
Open Access
The Characterization of Taz Protein-Protein Interactions in Myogenic Cells
(2025-04-10) MacKeracher, Anastasia Elizabeth; McDermott, John
The Hippo signaling pathway is known to regulate cell proliferation, differentiation, and apoptosis. Recent studies identified the Hippo signaling target protein TAZ as a repressor of myogenic differentiation. Since TAZ lacks inherent DNA binding ability, its activity is dependent on its protein interactions. Using GFP-nanoTrap affinity purification and LC-MS/MS, we documented TAZ protein interactions in muscle cells, including the pro-myogenic methyltransferase CARM1. Investigation of the TAZ and CARM1 interaction revealed that CARM1 represses TAZ transcriptional activation, promoting TAZ Ser89 phosphorylation and cytoplasmic sequestration. The TAZ/CARM1 interaction was further investigated in embryonic rhabdomyosarcoma (RD) cells, where Hippo signaling increased compared to control C2C12 myoblasts. Further, mass spectrometry results suggest that TAZ is a substrate of CARM1 methylation, with methylation at Arg 77 enhancing TAZ Ser89 phosphorylation. These findings highlight the potential role of CARM1 in regulating TAZ function in myogenic cells.
Open Access
Investigating the Role of the E3 Ubiquitin Ligase HUWE1 in DNA Repair: Implications in Cancer Therapy
(2025-04-10) Martinho, Daniel Filipe Cruz; Sheng, Yi
The ubiquitin E3 ligase HUWE1 (HECT, UBA, and WWE containing protein 1) plays a crucial role in various cellular processes including DNA damage repair and has been implicated in ovarian cancer. This study therefore used cell-based assays to characterize the impact of HUWE1 depletion on DNA damage repair outcomes in response to different stressors. Since combinations of DNA-damaging agents with poly(ADP-ribose) polymerase inhibitors are being explored to improve ovarian cancer treatment, the impact of HUWE1 on DNA damage and survival following these combination treatments was also investigated. It was demonstrated that HUWE1 depletion increased cellular DNA damage and delayed DNA repair in response to treatment with various DNA damaging agents. HUWE1 depletion increased the synergistic effect of combination treatments with poly(ADP-ribose) polymerase inhibitors and DNA damaging agents on DNA damage and cell survival. The WWE domain of HUWE1 is known to interact with poly(ADP-ribose), so a potential interaction between HUWE1 and the important DNA repair protein poly(ADP-ribose) polymerase 1 was studied as a possible mechanism underlying the effects on DNA damage and cell survival. This study identified that HUWE1 is an important regulator of multiple DNA repair pathways, and a modulator of cell responsiveness to treatments with poly(ADP-ribose) polymerase inhibitors and DNA damaging agents. This suggests that HUWE1 is a possible target for improving ovarian cancer therapies.
Open Access
The Evolutionary History Of The Cleptoparasitic Bee Genus Nomada With An Emphasis On The Species Of Eastern North America
(2024-11-07) Odanaka, Katherine Akemi; Rehan, Sandra
Approximately 12% of all bee species found in North America are brood parasitic. Of the different genera of cleptoparasitic bees, the most diverse and taxonomically challenging is the genus Nomada, which has approximately 800 species and is found on every continent except Antarctica. Little is known regarding Nomada biology, evolution, and population dynamics and further complicating studying Nomada is inconsistency in species concepts due to variation within and between species and the lack of updated taxonomic tools needed to identify species. This dissertation aims to address gaps pertaining to the evolutionary history, biogeography, and population dynamics of Nomada at both a global and North American level. The first global phylogeny and biogeography for the genus is presented here. Nomada was found to have a Holarctic origin around 65 million years ago and subsequently dispersed into the southern hemisphere at least three times while also dispersing throughout the Nearctic and Palearctic multiple times over various land bridges. To explore phylogeographic patterns and population genetic structure and diversity, the North American endemic species Nomada articulata was used as a model, a first for a cleptoparasitic bee. It is suggested that the Wisconsinan glaciation influenced N. articulata populations by dividing the species along an east-west gradient but over time, these populations dispersed, creating a single panmictic population that persists to the present. Furthermore, N. articulata is suggested to be a species complex with two possible new and unknown species in the southern United States. Lastly, newly updated keys are provided with the aim to help remedy the lack of updated species level keys for the genus in eastern North America. As a result, fully illustrated keys are provided for five of the seven Nomada species groups found in eastern Canada and the United States covering male and female characters for 15 different species. Additionally, one new synonymy is presented and updated information pertaining to the biology, evolution, and taxonomy is included. The research presented here provides an important first step towards understanding the evolution and diversification of Nomada and how, more broadly, cleptoparasitic bees became one of the most successful groups of bees.
