To proper control the backwashing process of biological activated carbon (BAC) reactor and improve the overall operation performance, the evaluative indexes such as backwashing wastewater turbidity, organic pollutants removal rate of pre and post-backwashing, and the variation of biomass and biological activity in carbon column are used to compare and analyze the effect of three different combined water and air backwashing methods on the operation of BAC reactor. The result shows that intermittent combined water and air backwashing method is most suitable to BAC reactor. The biological activaty obviously increases by 62.5% after intermittent combined water and air backwashing process. While, the biological activaty using the backwashing method of air plus water and the backwashing method of water and air compounded plus water washing increases by 55.6%, 38.5%, respectively. After backwashing 308h, the reactor recovered to its normal function after intermittent combined water and air backwashing process with the removal rate of UV254 reaching to 60.0%. The fulvic-like fluorescence peak of backwashing water are very weak, and are characterized by low-excitation wavelength tryptophan like (peak S) and high excitation wavelength of tryptophan (peak T), which are caused by the microbial debris washed down. The three-dimensional fluorescence spectra also show that microbial fragments are easy to be washed clean with intermittent combined water and air backwashing.