Size really does matter: effects of filter fractionation on microbial community structure in a model oxygen minimum zone.

The Scientific Committee on Oceanographic Research (SCOR) Working Group 144 “Microbial Community Responses to Ocean Deoxygenation” workshop held in Vancouver, British Columbia in July 2014 had the primary objective of kick-starting the establishment of a minimal core of technologies, techniques and standard operating procedures (SOPs) to enable compatible process rate and multi-molecular data (DNA, RNA and protein) collection in marine oxygen minimum zones (OMZs) and other oxygen starved waters. Experimental activities conducted in Saanich Inlet, a seasonally anoxic fjord on Vancouver Island British Columbia, were designed to compare and cross-calibrate in situ sampling devices (McLane PPS system) with conventional bottle sampling and incubation methods. Bottle effects on microbial community composition, and activity were tested using different filter combinations and sample volumes to compare PPS/IPS (0.4 µm) versus Sterivex (0.22 µm) filtration methods with and without prefilters (2.7 µm). Resulting biomass was processed for small subunit ribosomal RNA gene sequencing across all three domains of life on the 454 platform followed by downstream community structure analyses. Significant community shifts occurred within and between filter fractions for in situ versus on-ship processed samples. For instance, the relative abundance of several bacterial phyla including Bacteroidetes, Delta and Gammaproteobacteria decreased five-fold on-ship when compared to in situ filtration. Similarly, experimental mesocosms showed similar community structure and activity to in situ filtered samples indicating the need to cross-calibrate incubations to constrain bottle effects. In addition, alpha and beta diversity significantly changed as function of filter size and volume, as well as the operational taxonomic units identified using indicator species analysis for each filter size. Our results provide statistical support that microbial community structure is systematically biased by filter fraction methods and highlight the need for establishing compatible techniques among researchers that facilitate comparative and reproducible science for the whole community.