Effect of biofilm on fluorescence measurements derived from fast repetition rate fluorometers

Abstract

This study evaluates, for the first time, the influence of biofilms on single and double optical window (SOW and DOW, respectively) fast repetition rate fluorometer (FRRF) measurements of microalgal photosystem-II initial fluorescence (F0), maximum fluorescence (Fm), variable fluorescence (Fv = Fm – F0), quantum yield (Fv/Fm) and functional absorption cross section (σPSII)]. Biofilms with chlorophyll > 0.1 μg cm−2 and > 0.3 μgcm−2 on SOW and DOW, respectively, produced a substantial increase in fluorescence. However, the relative magnitude of biofouling effects depended on sample chlorophyll concentrations, being more critical at concentrations < 1 mg m−3. In DOW-FRRF, biofilms affected F0 (increased) and Fv/Fm (decreased) but not Fv and σPSII, whereas in SOW-FRRF, biofilms increased fluorescence and showed a variable effect on Fv/Fm and σPSII, because only biofilms on SOW attained actual Fm. As a result, the biofilm effect was substantial on SOW-FRRF measurements. On the other hand, the neutral-density filters (representing non-chlorophyll containing biofilms) with different transmission levels reduced the fluorescence signal. Correction procedures for the above photosystem-II parameters are proposed here.