Objective: To clarify the taxonomic status of Anopheles yatsushiroensis in Rongcheng City of Shandong Province.
Methods: By the end of August 2009, Anopheles yatsushiroensis were collected in the western suburbs of Wangjia Village of Rongcheng City, and raised for the next generation of various types of adult mosquitoes. Through morphological identification of filial generation various types, mosquito feet of 1-2 mosquitoes of various types were taken for complete DNA extraction, and complete sequence analysis of rDNA-ITS2 alignment was made by PCR. Multiple sequnence comparison was carried out with those previously documented by DNAstar7.1, including An. yatsushiroensis in Shandong Province (YSD, AY306128), An. yatsushiroensis in Sichuan Province (YSC, AY170925), An. yatsushiroensis in Liaoning Province (YLN, AY170923) and An. yatsushiroensis from South Korea (YK, AF146749).
Results: 56 An. yatsushiroensis with blood meal were collected in Rongcheng City, of which 7 successfully laid eggs, and received 354 adult mosquitoes of filiar generation, including 240 An. yatsushiroensis (Y type), 8 An. pullus (P type) and 106 hybrid type (H). The rDNA-ITS2 sequence alignment homology between single-parent female offspring of adult mosquitoes and various types of An. yatsushiroensis segments (JN865249, JN865246, JN865247, JN865248) was 98.7-100%, and the rDNA-ITS2 sequence alignment homology to those from Sichuan (YSC), Liaoning (YLN) and Korea (YK) was 98.5%-99.3%, 98.7%-99.6% and 98.7 to 100%, respectively, while to the original An. yatsushiroensis from Rongcheng (YSD), it was only 67.1%-68.5%.
Conclusion: There are Y, P and H types of An. yatsushiroensis in Rongcheng City, Shandong Province, but the rDNA-ITS2 sequence is close to An. pullus. The local distribution of An. pullus is deduced.