Exploring the distance between nitrogen and phosphorus limitation in mesotrophic surface waters using a sensitive bioassay

Abstract. The balance in microbial net consumption of nitrogen and phosphorus was investigated in samples collected in two mesotrophic coastal environments: the Baltic Sea (Tvärminne field station) and the North Sea (Espegrend field station). For this, we have refined a bioassay based on the response in alkaline phosphatase activity (APA) over a matrix of combinations in nitrogen and phosphorus additions. This assay not only provides information on which element (N or P) is the primary limiting nutrient, but also gives a quantitative estimate for the excess of the secondary limiting element (P⁺ or N⁺, respectively), as well as the ratio of balanced net consumption of added N and P over short timescales (days). As expected for a Baltic Sea late spring–early summer situation, the Tvärminne assays (n =  5) indicated N limitation with an average P⁺ =  0.30 ± 0.10 µM-P, when incubated for 4 days. For short incubations (1–2 days), the Espegrend assays indicated P limitation, but the shape of the response surface changed with incubation time, resulting in a drift in parameter estimates toward N limitation. Extrapolating back to zero incubation time gave P limitation with N⁺ ≈  0.9 µM-N. The N : P ratio (molar) of nutrient net consumption varied considerably between investigated locations: from 2.3 ± 0.4 in the Tvärminne samples to 13 ± 5 and 32 ± 3 in two samples from Espegrend. Our assays included samples from mesocosm acidification experiments, but statistically significant effects of ocean acidification were not found by this method.