Dynamics and functions of bacterial communities in bark, charcoal and sand filters treating greywater

This study explored the effects of greywater application on the dynamics and functions of biofilms developed in bark, activated charcoal and sand filters used for removal of organic matter and nitrogen. Duplicate columns (20 cm diameter, 60 cm deep) were packed with bark, charcoal or sand with effective size 1.4 mm and uniformity coefficient 2.2, and dosed with 32 L m(-2) day(-1) of an artificial greywater (14 g BOD5 m(-2) day(-1)) for 116 days. Potential respiration rate (PRR), determined in filter samples after addition of excess glucose, and bacterial diversity and composition, analysed by 454-pyrosequencing of bacterial 16S ribosomal DNA, were measured at different times and depths in the filters. The bark and charcoal filters were more efficient in removing BOD5 than the sand (98, 97% and 75%, respectively). The highest PRR in the 0-2 cm layer of the columns on day 84 was found in the bark filters, followed by the charcoal and sand filters (632 ± 66, 222 ± 34 and 56 ± 2 mg O2 L(-1), respectively; n = 2). Bacterial community in the bark filters showed the highest richness. The charcoal and sand filters both developed more diverse and dynamic (changing over time and depth) bacterial communities than the bark. In addition to the greywater, the lignocelluosic composition of the bark and its lower pH probably selected for the bacterial community structure and the organic content provided additional substrate, as shown by its higher PRR and its different nitrifying bacterial genera. In the oligotrophic charcoal and sand, the composition of the greywater itself defined the bacterial community. Thus, the initially low bacterial biomass in the latter filters was enriched over time, allowing a diversified bacterial community to develop. The top layers of the bark and charcoal filters displayed a high dominance of Rhizobium, Pseudomonas and Acinetobacter, which were less evident in the 60 cm layer, whereas in the sand filters these genera were prominent at both 0-2 cm and 60 cm. The PRR, bacterial diversity and composition profiles indicated that organic matter degradation occurred mainly in the top 20 cm of the bark and charcoal filters. This means that bark and charcoal filters could be designed to be shallower than sand filters.