Open Access
An examination of facilitative interactions between shrubs and local desert organisms within the desert ecosystems of southern California
(2024-11-07) Zuliani, Mario; MacDonald, Suzanne
In recent decades, facilitative interactions have been the focus of ecological studies, specifically those with foundational shrubs within drylands. However, little has been done to analyze how the density of shrubs influence the frequency of these interactions. Here, I aim to establish shrub density as a crucial measure in ecology and facilitative interactions. I begin with a systematic review and meta-analysis (Chapter 2) to quantify the current understanding of shrub density on local animal abundance. A full text review of shrub density and animal abundance was conducted with 113 unique observations utilized in the meta-analysis. Shrub density positively predicted animal abundance, specifically within grassland and desert ecosystems. I conducted a greenhouse experiment to examine the effects of temperature on native and invasive plant species germination (Chapter 3). Temperatures were manipulated to simulate cooler microclimates and high temperature extremes. Increasing temperatures reduced germination, establishment, and biomass of all plant species, with the invasive species suffering more than natives. I conducted a telemetry experiment to determine the effects of shrub densities on an endangered species (Chapter 4). Telemetry data was combined with satellite-based counts of shrub density. Increasing shrub densities positively predicted the presence of G. sila individuals above ground. A field experiment comparing high and no shrub densities across aridity ecosystems in Southern California was conducted (Chapter 5). Camera traps were used at distinct ecosystems across southern California to determine vertebrate association. Areas of high shrub density influenced the richness and evenness of vertebrate species, while increasing temperatures negatively impacted vertebrate abundance, richness, and evenness. Finally, I tested if facilitative associations could be replicated with artificial structures (Chapter 6). Mimics were deployed within California drylands. Camera traps were utilized to estimate vertebrate communities, with temperature pendants used to contrast microclimates. Shrubs and mimics had similar microclimates, reducing the temperature underneath their canopy. Mimics displayed a higher vertebrate abundance and richness than open areas. These findings illustrate that facilitation between shrubs and dryland communities are crucial interactions. These findings suggest that shrub density is a critical measure, increasing the likelihood of interactions and possible benefits associated with shrubs.
Open Access
Potential Roles Of Circska3 In Epithelial Ovarian Cancer Development
(2024-11-07) Agbede, Oluwatobi Michael; Chun Peng
Ovarian cancer (OC) is the deadliest gynecologic cancer, of which epithelial ovarian cancer (EOC) is the most prevalent and aggressive form. Circular RNAs (circRNAs) have emerged as potential therapeutic targets and biomarkers in various cancers, including EOC. This study investigated circSKA3, which was identified to be overexpressed in high-grade serous carcinoma (HGSC). Using qRT-PCR, we confirmed upregulation in HGSC tissues compared to less aggressive subtypes. Functional assays revealed circSKA3's tumor-promoting effects in HGSC cell lines, enhancing proliferation, migration, and clonogenicity. We identified its possible involvement in epithelial-mesenchymal transition (EMT) and dysregulation of cellular metabolism, contributing to increased aggressiveness in HGSC. Protein pulldown assays and mass spectrometry analysis also identified potential interacting proteins. We then predicted circSKA3-miRNA-mRNA network to explore potential miRNA-mRNA crosstalk dysregulated by circSKA3. These findings suggest that circSKA3, via a complex regulatory interplay of proteins and miRNA interactions, modulates EMT and cellular metabolism to drive HGSC malignancy.
Open Access
Using Fecal Hormones to Determine Reproductive State and Female Choice in Wild Vervet Monkeys (Chlorocebus pygerythrus)
(2024-11-07) Rademacher, Lina Ann; Schoof, Valerie
This study aimed to assess the feasibility of using fecal hormones to identify female reproductive state in wild vervet monkeys (Chlorocebus pygerythrus) in order to gain insight on changes in female mate choice. We followed a single study group with 16 adult/subadult females at Lake Nabugabo, Uganda over a 5-month field season in 2022. During this time, 248 fecal samples and 309 hours of behavioral observations were collected and used in analyses. Mean gestation length from hormonal data was 164.33 ± 18.09 days (range 134 – 187 days) based on six females that conceived during the study. However, gaps in fecal sample collection highlight that, while feasible, greater sampling effort is needed in future studies using fecal hormones. Generally, behavioral results indicate females displayed more proceptive and receptive behaviors towards males while ovulatory and appeared to prefer high-ranking males. However, there was no effect of female dominance rank on mate choice.
Open Access
Plant-pollinator relationships of culturally significant food and medicine plants in the Great Lakes Region
(2024-11-07) Gibson, Shelby Dawn; Colla, Shelia; Fitch, Gordon
Wild plant and pollinator populations have experienced declines globally in recent years. While species declines have often been examined for their impacts ecologically and economically, little is known about how species declines will impact humans culturally. All Indigenous cultures in North America use plant species for food and medicine. Plants considered important to a specific culture are known as cultural keystone species, meaning they are significant to cultural identity. With global declines in plant and pollinator species, this study aims to determine the role of wild pollinators to cultural keystone plant species. Availability of information on breeding systems and important pollinators is crucial information for management of plant species, especially if a plant species is experiencing population declines (e.g. at-risk, endangered). A literature review was conducted to determine what is known about the breeding systems of cultural keystone species in eastern North America. The results show 50% of the culturally significant plants lack any information regarding the breeding system. A plant-pollinator network was constructed based on observations in the field on the Three Sisters (corn, beans, squash) garden. The common eastern bumble bee (Bombus impatiens) and hoary squash bee (Xenoglossa pruinosa) were important pollinators in this system. A breeding experiment including diurnal and nocturnal video recording was conducted to determine the most frequent visitors of sacred Tobacco (Nicotiana rustica), and to determine the degree to which N. rustica relies on insect pollination. Lastly, a breeding experiment was conducted on common bearberry (Arctostaphylos uva-ursi) in eastern Ontario, Canada to determine the plant's reliance on insect pollination. In conclusion, wild pollinators are important to the continued success of cultural keystone species, and therefore to the cultures that use these plants. Including multiple ways of knowing and practicing ethical ecology in the future is part of ensuring biocultural conservation.