Underestimation of microzooplankton grazing in dilution experiments due to inhibition of phytoplankton growth

Microzooplankton dilution grazing experiments conducted in phytoplankton rich waters, particularly in polar and subpolar seas, often result in calculation of nonsignificant or negative grazing coefficients. We hypothesized that preparation of filtered seawater (FSW) from water containing high biomass of phytoplankton results in release of allelochemicals that inhibit phytoplankton growth, lowering the net growth of phytoplankton in the more diluted treatments. We tested this hypothesis during blooms of Skeletonema marinoi and Phaeocystis pouchetii in a nutrient-enriched mesocosm in the Raunefjord, Norway. During the S. marinoi bloom, inhibition of phytoplankton growth occurred in the diluted treatments. Simultaneously the concentration of total as well as dissolved polyunsaturated aldehydes (PUAs) was elevated. Passage of the FSW through a carbon cellulose cartridge to remove dissolved organic material reduced, or eliminated, the inhibition. In the early phase of the P. pouchetii bloom that followed the diatom bloom in the mesocosm, PUA concentration was relatively low and the untreated FSW had a less drastic, but often significant, inhibitory effect on phytoplankton growth. Laboratory experiments with cultures of S. marinoi and P. pouchetii confirmed that material present in filtrate prepared from diluted cultures was self-inhibitory. Many phytoplankters, particularly during late stages of a bloom, produce inhibitory metabolites that may be released during filtration of the relatively large volumes of seawater needed for dilution experiments. Under some conditions, dilution grazing experiments may underestimate phytoplankton growth coefficients and microzooplankton grazing coefficients.