Macroinvertebrate assemblages and its association with environmental factors at the 11 artificial subtropical ponds of Taiwan were examined using the multivariate analysis software STATICO. The aims of the study were to determine whether spatial and seasonal variation of macroinvertebrate assemblages changed seasonally, to examine which environmental factors determined the spatial and temporal structure of maroinvertebrate assemblages, and to compare between-pond variations in the taxon composition of macroinvertebrates. Macroinvertebrates were collected seasonally by a corer and a sweep net in 2007, and 13 physical and chemical factors were measured at the same time. A total of 31 macroinvertebrate taxa were collected during the sampling period, and the most dominant taxa were Chironomidae (31.7% of total animal abundance) and Tubificidae (22.4%). STATICO identified pond size, pond depth, sediment depth, and altitude as the major abiotic factors and Bufo melanostictus (Amphibia) as the major biotic factor to influence macroinvertebrate assemblages at these ponds. These factors changed with seasonality. For example, the abundance of B. melanostictus was the most important factor during the spring but became much less important in other seasons. According to the spatial distribution patterns of macroinvertebrate assemblages, macroinvertebrates could be split into two groups based on their dispersal. The active dispersers, such as insect taxa, were strongly associated with pond size and the passive dispersers, such as non-insect taxa, were strongly associated with the pond depth and/or sediment depth. The results of this study suggested that pond size might influence macroinvertebrate assemblages through their dispersal mechanisms and that the environmental factors which influenced the macroinvertebrate assemblages most changed with seasons in this study area.