Estimation of nitrifying and heterotrophic bacterial activity in biofilm formed on RAS biofilter carriers by respirometry

Biofilm, the aggregates of microbial layers that are attached on the surface of biofilter carriers, plays a central role in the removal and conversion of nutrients in recirculating aquaculture systems (RAS). The bacterial activity in biofilm involves autotrophic and heterotrophic processes. These biological processes are crucial for the water quality in RAS but are difficult to monitor. In this study, we demonstrate a new method based on intermittent respirometry to selectively estimate bacterial activity in biofilm following a spike in substrate concentration. The method was tested with three different biofilter carriers (extruded polypropylene, injection molded poly-propylene and polymeric foam) from moving bed biofilm reactors in a common freshwater RAS. Oxygen consumption rates of biofilm-associated bacteria were measured in closed metabolic chambers under standard conditions. The protocol included sequential flushes of either pure tap water, or tap water spiked with nitrite, ammonium or acetate. The results showed consistent carrier-specific metabolic activities of endogenous respiration, nitrite-oxidizing bacteria (NOB), ammonia-oxidizing bacteria (AOB) and heterotrophic bacteria (HB) in biofilm. The highest activities normalized to volumetric oxygen consumption rates, were found in polymeric foam with values of 677 +/- 125, 764 +/- 156 and 166 +/- 36 g O-2.m(-3).d(-1 )for NOB, AOB and HB, respectively. The biofilter performance evaluation based on respirometric data showed satisfactory accordance with the values from substrate degradation batch kinetic tests for all three tested carriers, confirming the feasibility and robustness of the method as a means to assessing biofilter performance. Our study provides an on-site tool in biofilter performance tests and leads to a better understanding of dynamic and relationship between biofiltration and water quality.

Automated summary

Biofilm plays a crucial role in the removal and conversion of nutrients in recirculating aquaculture systems (RAS), but monitoring the bacterial activity in biofilm is difficult. In this study, a new method based on intermittent respirometry was demonstrated to selectively estimate bacterial activity in biofilm, and its feasibility and robustness were confirmed through experiments.