Response to Reviewer 2

Abstract

The treatment of organic wastes remains one of the key sustainability challenges facing the growing global aquaculture industry. Bioremediation systems based on coupled bioturbation-microbial processing offer a promising route for waste management. We present, for the first time, a combined biogeochemical-molecular analysis of the short-term performance of one such system that is designed to receive nitrogen-rich particulate aquaculture wastes. Using sea cucumbers (Holothuria scabra) as a model bioturbator we provide evidence that adjusting the waste C : N from 5 : 1 to 20 : 1 promoted a shift in nitrogen cycling pathways towards the dissimilatory nitrate reduction to ammonium (DNRA), resulting in net NH + 4 efflux from the sediment. The carbon amended treatment exhibited an overall net N 2 uptake, whereas the control receiving only aquaculture waste exhibited net N 2 production, suggesting that carbon supplementation enhanced nitrogen fixation. The higher NH + 4 efflux and N 2 uptake was further supported by meta-genome predictions that indicate that organic-carbon addition stimulated DNRA over denitrification. These findings indicate that carbon addition may potentially result in greater retention of nitrogen within the system; however, longer-term trials are necessary to determine whether this nitrogen retention is translated into improved sea cucumber biomass yields. Whether this truly constitutes a remediation process is open for debate as there remains the risk that any increased nitrogen retention may be temporary, with any subsequent release potentially raising the eutrophication risk. Longer and larger-scale trials are required before this approach may be validated with the complexities of the in-system nitrogen cycle being fully understood.