Ticks, as significant disease-transmitting vectors, have a profound impact on public health. Notably, Haemaphysalis japonica and Haemaphysalis megaspinosa, two hard tick species widely distributed across Asia and Russia, attract attention. However, their taxonomy and genetic differentiation have posed challenges. Previous studies struggled to differentiate between H. japonica and H. megaspinosa due to a lack of sequence data in the public database. This project was aimed to clarify the difference in microbiome composition between these two closely related tick species. The V3-4 region of the 16S rDNA was amplified using universal primers and the resulting PCR products were analyzed on an Illumina MiSeq platform.