<p>In order to investigate the effects of macrobenthos on the abundance and community composition of ammonia-oxidizing prokaryotes under different temperature conditions, laboratory microcosms containing two kinds of macrobenthos (<i>Corbicula fluminea</i> and <i>Tubificid</i> worms) were constructed. Real-time polymerase chain reaction (PCR) and clone libraries were applied to analyze the ammonia-oxidizing archaea (AOA) and bacteria (AOB) communities in the surface sediments. The lowest abundances of the archaeal and bacterial <i>amoA</i> gene were found in the samples cultured under 28 °C (archaeal <i>amoA</i> gene abundance, 2.71 × 10<sup>6</sup> copies/g dry sediment; bacterial <i>amoA</i> gene abundance, 1.17 × 10<sup>7</sup> copies/g dry sediment) of the <i>C. fluminea</i> group. However, there was no significant difference in terms of the abundance of archaeal <i>amoA</i> gene in all <i>Tubificid</i> worms treatment groups. Compared to the community composition of AOB, greater variations in the community composition of AOA were observed among the three different temperature groups.</p