Microorganisms are the first responder to oil spills and their response provides insight into the ecological effects of oils on aquatic ecosystems. Limited information is available about the impact of oil spills on freshwater ecosystems under seasonal river-ice regimes. This study aimed to investigate the microbial response of North Saskatchewan River water to diluted bitumen (DB) and conventional crude (CC) during the freeze-thaw-refreeze cycle. In two separate experiments, equivalent to 2 L of fresh DB and CC were spilled on the ice-covered river water within a mesoscale spill tank. The microbial response (changes in abundance and diversity) to oils under the freeze, thaw, and refreeze cycles were assessed for 10 days using 16S rRNA gene sequencing. The results showed that microbial communities exhibited different responses to the DB and CC oils. The effect of oils was more pronounced than that of the freeze or thaw cycles. The river microbial community rapidly responded to both spills, which coincided with a steady increase in the organic content of water throughout the freeze-thaw-refreeze cycle. Microbial diversity increased after the DB spill, but remain unchanged after the CC spill, regardless of the cycles. A higher number of new taxa emerged during the ice-covered period, while more microbial enrichment (increase in abundance) was observed during the thaw cycle. Flavobacterium (37 ± 5%) and Pseudomonas (36 ± 4%) remained the most predominant genera post-DB and CC spill, respectively. The results of this study suggest that ice coverage of 5 cm did not prevent the microbial communities from the effects of oils. Thus, a quick clean-up response to an oil spill on ice-covered water is equally critical to avoid the effects of oils on the underlying freshwater ecosystems.
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