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Bioindicator “fingerprints” of methane-emitting thermokarst features in Alaskan soils

Frontiers in Microbiology

Schambach, Jenna Y.; Bennett, Haley L.; Bays, Nathan R.; Yang, Jihoon; Kolker, Stephanie D.; Ricken, Bryce; Sammon, Jason P.; Mascarenas, Monica R.; Whiting, Joshua J.; Eberling, Naomi A.; Mays, Wittney D.; Anthony, Katey W.; Miller, Philip R.; Smallwood, Chuck R.

Permafrost thaw increases the bioavailability of ancient organic matter, facilitating microbial metabolism of volatile organic compounds (VOCs), carbon dioxide, and methane (CH4). The formation of thermokarst (thaw) lakes in icy, organic-rich Yedoma permafrost leads to high CH4 emissions, and subsurface microbes that have the potential to be biogeochemical drivers of organic carbon turnover in these systems. However, to better characterize and quantify rates of permafrost changes, methods that further clarify the relationship between subsurface biogeochemical processes and microbial dynamics are needed. In this study, we investigated four sites (two well-drained thermokarst mounds, a drained thermokarst lake, and the terrestrial margin of a recently formed thermokarst lake) to determine whether biogenic VOCs (1) can be effectively collected during winter, and (2) whether winter sampling provides more biologically significant VOCs correlated with subsurface microbial metabolic potential. During the cold season (March 2023), we drilled boreholes at the four sites and collected cores to simultaneously characterize microbial populations and captured VOCs. VOC analysis of these sites revealed “fingerprints” that were distinct and unique to each site. Total VOCs from the boreholes included > 400 unique VOC features, including > 40 potentially biogenic VOCs related to microbial metabolism. Subsurface microbial community composition was distinct across sites; for example, methanogenic archaea were far more abundant at the thermokarst site characterized by high annual CH4 emissions. The results obtained from this method strongly suggest that ∼10% of VOCs are potentially biogenic, and that biogenic VOCs can be mapped to subsurface microbial metabolisms. By better revealing the relationship between subsurface biogeochemical processes and microbial dynamics, this work advances our ability to monitor and predict subsurface carbon turnover in Arctic soils.

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Biotic countermeasures that rescue Nannochloropsis gaditana from a Bacillus safensis infection

Frontiers in Microbiology

Humphrey, Brittany M.; Mackenzie, Morgan; Lobitz, Mia; Schambach, Jenna Y.; Lasley, Greyson; Kolker, Stephanie D.; Ricken, Bryce; Bennett, Haley L.; Williams, Kelly P.; Smallwood, Chuck R.; Cahill, Jesse L.

The natural assemblage of a symbiotic bacterial microbiome (bacteriome) with microalgae in marine ecosystems is now being investigated as a means to increase algal productivity for industry. When algae are grown in open pond settings, biological contamination causes an estimated 30% loss of the algal crop. Therefore, new crop protection strategies that do not disrupt the native algal bacteriome are needed to produce reliable, high-yield algal biomass. Bacteriophages offer an unexplored solution to treat bacterial pathogenicity in algal cultures because they can eliminate a single species without affecting the bacteriome. To address this, we identified a highly virulent pathogen of the microalga Nannochloropsis gaditana, the bacterium Bacillus safensis, and demonstrated rescue of the microalgae from the pathogen using phage. 16S rRNA amplicon sequencing showed that phage treatment did not alter the composition of the bacteriome. It is widely suspected that the algal bacteriome could play a protective role against bacterial pathogens. To test this, we compared the susceptibility of a bacteriome-attenuated N. gaditana culture challenged with B. safensis to a N. gaditana culture carrying a growth-promoting bacteriome. We showed that the loss of the bacteriome increased the susceptibility of N. gaditana to the pathogen. Transplanting the microalgal bacteriome to the bacteriome-attenuated culture reconstituted the protective effect of the bacteriome. Finally, the success of phage treatment was dependent on the presence of beneficial bacteriome. This study introduces two synergistic countermeasures against bacterial pathogenicity in algal cultures and a tractable model for studying interactions between microalgae, phages, pathogens, and the algae microbiome.

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Electrochemical aptamer-based sensors: leveraging the sensing platform for minimally-invasive microneedle measurements and fundamental exploration of sensor biofouling dynamics

Downs, Alexandra M.; Miller, Philip R.; Bolotsky, Adam; Staats, Amelia M.; Weaver, Bryan M.; Bennett, Haley L.; Tiwari, Sidhant; Kolker, Stephanie D.; Wolff, Nathan P.; Polsky, Ronen; Larson, Steven R.; Coombes, Kenneth R.; Sawyer, Patricia S.

The ability to track the concentrations of specific molecules in the body in real time would significantly improve our ability to study, monitor, and respond to diseases. To achieve this, we require sensors that can withstand the complex environment inside the body. Electrochemical aptamer-based sensors are particularly promising for in vivo sensing, as they are among the only generalizable sensing technologies that can achieve real-time molecular monitoring directly in blood and the living body. In this project, we first focused on extending the application space of aptamer sensors to support minimally-invasive wearable measurements. To achieve this, we developed individually-addressable sensors with commercial off-the-shelf microneedles. We demonstrated sensor function in buffer, blood, and porcine skin (a common proxy for human skin). In addition to the applied sensing project, we also worked to improve fundamental understanding of the aptamer sensing platform and how it responds to biomolecular interferents. Specifically, we explored the interfacial dynamics of biofouling – a process impacting sensors placed in complex fluids, such as blood.

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