The quantity of soil microbes and the structure of ammonium oxidizing bacterial (AOB) community were analyzed using the dilution plate counting and most probable number method (MPN), and denaturing gradient gel electrophoresis (DGGE), respectively. Fertilizer application tended to increase the number of soil microbes and alter the AOB community compared to the control with no fertilizer application (CK). Among the eight fertilizer treatments, soil samples from the treatments of mineral fertilizers (e.g., N, P, K) in combination with farmyard manure (M) had greater numbers of soil microbes and more complex structure of AOB community than those receiving mineral fertilizers alone. The principal component analyses (PCA) for ammonium oxidizing bacterial community structure showed that the eight fertilizer treatments could be divided into two PCA groups (PCA1 and PCA2). For the soil sampled after rice harvest, PCA1 included NP, NM, NPM and NPKM fertilizer treatments, while PCA2 was consisted of CK, N, M and NPK fertilizer treatments. For soil samples collected after wheat harvest, PCA1 was consisted of M, NM, NPM and NPKM fertilizer treatments, while PCA2 was composed of CK, N, NP and NPK fertilizer treatments. For a given rotation, the richness of AOB community in PCA1 was greater than that in PCA2. In addition, AOB community structure was more complex in the soil after rice harvest than that after wheat harvest. The results indicated that different fertilizer treatments resulted in substantial changes of soil microbe number and AOB community. Furthermore, mineral fertilizers (N, NP, NPK) combined with farmyard manure were effective for increasing the quantity of soil microbes, enriching AOB community, and improving the soil biofertility.