Integrated Omics Reveal the Pathogenic Potential of Blastocystis sp. ST2

Blastocystis sp. is a zoonotic unicellular eukaryote that is distributed worldwide. The pathogenicity of Blastocystis sp. has been debated over the years. In this study, mice were infected with Blastocystis sp. ST2 to assess the impact and underlying mechanisms on the host by integrating transcriptomics, metabolomics, and gut microbiomes. Transcriptomic analysis revealed significant differences in the expression of genes related to inflammatory cytokines, tumors, and neuropathic disease-related factors in mice infected with the parasite. A total of 430 differentially expressed genes (DEGs) were identified in Blastocystis-infected female mice, as compared with the control mice, and among these genes, the expression levels of 195 were upregulated (P  < 0.001), and that of 235 were downregulated (P  < 0.001). Similarly, there were different 478 DEGs in male mice, among which the expression levels of 122 genes (P  < 0.001) were upregulated, and that of 356 genes were downregulated (P  < 0.001). Kyoto encyclopedia of genes and genome analysis showed that 22 pathways in females and 28 pathways in males were enriched. Metabolomics results showed obvious metabolite changes in all mice infected with the parasite. In females, 82 different metabolites were identified, among which the expression levels of 27 metabolites were upregulated, and that of 55 metabolites were downregulated. In males, 118 metabolites were identified, among which the expression levels of 24 metabolites were upregulated, and that of 94 metabolites were downregulated. Microbiome analysis showed differences in the richness of bacterial families in Blastocystis sp. ST2-infected mice. LEfSe analysis showed differences in the abundance of bacterial families in female and male mice compared to the control groups. Multiomics analysis showed that the transcriptome, metabolome, and microbiome are interrelated. These results emphasize that Blastocystis sp. ST2 can negatively affect the host and may be a disease risk factor. The results provide insight into the mechanism of Blastocystis sp.-host interactions.