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Viruses that infect non-Plasmodium spp. disease-causing protozoa affect the pathogen life cycle and disease outcomes. However, only one RNA virus (Matryoshka RNA virus 1) has been identified in Plasmodium, and none have been identified in zoonotic Plasmodium species. The rapid expansion of the known RNA virosphere via metagenomic sequencing suggests that this dearth is due to the divergent nature of RNA viruses that infect protozoa. We leveraged newly uncovered data sets to explore the virome of human-infecting Plasmodium species collected in Sabah, east (Borneo) Malaysia. From this, we identified a highly divergent RNA virus in two human-infecting P. knowlesi isolates that is related to the unclassified group \u2018ormycoviruses\u2019. By characterizing 15 additional ormycoviruses identified in the transcriptomes of arthropods, we show that this group of viruses exhibits a complex ecology as noninfecting passengers at the arthropod\u2013mammal interface. With the addition of viral diversity discovered using the artificial intelligence\u2013based analysis of metagenomic data, we also demonstrate that the ormycoviruses are part of a diverse and unclassified viral taxon. This is the first observation of an RNA virus in a zoonotic Plasmodium species. By linking small-scale experimental data to advances in large-scale virus discovery, we characterize the diversity and confirm the putative genomic architecture of an unclassified viral taxon. This approach can be used to further explore the virome of disease-causing Apicomplexa and better understand how protozoa-infecting viruses may affect parasite fitness, pathobiology, and treatment outcomes.<\/jats:p>","DOI":"10.1093\/ve\/veae091","type":"journal-article","created":{"date-parts":[[2024,11,6]],"date-time":"2024-11-06T09:03:12Z","timestamp":1730883792000},"source":"Crossref","is-referenced-by-count":4,"title":["A virus associated with the zoonotic pathogen <i>Plasmodium knowlesi<\/i> causing human malaria is a member of a diverse and unclassified viral taxon"],"prefix":"10.1093","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3663-7535","authenticated-orcid":false,"given":"Mary E","family":"Petrone","sequence":"first","affiliation":[{"name":"Sydney Infectious Diseases Institute, School of Medical Sciences, The University of Sydney , Sydney, NSW 2006,","place":["Australia"]},{"name":"Laboratory of Data Discovery for Health Limited , 19 Science Park West Avenue, Hong Kong Science Park, Pak Shek Kok, New Territories, Hong Kong SAR,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Justine","family":"Charon","sequence":"additional","affiliation":[{"name":"Fruit Biology and Pathology Unit, University of Bordeaux, INRAE , 71 Av. Edouard Bourlaux, Villenave-d\u2019Ornon, Bordeaux 33140,","place":["France"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew J","family":"Grigg","sequence":"additional","affiliation":[{"name":"Global and Tropical Health Division, Menzies School of Health Research, Charles Darwin University , John Mathews Building (Bldg 58), Royal Darwin Hospital Campus, Rocklands Drv., Casuarina, Darwin, NT 8010,","place":["Australia"]},{"name":"Infectious Diseases Society Kota Kinabalu Sabah-Menzies School of Health Research Clinical Research Unit , Kota Kinabalu, Sabah 88200,","place":["Malaysia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Timothy","family":"William","sequence":"additional","affiliation":[{"name":"Infectious Diseases Society Kota Kinabalu Sabah-Menzies School of Health Research Clinical Research Unit , Kota Kinabalu, Sabah 88200,","place":["Malaysia"]},{"name":"Subang Jaya Medical Centre , No. 1, Jalan SS12\/1A, Ss 12, Subang Jaya, Selangor 47500,","place":["Malaysia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8123-476X","authenticated-orcid":false,"given":"Giri S","family":"Rajahram","sequence":"additional","affiliation":[{"name":"Infectious Diseases Society Kota Kinabalu Sabah-Menzies School of Health Research Clinical Research Unit , Kota Kinabalu, Sabah 88200,","place":["Malaysia"]},{"name":"Queen Elizabeth Hospital II, Ministry of Health Malaysia , Lorong Bersatu, Off, Jalan Damai, Luyang Commercial Centre, Kota Kinabalu, Sabah 88300,","place":["Malaysia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jacob","family":"Westaway","sequence":"additional","affiliation":[{"name":"Global and Tropical Health Division, Menzies School of Health Research, Charles Darwin University , John Mathews Building (Bldg 58), Royal Darwin Hospital Campus, Rocklands Drv., Casuarina, Darwin, NT 8010,","place":["Australia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kim A","family":"Piera","sequence":"additional","affiliation":[{"name":"Global and Tropical Health Division, Menzies School of Health Research, Charles Darwin University , John Mathews Building (Bldg 58), Royal Darwin Hospital Campus, Rocklands Drv., Casuarina, Darwin, NT 8010,","place":["Australia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6154-4437","authenticated-orcid":false,"given":"Mang","family":"Shi","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Biocontrol, School of Medicine, Shenzhen Campus of Sun Yat-sen University, Sun Yat-sen University , Shenzhen 518107,","place":["China"]},{"name":"National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, Sun Yat-sen University , Shenzhen 518063,","place":["China"]},{"name":"Shenzhen Key Laboratory for Systems Medicine in Inflammatory Diseases, Shenzhen Campus of Sun Yat-sen University, Sun Yat-sen University , Shenzhen 518063,","place":["China"]},{"name":"Guangdong Provincial Center for Disease Control and Prevention , Guangzhou 510642,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicholas M","family":"Anstey","sequence":"additional","affiliation":[{"name":"Global and Tropical Health Division, Menzies School of Health Research, Charles Darwin University , John Mathews Building (Bldg 58), Royal Darwin Hospital Campus, Rocklands Drv., Casuarina, Darwin, NT 8010,","place":["Australia"]},{"name":"Infectious Diseases Society Kota Kinabalu Sabah-Menzies School of Health Research Clinical Research Unit , Kota Kinabalu, Sabah 88200,","place":["Malaysia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9596-3552","authenticated-orcid":false,"given":"Edward C","family":"Holmes","sequence":"additional","affiliation":[{"name":"Laboratory of 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