Impacts of sea ice melting procedures on measurements of microbial community structure

Abstract

Microorganisms play critical roles in sea ice biogeochemical processes. However, microbes living within sea ice can be challenging to sample for scientific study. Because most techniques for microbial analysis are optimized for liquid samples, sea ice samples are typically melted first, often applying a buffering method to mitigate osmotic lysis. Here, we tested commonly used melting procedures on three different ice horizons of springtime, first year, land-fast Arctic sea ice to investigate potential methodological impacts on resulting measurements of cell abundance, photophysiology, and microbial community structure as determined by 16S and 18S rRNA gene amplicon sequencing. Specifically, we compared two buffering methods using NaCl solutions (“seawater,” melting the ice in an equal volume of 35-ppt solution, and “isohaline,” melting with a small volume of 250-ppt solution calculated to yield meltwater at estimated in situ brine salinity) to direct ice melting (no buffer addition) on both mechanically “shaved” and “non-shaved” samples. Shaving the ice shortened the melting process, with no significant impacts on the resulting measurements. The seawater buffer was best at minimizing cell lysis for this ice type, retaining the highest number of cells and chlorophyll a concentration. Comparative measurements of bacterial (16S) community structure highlighted ecologically relevant subsets of the community that were significantly more abundant in the buffered samples. The results for eukaryotic (18S) community structure were less conclusive. Taken together, our results suggest that an equivalent-volume seawater-salinity buffered melt is best at minimizing cell loss due to osmotic stress for springtime Arctic sea ice, but that either buffer will reduce bias in community composition when compared to direct melting. Overall, these findings indicate potential methodological biases that should be considered before developing a sea ice melting protocol for microbiological studies and afterwards, when interpreting biogeochemical or ecological meaning of the results.