Brittany G. Perrotta

Per- and polyfluoroalkyl substances (PFAS) are widespread contaminants of global concern due to their persistence, biomagnification potential in aquatic environments, and mounting evidence of toxic effects. Emerging experimental evidence indicates that chronic exposure to PFAS can alter the composition and function of a host organism’s microbiome. To test this in the field, we analyzed microbiomes of larval and adult aquatic insects, along with riparian spiders, from a stream receiving industrial wastewater with elevated concentrations of PFAS from a food processing facility. Spiders consume substantial aquatic insect biomass globally and are frequently used as biosentinels for tracking aquatic-derived contamination in riparian ecosystems. Here, we found that larval and adult insects and spiders had 5-, 1.4- and 5-fold higher concentrations, respectively, of PFAS downstream than upstream of the outfall. Downstream spider microbiomes had unique community structure, higher diversity, and increased predicted representation of xenobiotic degradation and metabolism gene functions compared to paired spider microbiomes upstream of the wastewater outfall. Given the abundance and distribution of both PFAS and spiders globally, our findings provide insight into how host microbiomes may respond to ubiquitous, chronic wastewater mixtures with elevated PFAS and provide insight into evaluating PFAS exposure across aquatic-riparian ecosystems.

Larval aquatic insects harbour diverse microbiomes critical for physiology and development, yet the effects of metamorphosis combined with environmental exposures on this community remain unknown. Herein we examined bacterial microbiome composition and predicted function across larval and adult stages of three families of aquatic insects and riparian spider predators, collected upstream and downstream of wastewater treatment plants (WWTPs) in Ontario, Canada. Insect host developmental stage was the primary driver of microbiome structure. Larval microbiomes were more diverse and showed higher abundances in pathways reflecting active feeding. Larval and adult populations of holometabolous insects shared few ASVs (3.8%–12.6% of the combined ASV pool), whereas a larger proportion was shared in hemimetabolous insects (23.4%). WWTP effluent exposures led to selective microbial filtering rather than broad community restructuring. Macroinvertebrate and spider microbiomes sampled downstream were characterized by higher abundances in bacterial taxa associated with wastewaters and increased xenobiotic degradation pathways. Such impacts on spider microbiomes provide evidence that ecological impacts of effluents cross the aquatic‐terrestrial boundary. Lastly, a significant decrease of vertically transmitted endosymbionts ( Rickettsiella , Wolbachia ) was detected in aquatic insects, representing a pathway through which effluent pollution may affect host fitness without detectably altering overall microbiome diversity.

We evaluated the bioaccumulation and transfer of per- and polyfluoroalkyl substances (PFAS) in a stream food web contaminated by a food processing facility. Abiotic (i.e., water, sediment, and foam) and biotic (i.e., algae, aquatic insect larvae and adults, fish, and riparian spiders) matrices were sampled upstream and downstream of the facility's wastewater outfall. Compared with upstream, PFAS concentrations were 600-fold higher in downstream water (mean ∑₄₀PFAS 3.67 ng mL^-1 ± 0.48 (standard error)) and reflected inputs from the outfall, with 6:2 fluorotelomer sulfonate (6:2 FTS) dominating the PFAS profile. Within the aquatic food web, perfluorooctanesulfonate (PFOS) was the most biomagnified, and 6:2 FTS was the most biodiluted. In contrast, insect-mediated transfer of PFAS to riparian spiders showed trophic enrichment of 6:2 FTS and dilution of PFOS. We observed significant positive associations between phospholipid membrane-water partition coefficient (log K_MW) and perfluoroalkyl carboxylate (PFCA) chain length on bioaccumulation across most biological matrices, demonstrating that these chemical parameters are predictive of PFAS bioaccumulation potential in the field. Our research reveals important differences in aquatic versus terrestrial exposure for certain PFAS and that biological processes (e.g., trophic interactions and metamorphosis) and chemical properties (e.g., chain length, log K_MW, and concentration) control PFAS uptake, bioaccumulation, and transfer in linked freshwater and terrestrial ecosystems.

