Quantification of endospores in ancient permafrost using time-resolved terbium luminescence.

We describe herein a simple procedure for quantifying endospore abundances in ancient and organic-rich permafrost. We repeatedly (10x) extracted and fractionated permafrost using a tandem filter assembly composed of 3 and 0.2 µm filters. Then, the 0.2 µm filter was washed (7x), autoclaved, and the contents eluted, including dipicolinic acid (DPA). Time-resolved luminescence using Tb(EDTA) yielded a LOD of 1.46 nM DPA (6.55 × 103 endospores/mL). In review, DPA/endospore abundances were ~2.2-fold greater in older 33 ky permafrost (258 ± 36 pmol DPA gdw-1; 1.15 × 106 ± 0.16 × 106 spores gdw-1) versus younger 19 ky permafrost (p = 0.007297). This suggests that dormancy increases with permafrost age.