Fecal Viral Diversity of Captive and Wild Tasmanian Devils Characterized Using Virion-Enriched Metagenomics and Metatranscriptomics
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The Tasmanian devil is an iconic Australian marsupial that has suffered an 80% population decline due to a contagious cancer, devil facial tumor disease, along with other threats. Until now, viral discovery in this species has been confined to one gammaherpesvirus (dasyurid herpesvirus 2 [DaHV-2]), for which captivity was identified as a significant risk factor. Our discovery of 24 novel marsupial-associated RNA and DNA viruses, and that viral diversity is lower in captive than in wild devils, has greatly expanded our knowledge of gut-associated viruses in devils and provides important baseline information that will contribute to the conservation and captive management of this endangered species. Our results also revealed that a combination of virion-enriched metagenomics and metatranscriptomics may be a more comprehensive approach for virome characterization than either method alone. Our results thus provide a springboard for continuous improvements in the way we study complex viral communities.
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This is the second time that RHDV2 has been found in non-rabbit hosts.
“This is the second time that RHDV2 has been found in non-rabbit hosts (Chong et al. 2019), presumably through rabbit consumption in this case.”
From Red fox viromes in urban and rural landscapes (2020) · cites this paper as Chong et al. 2019
The Results section reports PCR-confirmed RHDV2 in Tasmanian devil fecal samples, supporting detection in a non-rabbit animal. Our copy does not establish that the citing paper’s finding was the second such detection. The Discussion also cautions that the virus may have come from the devils’ diet rather than replicating in them; page numbers are unavailable in our copy. (LLM, conf=0.86)
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“36,45”
Ji, J., Hu, W., Liu, Q., Zuo, K.-J., Zhi, G.-L., Xu, X., Kan, Y.-C., Yao, L., & Xie, Q. (2020) · Frontiers in Veterinary Science · doi:10.3389/fvets.2020.580836 · Cited by 19
“Studies have reported that expanding its host spectrum of chaphamaparvoviruses, these viruses were also identified in mammals such as mice (5, 6), simian (7), swine (8), and Tasmanian devils (9).”
BackgroundChong, R., Cheng, Y., Hogg, C., & Belov, K. (2020) · Frontiers in Microbiology · doi:10.3389/fmicb.2020.01058 · Cited by 19 · see citing works
Self-citationCanuti, M., Verhoeven, J., Munro, H., Roul, S., Ojkić, D., Robertson, G., Whitney, H., Dufour, S., & Lang, A. S. (2021) · Viruses · doi:10.3390/v13020193 · Cited by 18 · see citing works
“These included in total 61 sequences from avian viruses and a sub-clade of three sequences identified in Tasmanian devils [41], while no other virus of avian origin was identified in other clades.”
BackgroundIdentification of Novel Astroviruses in the Gastrointestinal Tract of Domestic Cats
van Brussel, K., Wang, X., Shi, M., Carrai, M., Li, J., Martella, V., Beatty, J., Holmes, E., & Barrs, V. (2020) · Viruses · doi:10.3390/v12111301 · Cited by 16 · see citing works
“Viral nucleic acids were isolated from the faecal samples of all cats using a previously published protocol for viral particle enrichment [22] with minor modifications [23].”
MethodInfluentialSelf-citationHarvey, E., Mifsud, J. C. O., Holmes, E. C., & Mahar, J. (2023) · Virus Evolution · doi:10.1093/ve/vead061 · Cited by 15 · see citing works
“Two Tasmanian devil–associated papillomaviruses were previously identified in a faecal meta-transcriptome, but as these animals are carnivores, it was impossible to determine their true host (Chong et al. 2019).”
BackgroundResultInfluentialSelf-citationHuaman, J., Pacioni, C., Sarker, S., Doyle, M., Forsyth, D., Pople, A., Hampton, J., Carvalho, T. G., & Helbig, K. (2021) · Viruses · doi:10.3390/v13081492 · Cited by 15
“In Australia, PBVs have been previously detected by metagenomics in stool samples collected from humans [4], wild birds [7,8], European rabbits (Oryctolagus cuniculus) [9], and Tasmanian devils (Sarcophilus harrisii) [10].”
