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Physiological processes and gross energy budget of the longline-cultured Pacific oyster Crassostrea gigas were investigated in Geoje–Hansan Bay, Korea during two entire culturing periods. Based on physiological measurements of food consumption, feces production, ammonium excretion, and respiration from July 2008 to February 2009 and from July 2013 to February 2014, scope for growth appeared to be positive during most of the culturing period, except for one period with extremely high temperatures (up to 25°C). Estimates of physiological energy production matched well with tissue energy increment measured by gross biochemical composition during the culturing period, suggesting that the oysters might adjust their physiological performance to relatively low concentrations of suspended particulate matter in the bay to optimize energy acquisition. Such an adaptive adjustment includes an increased absorption of energy and a reduced loss of metabolic and excretory energy, resulting in positive production under high culturing density. Using physiological measurements, we further assessed the feedback effects of the longline aquaculture of oysters on the bay system. Ecological efficiency, estimated by a series of energetic efficiencies at the whole bay level, was low compared with Lindeman’s law of trophic efficiency. Biodeposition and ammonia excretion rates in this study were relatively low compared with other intertidal plastic bag cultures. These results indicate that the cultured oysters might have only minor effects on benthic and pelagic environments of the bay. Overall, our results suggest that the adaptive physiological performance of oysters and consequently weak feedback effects on ambient habitats should facilitate sustainable longline aquaculture in the bay for a prolonged period without severe habitat deterioration.

Biological nitrogen fixation (BNF) by canonical molybdenum and complementary vanadium and iron-only nitrogenase isoforms is the primary natural source of newly fixed nitrogen. Understanding controls on global nitrogen cycling requires knowledge of the isoform responsible for environmental BNF. The isotopic acetylene reduction assay (ISARA), which measures carbon stable isotope (13C/12C) fractionation between ethylene and acetylene in acetylene reduction assays, is one of the few methods that can quantify isoform-specific BNF fluxes. Application of classical ISARA has been challenging because environmental BNF activity is often too low to generate sufficient ethylene for isotopic analyses. Here we describe a high sensitivity method to measure ethylene δ13C by in-line coupling of ethylene preconcentration to gas chromatography-combustion-isotope ratio mass spectrometry (EPCon-GC-C-IRMS). Ethylene requirements in samples with 10% v/v acetylene are reduced from > 500 to ~ 20 ppmv (~ 2 ppmv with prior offline acetylene removal). To increase robustness by reducing calibration error, single nitrogenase-isoform Azotobacter vinelandii mutants and environmental sample assays rely on a common acetylene source for ethylene production. Application of the Low BNF activity ISARA (LISARA) method to low nitrogen-fixing activity soils, leaf litter, decayed wood, cryptogams, and termites indicates complementary BNF in most sample types, calling for additional studies of isoform-specific BNF.

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