Arctic microbial and next-generation sequencing approach for bacteria in snow and frost flowers: selected identification, abundance and freezing nucleation

During the spring of 2009, as part of the Ocean-Atmosphere-Sea Ice-Snowpack (OASIS) campaign in Barrow, Alaska, USA, we examined the identity, population diversity, freezing nucleation ability of the microbial communities of five different snow types and frost flowers. In addition to the culturing and gene-sequence-based identification approach, we utilized a state-of-the-art genomic next-generation sequencing (NGS) technique to examine the diversity of bacterial communities in Arctic samples. Known phyla or candidate divisions were detected (11-18) with the majority of sequences (12.3-83.1 %) belonging to one of the five major phyla: Proteobacteria, Actinobacteria, Bacteroidetes, Firmicutes, and Cyanobacteria. The number of genera detected ranged from, 101-245. The highest number of cultivable bacteria was observed in frost flowers (FFs) and accumulated snow (AS) with 325 +/- 35 and 314 +/- 142 CFUmL(-1), respectively; and for cultivable fungi 5 +/- 1 CFUmL(-1) in windpack (WP) and blowing snow (BS). Morphology/elemental composition and ice-nucleating abilities of the identified taxa were obtained using high resolution electron microscopy with energy-dispersive X-ray spectroscopy and ice nucleation cold-plate, respectively. Freezing point temperatures for bacterial isolates ranged from -20.3 +/- 1.5 to -15.7 +/- 5.6 degrees C, and for melted snow samples from -9.5 +/- 1.0 to -18.4 +/- 0.1 degrees C. An isolate belonging to the genus Bacillus (96% similarity) had ice nucleation activity of -6.8 +/- 0.2 degrees C. Comparison with Montreal urban snow, revealed that a seemingly diverse community of bacteria exists in the Arctic with some taxa possibly originating from distinct ecological environments. We discuss the potential impact of snow microorganisms in the freezing and melting process of the snowpack in the Arctic.