This study determined the spatial and seasonal density, number of genera, genera composition, maturity index and trophic structure of free-living nematode assemblages in the subtidal waters of the northern Beibu Gulf, South China Sea, and explored whether these five biotic characteristics were related to various environmental variables. Based on the data derived from samples collected seasonally at nine stations, the mean densities of nematodes decreased from alongshore to offshore station in the northern Beibu Gulf. However, the number of nematode genera increased from alongshore to offshore station. Non-parametric multidimensional scaling analysis showed no clear seasonal changes for nematode assemblages in most sampling stations. Higher densities of the genera Elzalia and Tricoma were found in offshore sampling stations, and a higher density of the genus Cheironchus was found in alongshore sampling stations. The mean percentage of each feeding type compared to the total numbers was highest in epigrowth feeders (2A), second highest in non-selective deposit feeders (1B), third highest in predators (2B), and lowest in selective deposit feeders (1A). There were significant negative correlations between nematode density and water depth and temperature; significant positive correlations between the number of nematode genera and water depth and salinity; and significant negative correlation between the maturity index of the nematode assemblage and organic matter. BIOENV analysis indicated that water depth, salinity, pH, median sediment particle size and organic matter were the most correlated combination of environmental variables affecting the nematode assemblages.