BackgroundRecent Progress in Torovirus Molecular Biology
Ujike, M., & Taguchi, F. (2021) · Viruses · doi:10.3390/v13030435 · Cited by 15 · see citing works
“full genome sequences of ToV from Tasmanian Devil [18] and goat have been reported.”
BackgroundInfluentialVirome Analysis of Normal and Growth Retardation Disease-Affected Macrobrachium rosenbergii
Zhou, D.-D., Liu, S., Guo, G., He, X.-Y., Xing, C., Miao, Q.-J., Chen, G., Chen, X.-L., Yan, H., Zeng, J., et al. (2022) · Microbiology spectrum · doi:10.1128/spectrum.01462-22 · Cited by 13
“show high tolerance to stochastic errors to the relative abundance of viromic taxa (2, 3).”
BackgroundChen, X.-X., Wu, W.-C., & Shi, M. (2021) · Viruses · doi:10.3390/v13061042 · Cited by 13
“Indeed, a study that compares metatranscriptomics and viral particle enrichment approaches [49–52] on the same set of samples revealed that the metatranscriptomics approach discovered less DNA viruses in general, and for those revealed by both approaches, the metatranscriptomics approach tended to reveal lower abundan…”
ResultSelf-citationHargitai, R., Boros, Á., Pankovics, P., Mátics, R., Altan, E., Delwart, E., & Reuter, G. (2020) · Archives of Virology · doi:10.1007/s00705-020-04862-6 · Cited by 13
Rabbit Hemorrhagic Disease Virus Isolated from Diseased Alpine Musk Deer (Moschus sifanicus)
Bao, S.-J., An, K., Liu, C.-G., Xing, X.-Y., Fu, X.-P., Xue, H.-W., Wen, F.-Q., He, X., & Wang, J.-F. (2020) · Viruses · doi:10.3390/v12080897 · Cited by 12 · see citing works
“Rabbits in the G1 group were inoculated intraperitoneally with 1 mL of 10% 0.22 µm syringe filtrated liver homogenate the same as the initial preparation used for SISPA and sequencing.”
BackgroundMethodInfluentialByrne, A., Marnell, F., Barrett, D., Reid, N., Hanna, R., Mcelroy, M., & Casey, M. (2022) · Pathogens · doi:10.3390/pathogens11030288 · Cited by 10 · see citing works
Discovery and Prevalence of Divergent RNA Viruses in European Field Voles and Rabbits
Tsoleridis, T., Chappell, J., Monchatre-Leroy, E., Umhang, G., Shi, M., Bennett, M., Tarlinton, R., McClure, C. P., Holmes, E., & Ball, J. (2019) · Viruses · doi:10.3390/v12010047 · Cited by 10
“Indeed, previous studies have shown the detection of viruses in gut and fecal samples of humans and animals that are likely derived from their diet [40] [41] [42] .”
BackgroundSelf-citationZhang, M., Zhou, Y., Cui, X., & Zhu, L. (2024) · Microorganisms · doi:10.3390/microorganisms12040713 · Cited by 8 · see citing works
“These findings further expand our understanding of gut phage variations across geographically distinct giant panda populations, providing valuable insights for the ecological health and conservation strategies tailored for these iconic species.”
BackgroundNovel polyomaviruses identified in fecal samples from four carnivore species
Kraberger, S., Serieys, L. E. K., Riley, S. P. D., Schmidlin, K., Newkirk, E. S., Squires, J. R., Buck, C., & Varsani, A. (2023) · Archives of Virology · doi:10.1007/s00705-022-05675-5 · Cited by 9 · see citing works
Identification of a novel polyomavirus from a marsupial host
Dunowska, M., Perrott, M., & Biggs, P. (2022) · Virus Evolution · doi:10.1093/ve/veac096 · Cited by 9 · see citing works
Kayigwe, A. N., Darby, J., Lyons, A., Patchett, A. L., Lisowski, L., Liu, G.-S., & Flies, A. S. (2022) · bioRxiv · doi:10.1101/2022.05.29.493930 · Cited by 9
Guo, G., Wang, M.-H., Zhou, D.-D., He, X., Han, P., Chen, G., Zeng, J., Liu, Z., Wu, Y., Weng, S., et al. (2023) · Microbiology spectrum · doi:10.1128/spectrum.01439-23 · Cited by 8
“stochastic error derived from the relative abundance of viromic taxa (7, 8).”
