Bacterial community responses to increasing ammonia concentrations in model recirculating vertical flow saline biofilters

This study represents the first analysis of ammonia removal and bacterial communities in gravel biofilters treating saline wastewater and is of relevance to the growing inland marine aquaculture industry. As part of a study to gain greater understanding of the microbial processes occurring in a newly constructed limestone gravel wetland at a commercial marine fish farm, this study was designed to establish the ammonia removal capacity of model biofilters treating saline aquaculture wastewater and to investigate changes to total bacterial communities and ammonia-oxidizing bacterial communities as the biofilters are exposed to increasing ammonia concentrations. Three replicate laboratory-scale gravel biofilters were constructed and the limits of nitrification capacity were tested by dosing with aquaculture wastewater supplemented with increasing amounts of ammonium chloride. The experiment was run over a 12-week period with the water temperature between 24.5 and 28 °C and salinity between 28 and 38 ppt. Greater than 97% ammonia removal in each weekly treatment period was observed with ammonia concentrations of up to 600 ppm. At higher concentrations of ammonia, a lower percentage of ammonia was removed, and on occasion nitrite accumulation was observed. A drop in the number of operational taxonomic units (OTUs) detected in the bacterial community as measured by 16s rRNA T-RFLP was observed concurrent with the decrease in percentage ammonia removal. T-RFLP of the amoA gene showed the experimental biofilters to be dominated by three different OTUs of ammonia-oxidizing bacteria. A synchronous successional pattern among these three ammonia oxidizers was observed. The three OTUs were identified as belonging to three different nitrosomonad clusters. This study demonstrates that the vertical flow gravel biofilters have the ability to treat saline aquaculture wastewater that has a high ammonia concentration and that the microbial community within saline biofilters has the capacity to adapt to changing ammonia levels while maintaining nitrification activity.