D3184: Exploring tick "mycobiome" by using nested PCR targeting fungal ITS region

Ticks are known as reservoirs and vectors of numerous causative agents of infectious diseases of humans and domesticated animals. However, exploring the presence of pathogenic microorganisms in ticks revealed the presence of a diversity of non-pathogenic microorganisms, known as the tick microbiome. Studies of tick microbiome are mostly based on next-generation sequencing platforms that use universal 16S rRNA primers to amplify and identify sequences originating from genetic material extracted from ticks. In this way, numerous endosymbiotic bacteria are identified in ticks, but other types of microorganisms cannot be detected. The purpose of this study is to identify endosymbiotic fungi present in ticks – tick “mycobiome”. We used fungi-specific primers, that target the ITS region, in nested PCR. As a template we used DNA extracted from four tick species, including Ixodes ricinus, Dermacentor variabilis, Amblyomma americanum, and Amblyomma maculatum. Amplified tick sequences were submitted for sequencing and analyzed by BLAST in GenBank. Preliminary results show similarity to fungal species in the Ascomycota phylum. To obtain more precise data, amplified sequences are cloned into a pGEM-T vector, and analysis of selected clones is ongoing.