BackgroundFunctional Diversity within Gut Microbiomes: Implications for Conserving Biodiversity
Dodd, C. S., & Grueber, C. E. (2021) · Conservation · doi:10.3390/conservation1040024 · Cited by 10 · see citing works
“Studies in Tasmanian devils have shown that the gut microbiome hosts a rich diversity of viruses, which can play a key role in individual health and future conservation efforts [96].”
BackgroundSelf-citationZhao, X., Dai, C., Qian, S., Tang, Q., Li, L., Hao, Y.-J., Zhou, Z.-J., Ge, X.-Y., Gong, C., & Yuan, J.-F. (2023) · Microbiology spectrum · doi:10.1128/spectrum.00810-23 · Cited by 6 · see citing works
“Five fecal swabs were pooled as one sample for library preparing and sequencing (37, 38).”
Tokarz-Deptuła, B., Kulus, J., Baraniecki, Ł., Stosik, M., & Deptuła, W. (2024) · International Journal of Molecular Sciences · doi:10.3390/ijms25105342 · Cited by 5 · see citing works
“…genetic material of the original infectious agent, i.e., RHD virus (now GI.1–RHDV), was also found in the Tasmanian devil ( Sarcophilus harrisii ) [67], the Mediterranean mouse ( Mus spretus ), the scrub mouse ( Apodemus sylvaticus ) [68], the Mediterranean vole ( Microtus duodecimcostatus ) and…”
BackgroundCui, H., Zhang, Z.-B., Xu, X., Zuo, K.-J., Ji, J., Guo, G., Kan, Y.-C., Yao, L., Xie, Q.-M., & Bi, Y.-Z. (2023) · Frontiers in Veterinary Science · doi:10.3389/fvets.2023.1252628 · Cited by 5
Abundant dsRNA picobirnaviruses show little geographic or host association in terrestrial systems.
Knox, M., Wierenga, J., Biggs, P., Gedye, K., Almeida, V., Hall, R. J., Zikusoka, G. K., Rubanga, S., Ngabirano, A., Valdivia-Granda, W., et al. (2023) · Infection, Genetics and Evolution · doi:10.1016/j.meegid.2023.105456 · Cited by 5
Picobirnavirus: how do you find where it’s hiding?
Hutton, A., Grigson, S., Bartle, L., Papudeshi, B., Mallawaarachchi, V., Tarasenko, A., Mitchell, J. G., & Edwards, R. (2025) · Critical reviews in microbiology · doi:10.1080/1040841x.2025.2560918 · Cited by 4
“Similarly, chong et al. (2019) identified six novel PBv genomes of tasmanian devil feces, revealing high diversity and widespread phylogenetic dispersion of RdRp sequences.”
Ji, J., Mu, X., Pan, S., Xu, X., Zhang, S., Huang, H., Li, Y., Bi, Y.-Z., & Yao, L.-G. (2025) · Frontiers in Cellular and Infection Microbiology · doi:10.3389/fcimb.2025.1490948 · Cited by 4
Bacterial and Viral Diversity of Didelphid Opossums from Brazil
Caserta, L. C., do Nascimento, G. M., Joshi, L. R., Simão, R. M., Miller, M. E., Felippe, P. A. N., Diel, D., & Arns, C. W. (2023) · EcoHealth · doi:10.1007/s10393-023-01667-2 · Cited by 4
“Astroviruses, picobirnaviruses, parvoviruses and papillomaviruses were also detected in the fecal virome of Tasmanian devils (Rowena et al. 2019), where a large proportion of the viral contigs belonged to either nonver-tebrate eukaryotic viruses, including plant viruses, insect viruses, and…”
Li, N., Bai, Y., Yan, X., Guo, Z., Xiang, K., Yang, Z., Shangguan, H., Ge, J.-W., & Zhao, L. (2023) · Frontiers in Veterinary Science · doi:10.3389/fvets.2023.1233972 · Cited by 4 · see citing works
“Several studies have also detected chaphamaparvoviruses in mammals such as simians (6), Tasmanian devils (7), rat (8, 9), swine (10), and birds, including turkey (11), red-crowned crane (12), chicken (13), and peafowl (14).”
