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We analysed whole\u2010genome sequences of CRC primary tumours, their corresponding metastases and matched normal tissue for sequences of viral, phage and bacterial species. Bacteriome analysis showed <jats:italic>Fusobacterium\u2009nucleatum<\/jats:italic>, <jats:italic>Streptococcus\u2009sanguinis<\/jats:italic>, <jats:italic>F.\u2009Hwasookii<\/jats:italic>, <jats:italic>Anaerococcus\u2009mediterraneensis<\/jats:italic> and further species enriched in primary CRCs. The primary CRC of one patient was enriched for <jats:italic>F.\u2009alocis<\/jats:italic>, <jats:italic>S.\u2009anginosus, Parvimonas\u2009micra<\/jats:italic> and <jats:italic>Gemella<\/jats:italic> sp. 948. Enrichment of <jats:italic>Escherichia\u2009coli<\/jats:italic> strains IAI1, SE11, K\u201012 and M8 was observed in metastases together with coliphages enterobacteria phage \u03c680 and Escherichia phage VT2\u03c6_272. Virome analysis showed that phages were the most preponderant viral species (46%), the main families being <jats:italic>Myoviridae<\/jats:italic>, <jats:italic>Siphoviridae<\/jats:italic> and <jats:italic>Podoviridae<\/jats:italic>. Primary CRCs were enriched for bacteriophages, showing five phages (Enterobacteria, Bacillus, Proteus, Streptococcus phages) together with their pathogenic hosts in contrast to normal tissues. The most frequently detected, and Blast\u2010confirmed, viruses included human endogenous retrovirus K113, human herpesviruses 7 and 6B, Megavirus chilensis, cytomegalovirus (CMV) and Epstein\u2013Barr virus (EBV), with one patient showing EBV enrichment in primary tumour and metastases. EBV was PCR\u2010validated in 80 pairs of CRC primary tumour and their corresponding normal tissues; in 21 of these pairs (26.3%), it was detectable in primary tumours only. The number of viral species was increased and bacterial species decreased in CRCs compared with normal tissues, and we could discriminate primary CRCs from metastases and normal tissues by applying the Hutcheson <jats:italic>t<\/jats:italic>\u2010test on the Shannon indices based on viral and bacterial species. Taken together, our results descriptively support hypotheses on microorganisms as potential (co)risk factors of CRC and extend putative suggestions on critical microbiome species in CRC metastasis.<\/jats:p>","DOI":"10.1002\/1878-0261.13070","type":"journal-article","created":{"date-parts":[[2021,7,30]],"date-time":"2021-07-30T15:39:56Z","timestamp":1627659596000},"page":"3363-3384","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":34,"title":["Metagenomic analysis of primary colorectal carcinomas and their metastases identifies potential microbial risk factors"],"prefix":"10.1002","volume":"15","author":[{"given":"Luigi","family":"Marongiu","sequence":"first","affiliation":[{"name":"Department of Experimental Surgery \u2013 Cancer Metastasis Medical Faculty Mannheim Ruprecht\u2010Karls University of Heidelberg Mannheim Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jonathan J. 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