Abstract In this work, the TMAO-reduction by Shewanella baltica, one of the representative spoilage organisms in modified atmosphere packaged marine fish fillets, and the effect of acetate and CO 2 on this reduction were studied in vitro. The growth of S. baltica and the corresponding evolution of some compounds (acetate, lactate, pyruvate, glucose and trimethylamine (TMA)) were followed during storage at 4°C in two types of broths. The first medium was a defined medium (pH=6.8) to which lactate or pyruvate was added as hydrogen donor. Pyruvate showed to be more efficient as H-donor for S. baltica than lactate, as growth was much faster when equimolar amounts of pyruvate instead of lactate were present. Although the growth of S. baltica, when pyruvate is used as H-donor and no acetate is added, was not much inhibited by the CO 2-atmosphere, CO 2 had a pronounced effect on the studied reactions as it partly inhibited the reduction of pyruvate to acetate. The effect of acetate on this reaction was, on the other hand, not significant. To simulate the reactions occurring in situ, a buffered fish extract (pH=6.8) was used. In spite of the neutral pH, the growth of S. baltica in this medium was highly inhibited by relatively small concentrations of acetate (<0.3%). When 0.1% of acetate was added to the fish extract, less acetate was formed and lactate was more slowly consumed in comparison to the experiments without the addition of acetate. The consumption of lactate and the production of acetate were almost completely inhibited when the fish extract contained 0.25% of acetate. Apparently, the addition of acetate inhibited the use of lactate as H-donor. After extended storage times (17 days at 4°C) TMA production started. Most probably, alternative H-donors were used by S. baltica, from which the pathway seems to be less energy efficient. This can be deduced from the exceptional growth inhibition of S. baltica by small amounts of acetate. However, when practical storage times for fish (e.g. 6 days at 4°C after packaging) are considered, growth and TMAO-reduction by S. baltica was completely inhibited during this period by 0.25% of acetate.