Iranian J Parasitol: Vol. 6, No.3, 2011, pp.89 -95 one Nosema species, N. apis, infected honey bees. The presence of the reports based on the incidence of colony depopulation and high prevalence of nosemosis, with different clinical and epidemiological pattern in Iran, noticed us that its cause may potentially be this addition species. The most significant difference between the two microsporidian, is how quickly N. ceranae can cause a col- ony to die. The foraging force to be affected the most. They leave the colony and are too weak to return, thus dying in the field. Regarding to the similarity of spores of the two Nosema species and it is hardly distin- guishing by optical microscopy, so that we use PCR technique in the diagnosis and identification of honeybee microsporidians. The aim of performing this study was to show the probability of presence of new spe- cies of Nosema in Iran using small- sub unit rRNA (16SrRNA). Based on molecular data, our studies confirm that the microsporidia isolated from Savadkouh samples of honey- bees are N. ceranae. Polymerase Chain Reaction with the N. cer- anae specific primers, 218MITOC were positive in all six samples sent to our de- partment for Nosema analysis, Subsequent PCR assays confirmed the suitability of our design. The N. ceranae SSUrRNA gene- specific primers used here successfully am- plified a 218-bp fragment from N.ceranae spores. Conversely, these primers did not amplify DNA preparations from uninfected honeybees. This is the first molecular survey on N. cer- anae in the country attempting to identify and characterize the species of this genus. However, this is the first study that provides evidence concerning the species of N. cer- anae, which causes severe disease among honeybees in Iran. It seems that the lack of seasonality detected and the increasing number of pathological samples sent to our laboratory in last years without any compatible Nosema signs of in- 93 fection can be related to the N. ceranae prevalence in other regions in Iran that needs to investigate. Some differences in the epi- demiology and pathology caused by this agent may explain this situation. According to earned results, we can not state the exact time of the beginning of infection, but we suggest its occurrence in this country. Over the past years, many beekeepers in the country have noticed a change in the course of nosemosis in their colonies, especially loss of honeybees the whole year around and quick die off colonies in the wintertime, N. apis, on the other hand, generally affects colonies during spring (25). It seems that these differences may be due to the emergent parasite, N. ceranae, as the relationship be- tween recent Iran honeybee colony losses and nosemosis. Analyses of more colonies with and without apparent Nosema disease are needed to de- termine the prevalence of this new agent in Iran. According to announcement of the many of the investigators, there is a relation- ship between N. ceranae and colony collapse disorder. It should be noticed that the other variety of reasons of the occurrence of dis- ease should not be ignored. Recent survey and experimental work suggest that N. cer- anae is a serious threat to the global bee- keeping industry (3, 5, 15). Intensive surveys are needed to determine the distribution and prevalence of N. ceranae in Iran. Acknowledgments This study was supported by Veterinary Or- ganization of Iran. We thank beekeepers in mention province for preparing the bees samples. The authors declare that there is no conflict of interests.