Defoliation by eastern spruce budworm is one of the most important natural disturbances in Canadian boreal and hemi‐boreal forests with annual area affected surpassing that of fire and harvest combined, and its impacts are projected to increase in frequency, severity, and range under future climate scenarios. Deciding on an active management strategy to control outbreaks and minimize broader economic, ecological, and social impacts is becoming increasingly important. These strategies differ in the degree to which defoliation is suppressed, but little is known about the downstream consequences of defoliation and, thus, the implications of management. Given the disproportionate role of headwater streams and their microbiomes on net riverine productivity across forested landscapes, we investigated the effects of defoliation by spruce budworm on headwater stream habitat and microbiome structure and function to inform management decisions. We experimentally manipulated a gradient of defoliation among 12 watersheds during a spruce budworm outbreak in the Gaspésie Peninsula, Québec, Canada. From May through October of 2019–2021, stream habitat (flow rates, dissolved organic matter [DOM], water chemistry, and nutrients), algal biomass, and water temperatures were assessed. Bacterial and fungal biofilm communities were examined by incubating six leaf packs for five weeks (mid‐August to late September) in one stream reach per watershed. Microbiome community structure was determined using metabarcoding of 16S and ITS rRNA genes, and community functions were examined using extracellular enzyme assays, leaf litter decomposition rates, and taxonomic functional assignments. We found that cumulative defoliation was correlated with increased streamflow rates and temperatures, and more aromatic DOM (measured as specific ultraviolet absorbance at 254 nm), but was not correlated to nutrient concentrations. Cumulative defoliation was also associated with altered microbial community composition, an increase in carbohydrate biosynthesis, and a reduction in aromatic compound degradation, suggesting that microbes are shifting to the preferential use of simple carbohydrates rather than more complex aromatic compounds. These results demonstrate that high levels of defoliation can affect headwater stream microbiomes to the point of altering stream ecosystem productivity and carbon cycling potential, highlighting the importance of incorporating broader ecological processes into spruce budworm management decisions.

Freshwater ecosystems are exposed to engineered nanoparticles (NPs) through discharge from wastewater and agricultural runoff. We conducted a 9-month mesocosm experiment to examine the combined effects of chronic NP additions on insect emergence and insect-mediated contaminant flux to riparian spiders. Two NPs (copper, gold, plus controls) were crossed by two levels of nutrients in 18 outdoor mesocosms open to natural insect and spider colonization. We collected adult insects and two riparian spider genera, Tetragnatha and Dolomedes, for 1 week on a monthly basis. We estimated a significant decrease in cumulative insect emergence of 19% and 24% after exposure to copper and gold NPs, irrespective of nutrient level. NP treatments led to elevated copper and gold tissue concentrations in adult insects, which resulted in terrestrial fluxes of metals. These metal fluxes were associated with increased gold and copper tissue concentrations for both spider genera. We also observed about 25% fewer spiders in the NP mesocosms, likely due to reduced insect emergence and/or NP toxicity. These results demonstrate the transfer of NPs from aquatic to terrestrial ecosystems via emergence of aquatic insects and predation by riparian spiders, as well as significant reductions in insect and spider abundance in response to NP additions.

Freshwater ecosystems are exposed to engineered nanoparticles through municipal and industrial wastewater-effluent discharges and agricultural non-point source runoff. Because previous work has shown that engineered nanoparticles from these sources can accumulate in freshwater algal assemblages, we hypothesized that nanoparticles may affect the biology of primary consumers by altering the processing of two critical nutrients associated with growth and survivorship, nitrogen and phosphorus. We tested this hypothesis by measuring the excretion rates of nitrogen and phosphorus of Physella acuta, a ubiquitous pulmonate snail that grazes heavily on periphyton, exposed to either copper or gold engineered nanoparticles for six months in an outdoor wetland mesocosm experiment. Chronic nanoparticle exposure doubled nutrient excretion when compared to the control. Gold nanoparticles increased nitrogen and phosphorus excretion rates more than copper nanoparticles but overall, both nanoparticles led to higher consumer excretion, despite contrasting particle stability and physio-chemical properties. Snails in mesocosms enriched with nitrogen and phosphorus had overall higher excretion rates than ones in ambient (no nutrients added) mesocosms. Stimulation patterns were different between nitrogen and phosphorus excretion, which could have implications for the resulting nutrient ratio in the water column. These results suggest that low concentrations of engineered nanoparticles could alter the metabolism of consumers and increase consumer-mediated nutrient recycling rates, potentially intensifying eutrophication in aquatic systems, e.g. the increased persistence of algal blooms as observed in our mesocosm experiment.

Obesity is a risk factor for colorectal cancer (CRC), accounting for more than 14% of CRC incidence. Microbial dysbiosis and chronic inflammation are common characteristics in both obesity and CRC. Human and murine studies, together, demonstrate the significant impact of the microbiome on governing energy metabolism and CRC development; yet, little is understood about the contribution of the microbiome to development of obesity-associated CRC as compared to non-obese individuals. In this study, we conducted a meta-analysis using five publicly available stool and tissue-based 16S rRNA and whole genome sequencing (WGS) data sets of CRC microbiome studies. High-resolution analysis was employed for 16S rRNA data using Resphera Insight, which allowed us to achieve species-level information to compare with WGS. Characterization of the confounders between studies, 16S rRNA variable region, and sequencing method, did not reveal any significant effect on alpha diversity in CRC prediction. Both 16S rRNA and WGS were equally variable in their ability to predict CRC. Results from community structure and composition analysis confirmed lower diversity in obese individuals without CRC; however, no universal differences were found in diversity between obese and non-obese individuals with CRC. When examining taxonomic differences, the probability of being classified as CRC did not change significantly in obese individuals for all taxa tested. However, random forest classification was able to distinguish CRC and non-CRC stool when body mass index was added to the model. Overall, microbial dysbiosis was not a significant factor in explaining the higher risk of colon cancer among individuals with obesity.