IMPACTS OF PLASTIC POLLUTION ON A PELAGIC MARINE MAMMAL, THE NORTHERN ELEPHANT SEAL

As plastic pollution increases, top marine predators such as marine mammals are becoming increasingly susceptible to plastic particles and their additives. Plastic particles have been found in gastrointestinal tracts and scat of many marine mammals, and quantifying plastic pollution in those that are pelagic can provide insight into plastic pollution in mesopelagic ecosystems that are just beginning to be analyzed. Adapting well-developed laboratory techniques for microplastic (MP) isolation (i.e. density separation and chemical digestion), I isolated MPs from the scat of the deepest diving pinniped, the northern elephant seal (NES), and found that100% of scat samples (n=11) contained high counts of MPs compared to other pinnipeds. Further, as plastic particles move through the digestive tract and potentially translocate into the circulation, body cells may be exposed to these plastic particles. Nanoplastics (NPs;\u3c1μm) have been shown to affect cell viability and redox homeostasis in fish and human cells, while the lipophilic additives bisphenol-a (BPA) and bisphenol-s (BPS) used in plastics production are known endocrine disruptors. However, the effects of plastics (NPs) and plastics additives (BPA and BPS) have not been well-studied in marine mammals. To assess the direct impacts of NPs on marine mammal cells, I exposed fibroblasts isolated from NES skin (n=6 experiments) to polystyrene NPs of two different sizes (0.05μm and 0.5μm) and concentrations (3.64x109 and 3.64x1010 particles/mL) and did not find consistent effects on morphology and viability. Cell viability, which was quantified by an MTT assay, decreased as a result of NP exposure in two experimental replicates, but these results were not reproducible. I found that NPs did not have consistent effects on the morphology or viability of NES fibroblasts, regardless of their size or concentration. Lastly, I examined the effects of plastic additives on the physiology of marine mammal blubber tissue, the primary energy depot and reservoir of lipophilic pollutants. Precision-cut NES blubber slices were exposed to BPA and BPS, alone and in combination with the lipolytic hormone epinephrine, and I assessed their effects on the blubber transcriptome. I found that while BPA and BPS treatments alone did not have a pronounced effect on gene expression, they altered the expression of several genes associated with lipid homeostasis and adipogenesis. These data suggest that NES likely ingest MPs and maybe physiologically affected by exposure to plastic particles and their associated contaminants

IMPACTS OF PLASTIC POLLUTION ON A PELAGIC MARINE MAMMAL, THE NORTHERN ELEPHANT SEAL

As plastic pollution increases, top marine predators such as marine mammals are becoming increasingly susceptible to plastic particles and their additives. Plastic particles have been found in gastrointestinal tracts and scat of many marine mammals, and quantifying plastic pollution in those that are pelagic can provide insight into plastic pollution in mesopelagic ecosystems that are just beginning to be analyzed. Adapting well-developed laboratory techniques for microplastic (MP) isolation (i.e. density separation and chemical digestion), I isolated MPs from the scat of the deepest diving pinniped, the northern elephant seal (NES), and found that100% of scat samples (n=11) contained high counts of MPs compared to other pinnipeds. Further, as plastic particles move through the digestive tract and potentially translocate into the circulation, body cells may be exposed to these plastic particles. Nanoplastics (NPs;\u3c1μm) have been shown to affect cell viability and redox homeostasis in fish and human cells, while the lipophilic additives bisphenol-a (BPA) and bisphenol-s (BPS) used in plastics production are known endocrine disruptors. However, the effects of plastics (NPs) and plastics additives (BPA and BPS) have not been well-studied in marine mammals. To assess the direct impacts of NPs on marine mammal cells, I exposed fibroblasts isolated from NES skin (n=6 experiments) to polystyrene NPs of two different sizes (0.05μm and 0.5μm) and concentrations (3.64x109 and 3.64x1010 particles/mL) and did not find consistent effects on morphology and viability. Cell viability, which was quantified by an MTT assay, decreased as a result of NP exposure in two experimental replicates, but these results were not reproducible. I found that NPs did not have consistent effects on the morphology or viability of NES fibroblasts, regardless of their size or concentration. Lastly, I examined the effects of plastic additives on the physiology of marine mammal blubber tissue, the primary energy depot and reservoir of lipophilic pollutants. Precision-cut NES blubber slices were exposed to BPA and BPS, alone and in combination with the lipolytic hormone epinephrine, and I assessed their effects on the blubber transcriptome. I found that while BPA and BPS treatments alone did not have a pronounced effect on gene expression, they altered the expression of several genes associated with lipid homeostasis and adipogenesis. These data suggest that NES likely ingest MPs and maybe physiologically affected by exposure to plastic particles and their associated contaminants

Effects of cortisol, epinephrine, and bisphenol contaminants on the transcriptional landscape of marine mammal blubber

Top ocean predators such as marine mammals are threatened by intensifying anthropogenic activity, and understanding the combined effects of multiple stressors on their physiology is critical for conservation efforts. We investigated potential interactions between stress hormones and bisphenol contaminants in a model marine mammal, the northern elephant seal (NES). We exposed precision-cut adipose tissue slices (PCATS) from blubber of weaned NES pups to cortisol (CORT), epinephrine (EPI), bisphenol A (BPA), bisphenol S (BPS), or their combinations (CORT-EPI, BPA-EPI, and BPS-EPI) ex vivo and identified hundreds of genes that were differentially regulated in response to these treatments. CORT altered expression of genes associated with lipolysis and adipogenesis, whereas EPI and CORT-EPI-regulated genes were associated with responses to hormones, lipid and protein turnover, immune function, and transcriptional and epigenetic regulation of gene expression, suggesting that EPI has wide-ranging and prolonged impacts on the transcriptional landscape and function of blubber. Bisphenol treatments alone had a weak impact on gene expression compared with stress hormones. However, the combination of EPI with bisphenols altered expression of genes associated with inflammation, cell stress, DNA damage, regulation of nuclear hormone receptor activity, cell cycle, mitochondrial function, primary ciliogenesis, and lipid metabolism in blubber. Our results suggest that CORT, EPI, bisphenols, and their combinations impact cellular, immune, and metabolic homeostasis in marine mammal blubber, which may affect the ability of marine mammals to sustain prolonged fasting during reproduction and migration, renew tissues, and mount appropriate responses to immune challenges and additional stressors