Nosemosis is the most common disease in adult bees. Nosema apis and Nosema ceranae species are agents of important economic losses to beekeepers around the world. The severity of disease at various area is different. Previously, N. apis was observed in areas with a long winter, especially in late winter and early spring. But in recent years, disease has been reported in the warm seasons. The studies indicated that a new species as N. ceranae is involvement in loss and mortality in adult bees. Therefore, diagnosis and differentiation of Nosema species is importance at colony collapse disorders (CCD). The aim of this Research was a retrospective study on Nosema samples isolated from apiaries. Forty- one Nosema Sp. Positive samples were collected from five provinces during 2004 to 2013. The samples were tested by multiplex PCR method using both primers of N. ceranea and N. apis were simultaneously. All of samples were positive for N. ceranea. The products were sent for sequencing. The results show that N. ceranea has spread in Iran, from previous years almost simultaneously with other parts of the world.