To identify the key factors influencing the trimethylamine N-oxide (TMAO) content of teleost fishes living in shallow seas and the epipelagic zone of the deep ocean, the muscle TMAO content was measured in 152 teleost fishes (21 species) collected from the marginal seas of China and the epipelagic zone of the northwest Pacific Ocean (NWPO) during May-July 2021. The results showed that the TMAO content in all fishes varied from 4.99 to 82.97 mmol kg, and it varied notably among different species, e.g., the highest average content (72.71 ± 8.22 mmol kg in Argyrosomus argentatus) was 1 order of magnitude higher than the lowest one (Scomber japonicus), but the ratios of the highest content to the lowest content in each species varied from 1.29 to 3.28, suggesting that the interspecific variations in TMAO content were obviously greater than the intraspecific variations. Moreover, no correlation was observed between the TMAO content of the 152 fishes and the corresponding environmental factors of seawater depth, salinity and temperature, indicating that species played a more important role than environmental factors in driving TMAO accumulation. To exclude the influence of species, intraspecies correlations between TMAO content and environmental factors were analyzed. In the marginal seas of China, only ∼8 % of the TMAO content of teleost fishes (1 species) showed a positive correlation with salinity and depth, but ∼50 % of the TMAO content (5 species) was negatively correlated with temperature. Moreover, the TMAO content of the fish increased by 4.66 ± 1.38 % compared with their corresponding intraspecific average values for every 1 °C of temperature decrease. A similar phenomenon was also found in the TMAO content of pelagic teleost fishes in the NWPO, suggesting that temperature was a key environmental factor affecting the TMAO content of teleost fishes in shallow seas and the epipelagic zone of the deep ocean.