BackgroundPiewbang, C., Lohavicharn, P., Nguyen, T. V., Punyathi, P., Kasantikul, T., & Techangamsuwan, S. (2023) · Veterinary Quarterly · doi:10.1080/01652176.2023.2185696 · Cited by 4
Wang, X., Carrai, M., Brussel, K. V., Feng, S., Beatty, J. A., Shī, M., Holmes, E. C., Li, J., & Barrs, V. R. (2022) · Viruses · doi:10.3390/v14071412 · Cited by 5 · see citing works
“Viral nucleic acids (RNA and DNA) were extracted using the QIAamp Viral RNA Mini Kit (Qiagen, Hilden, Germany) after enrichment according to a published protocol [21] with minor modifications [22] and split into two aliquots.”
MethodSelf-citationFarming reshapes the gut resistome, virulome, and mobilome of Cervidae
Sun, Y.-Z., Su, J.-W., Elsheikha, H. M., Lou, W.-B., Song, Y.-H., Li, J.-M., Liu, F., Cai, R.-P., Leng, X., Gong, Q.-L., et al. (2026) · Virulence · doi:10.1080/21505594.2026.2728506 · Cited by 2 · see citing works
Guo, X., Zhang, Y., Pan, Y., Yang, K.-K., Tong, X., & Wang, Y. (2023) · Animals · doi:10.3390/ani13162617 · Cited by 2 · see citing works
“ChPV can infect various animals, including bats [3], rodents [4], birds/poultry [5–8], pigs [9], dogs [10], domestic cats [11], Tasmanian devils [12], wild mammals [13,14], and humans [15].”
BackgroundEvidence of Viral Communities in Three Species of Bats from Rural Environment in Mexico
Barrón-Rodríguez, R. J., Parra-Laca, R., Anaya, E. R., Romero-Espinoza, J. A., Ayala-Sumuano, J. T., Vázquez-Pérez, J. A., Garcı́a-Espinosa, G., & Rubio, E. L. (2022) · Acta Chiropterologica · doi:10.3161/15081109acc2022.24.1.020 · Cited by 2 · see citing works
Malik, Y., Bhat, S., Vlasova, A., Wang, F.-I., Touil, N., Ghosh, S., Dhama, K., Yadav, M., & Singh, R. (2020) · Emerging and Transboundary Animal Viruses · doi:10.1007/978-981-15-0402-0_14 · Cited by 2
Empowering One Health with metagenomics
Harvey, E., van Brussel, K., & Holmes, E. C. (2026) · One Health Outlook · doi:10.1186/s42522-026-00225-4 · Cited by 1
“In contrast, direct comparisons of virion-enriched metagenomics and metatranscriptomics (i.e. total RNA sequencing) with rRNA depletion have shown that while virion enrichment performed better for the detection of DNA viruses, metatranscriptomics was superior overall [31].”
Self-citationThe final frontier: using carcasses for one health surveillance at the ecosystem interface
Barton, K. A., Finnerty, P., Rupasinghe, R., González-Crespo, C., Mahar, J. E., Eden, J.-S., Meisuria, N. Y., Martínez-López, B., Newsome, T. M., Peel, A. J., et al. (2025) · Frontiers in Veterinary Science · doi:10.3389/fvets.2025.1683110 · Cited by 1 · see citing works
French, R. K., Pichlmueller, F., Waller, S. J., Dubrulle, J., Tuxford, J., Veale, A., & Geoghegan, J. (2025) · bioRxiv · doi:10.1128/jvi.01440-25 · Cited by 1 · see citing works
“It also had the ribosomal frameshift (AAAAAAC) near the end of ORF1a, to create ORF1a/b. Possum astrovirus was most closely related to Tasmanian devil - associated astrovirus 1 (28), with 56% amino acid sequence similarity.”
