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A dialysis bags experiment, involving the use of natural water from eutrophic LakeMiko8ajskie (Mazurian Lake District, Poland), was performed to assess the impact of grazing pressureby heterotrophic nanoflagellates (HNF; <10 µm) on bacterial abundance, their size distribution, tax-onomic composition and activity measured by bacterial secondary production (BP) and the contribu-tion of active bacteria with an intact membrane (MEM+). Three different techniques—size-fraction-ation (<1.0 µm pore size), dilution (10×) and selective eukaryotic inhibition (cycloheximide andcolchicine)—were applied to control HNF grazing. There were significant differences in the effec-tiveness of grazing pressure limitation between the different techniques employed. The most distinctchanges in the bacterial community were found in the control and the 10× diluted treatments; in both,a considerable increase in HNF abundance was noted. In these 2 treatments, a significant decreasein both bacterial numbers and contribution of MEM+ cells, as well as an increase in the proportion ofbacterial filaments (up to 50 µm long), was observed. In contrast, the use of cycloheximide andcolchicine in combination resulted in the lowest and an almost constant HNF abundance. In addition,high numbers of bacteria with a large proportion of MEM+ cells (30 to 40%) and exponential increaseof bacterial secondary production were found. These results suggest that eukaryotic inhibition is themost useful method of limiting HNF grazing pressure and involves minimal manipulation of the watersample. Close relationships between bacteria and HNF confirm the crucial role of nanoflagellates inshaping bacterial abundance, morphology and activity.KEY WORDS: Bacteria · HNF · Grazing pressure · Dialysis bags · Filtration · Dilution · Eukaryotic inhibition

Relationships between bacteria and heterotrophic nanoflagellates in lake water examined by different techniques controlling grazing pressure