Stratified estuaries are home to expanding aquaculture activities whose ecological footprints can be observed through trends in microbial community respiration in the water column. Bottle incubations are widely used to measure water column community respiration in marine and freshwater ecosystems by measuring the flux of dissolved oxygen occurring in the bottle over a period of time. When in situ dissolved oxygen (DO) concentrations are markedly different than DO concentration of the incubation medium the potential for diffusion of oxygen across the bottle opening is great and may be especially pronounced in strongly stratified systems with relatively low rates of pelagic oxygen consumption. We incubated 60 Biochemical Oxygen Demand (BOD) bottles filled with sterilized water with DO concentrations ranging from 2.51 mg O2 L-1 to 10.03 mg O2 L-1 for 24 hours in a temperature controlled water bath. There was a significant relationship when DO flux was set as a function of initial DO (DO Flux = -0.0017x + 0.0085, r2 = 0.72, p < 2.2 e-16). DO fluxes ranged from -0.012 mg O2 L-1 hour-1 to 0.005 mg O2 L-1 hour-1 for bottles incubated with initial DO ranging from 10.03 mg O2 L-1 to 3.31 mg O2 L-1, respectively. These results suggest that diffusion across the ground glass seal of BOD bottles is possible and that extra precaution through parallel diffusion controls should be considered when measuring pelagic respiration using BOD bottle incubations in systems with relatively low or relatively high in situ DO concentrations.