Smith, I. L., Huang, N., Pavy, M., Gofton, A., Fahri, O., Kardia, E., Mourant, R., Chong, S., Jenckel, M., Hall, R., et al. (2025) · Wildlife research (East Melbourne) · doi:10.1071/wr24122 · Cited by 1
TaqMan-based real-time polymerase chain reaction for the detection of feline chaphamaparvovirus
Li, S., Huo, X., Mu, Y.-Y., Liu, X., Wu, J.-Y., Chen, Y., & Wang, Y. (2024) · 3 Biotech · doi:10.1007/s13205-024-03917-8 · Cited by 1
“However, it was found to infect other vertebrates (such as bats, rats, mice, apes, pigs, birds, tur-keys, peacocks, chickens and Tasmanian devils) (Chong et al. 2019; Fahsbender et al. 2019; Kapusinszky et al. 2017; Li et al. 2020; Palinski et al. 2016; Lima et al. 2019; Williams et al. 2018; Yang…”
Circulation of picobirnavirus in Neotropical free-ranging mammals
Vanderhoeven, E., Paresque, R., Mello, I. O., Nates, S., Vicentini, F., Díaz, A., & Masachessi, G. (2023) · Veterinary research communications · doi:10.1007/s11259-023-10282-z · Cited by 1
“Furthermore, using metagenomics, the presence of multiple PBVs has been reported in studies exploring viromes in the stool (Chong et al. 2019; Mahar et al. 2020) and plasma (Li et al. 2015) obtained from mammals.”
Metagenomic and Molecular Detection of Novel Fecal Viruses in Free-Ranging Agile Wallabies
Okoh, G. R., Ariel, E., Whitmore, D., & Horwood, P. F. (2023) · EcoHealth · doi:10.1007/s10393-023-01659-2 · Cited by 1 · see citing works
“However, both approaches failed to detect all of the viruses, especially those present in low amount (Chong et al., 2019).”
Red fox viromes across an urban-rural gradientPreprint
Campbell, S. J., Ashley, W., Gil-Fernández, M., Newsome, T., Giallonardo, F. D., Ortiz-Baez, A., Mahar, J., Towerton, A. L., Gillings, M., Holmes, E., et al. (2020) · bioRxiv · doi:10.1101/2020.06.15.153858 · Cited by 1 · see citing works
“This is the second time 273 that RHDV2 has been found in non-rabbit hosts (42), presumably through rabbit consumption in this case.”
BackgroundSelf-citationJolly, C., Lee, Q., Padula, M., Pinello, N., Williams, S., O'Rourke, M. B., Fumagalli, M., Orkin, J. D., Shaban, B., Brenner, O., et al. (2019) · bioRxiv · doi:10.1101/732537 · Cited by 1
Fan, S., Liu, H., Yan, Y., Xu, M., Wan, X., Hao, Y., Gong, C., Wang, C., Zhang, Y., Liu, D., et al. (2026) · Animal Microbiome · doi:10.1186/s42523-026-00528-x
“Lifestyles could give rise to differences in host-associated virome [85], and differences in the virome among individuals exposed to identical environmental conditions may arise from individual variability or other unidentified factors.”
A molecular inventory of the faecal microbiomes of 23 marsupial species
Bowerman, K., Soo, R. M., Chaumeil, P., Blyton, M. D. J., Sørensen, M., Gunbilig, D., Malig, M., Islam, M., Zaugg, J., Wood, D., et al. (2025) · bioRxiv · doi:10.1099/mgen.0.001601
“…culture-independent methods, and only 12 of these have been analysed using metagenomics (Table S1, available in the online Supplementary Material) To our knowledge, no other omic techniques have been applied to marsupial gut microbiomes, and marsupial gut viruses remain largely unexplored [30, 31].”
