Use of image analysis to determine algal and bacterial biomass in a high rate algal pond following Percoll fractionation.

This paper reports the results of work to allow a more accurate enumeration, and gravimetric determination, of the relative proportions of algae and bacteria within the floccular matrix present in a High Rate Algal Pond (HRAP). Methodology involving a combination of physical and chemical treatments was used and the resulting material was separated using density gradient centrifugation. The "fractions" were analysed using microscopical image analysis. Each "fraction" was categorised as containing bacteria only, algae only or an association of bacteria and algae, the relative proportions of which were then determined by image analysis to enable a "true" gravimetric determination of the algal and bacterial components of HRAP biomass for the first time. The biomass from HRAPs operated both outdoors and in a glasshouse was examined over complete operational seasons to investigate environmental effects as well as possible effects of variation in COD loading rate and retention time on cell biovolumes and cell quotients. In this study, the accurate assessment of both algal and bacterial biomass, determined as cell volume or dry matter, in addition to measurement of carbon and nitrogen has enabled the calculation of cell quotients. This allows a direct comparison between these values obtained from a nutrient-rich system and those published values obtained from systems with various nutrient status. Conversion factors obtained may also be of value for deriving inputs for computer models for the design and operation of high rate algal ponds.