Morphology and LPS content for the estimation of marine bacterioplankton biomass in the Ionian Sea

The abundance, morphotypes and biomass of the bacterial assemblages were investigated in the Ionian Sea by using two different methods: the epifluorescent microscopy technique for enumerating and sizing bacterial cells, and the determination of bacterial lipopolysaccharides (LPS). Five bacterial morphotypes were distinguished: cocci, rods, coccobacilli, vibrios and spirillae. The proportions of cocci were higher than those of other morphotypes at every depth, ranging from 39% to 73%. Both rod-shaped bacteria and coccobacilli were homogenously distributed in the water column, while the proportions of vibrios were rather variable. Spirillae occurred only in surface samples and disappeared below 100 m. The two methodologies were compared: LPS concentrations showed a highly significant correlation with the bacterial numbers (P< 0.01; n= 88; r= 0.68), but not with biovolumes, and different ratios between LPS concentrations and bacterial volumes were recorded for the photic and aphotic zones (3.11 ± 1.35 and 0.96 ± 0.37 ng LPS per µm3 respectively). LPS-derived cell carbon content on average was 23 fg C cell-1, similar to the C amount derived by mean cell biovolume (19 fg C cell-1) and the biomass from two highly correlated methods (P< 0.01; n= 95; r= 0.59). Our results confirm that the widely used factor of 20 fg C cell-1 (Lee and Furhman, 1987) should be plausible for studying the biomass of the natural microbial populations in the study area. Nevertheless, the wide variability of the cell size classes, also along the whole water columns, questions the applicability of a constant conversion factor for all the marine ecosystems. Consequently, locally derived biomass estimates of bacteria are essential in order to obtain an accurate evaluation of the bacterial role in biogeochemical cycles.