Rodents as potential reservoirs for toroviruses
Buigues, J., Viñals, A., Martínez-Recio, R., Monrós, J. S., Sanjuán, R., & Cuevas, J. M. (2025) · bioRxiv · doi:10.1186/s12985-025-02933-1
“…Article 2), procedures used in this study (i.e. capture, non-invasive handling and in situ release of wild animals) are not subject to the condition of animal experimentation and, therefore, an IACUC approval document is not required, but specifically a permit from the competent regional authority.”
Shao, Y., Liu, Y., Huang, D., Wang, Q., He, X.-X., Zhao, W., Zhao, Y., Ma, H., Xing, X.-Y., Zhang, Z.-X., et al. (2024) · Pathogens · doi:10.3390/pathogens13110925
“RHDV is believed to have very strict host specificity, and there is no evidence that the virus can infect animals other than rabbits either naturally or experimentally, nor is it known to cause disease in animals other than rabbits [13–16].”
Fecal Virome of Coendou spinosus (Paraguaian Hairy Dwarf Porcupine)
Miranda, T. D. S., de Matos, I. G., D'Arc, M., Moreira, F., Schiffler, F., Cosentino, M., Coimbra, A., de Oliveira, T. D., Mouta, R., Medeiros, G., et al. (2023) · bioRxiv · doi:10.1101/2023.09.24.559146
“Most were bacteriophages such as Siphoviridae , Myoviridae , Podoviridae , Autographiviridae , Schitoviridae and Demerecviridae, commonly found in the feces of animals and humans (Chong et al., 2019; Liang & Bushman, 2021; Ning et al., 2021) ( Supplementary Table S2 ) .”
Harvey, E., Mifsud, J. C. O., Holmes, E., & Mahar, J. (2023) · bioRxiv · doi:10.1101/2023.06.27.546737
Self-citationMultiple known and a novel parvovirus associated with an outbreak of feline diarrhea and vomitingPreprint
Li, Y., Gordon, E., Idle, A., Altan, E., Seguin, M., Estrada, M., Deng, X.-T., & Delwart, E. (2020) · bioRxiv · doi:10.1101/2020.03.24.005876
- Authors
- Chong, R., Shi, M., Grueber, C. E., Holmes, E. C., Hogg, C. J., Belov, K., & Barrs, V. R.
- Affiliations
- The University of Sydney (AU)
- Marie Bashir Institute for Infectious Diseases and Biosecurity, Sydney Medical School, University of Sydney, Sydney, New South Wales, Australia
- Corresponding author
- Barrs, Vanessa R.
- Venue
- Journal of Virology
- Publisher
- American Society for Microbiology
- Published
- 2019-06
- Volume
- 93
- Issue
- 11
- ISSN
- 0022-538X (ISSN-L)
- 1098-5514
- PMC
- PMC6532096
- OpenAlex
- W2921204261
- Cited by
- 77
- References
- 61
- Citations by year
- 2026: 3 · 2025: 8 · 2024: 7 · 2023: 16 · 2022: 12 · 2021: 10 · 2020: 18 · 2019: 3
- FWCI
- 8.75
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Virology (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- Department of Education and Training | Australian Research Council (LP140100508, FL1701000022)
- Topics
- Life Sciences › Agricultural and Biological Sciences › Animal Science and Zoology › Animal Virus Infections Studies
- Physical Sciences › Environmental Science › Ecology › Bacteriophages and microbial interactions
- Health Sciences › Medicine › Public Health, Environmental and Occupational Health › Zoonotic diseases and public health
- Keywords
- Tasmanian devil, fecal virome, captive vs wild populations, metatranscriptomics, devil facial tumour disease, endangered species conservation, captive management
- MeSH terms
- Animals, Animals, Wild, Animals, Zoo, Australia, Feces, Marsupialia, Virion, Gene Expression Profiling, Metagenomics, Endangered Species, Transcriptome
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