Viral metagenomic analysis of the upper respiratory tract virome and identification of novel viruses in giant pandas

Zidan Zeng , Hongwen Zhang , Shiyin Huang , Yunli Li , Yue Chen , Shixing Yang , Quan Shen , Xiaochun Wang , Yuwei Liu , Ping Wu , Likai Ji , Songrui Liu , Wen Zhang

Animal Diseases ›› 2026, Vol. 6 ›› Issue (1) : 20

PDF
Animal Diseases ›› 2026, Vol. 6 ›› Issue (1) :20 DOI: 10.1186/s44149-026-00229-7
Short Communication
brief-report
Viral metagenomic analysis of the upper respiratory tract virome and identification of novel viruses in giant pandas
Author information +
History +
PDF

Abstract

The giant panda (Ailuropoda melanoleuca) is a flagship species for biodiversity conservation, yet the viral communities inhabiting its upper respiratory tract (URT) remain poorly characterized. In this study, we performed viral metagenomic sequencing on 13 pooled libraries derived from 130 nasopharyngeal swabs collected from giant pandas between 2018 and 2021. The assembly yielded 16 complete or near-complete viral genomes, predominantly of DNA viruses belonging to the families Papillomaviridae, Genomoviridae, and Parvoviridae. Notably, we identified the complete genome of a novel Parvoviridae species within the subfamily Densovirinae. Phylogenetic analysis revealed that this virus clusters with invertebrate-infecting viruses, suggesting that it likely represents an arthropod-associated viral element derived from respiratory parasites rather than a direct vertebrate pathogen. Although the alpha diversity remained stable, beta diversity analysis revealed significant temporal shifts in viral community composition (P = 0.02). This study provides the first systematic characterization of the giant panda URT virome, establishing a critical baseline for disease surveillance and highlighting the complex interplay between the host and its environment-associated viral elements.

Keywords

Giant panda / Upper respiratory tract / Virome / Phylogenetic analysis / Parvoviridae

Cite this article

Download citation ▾
Zidan Zeng, Hongwen Zhang, Shiyin Huang, Yunli Li, Yue Chen, Shixing Yang, Quan Shen, Xiaochun Wang, Yuwei Liu, Ping Wu, Likai Ji, Songrui Liu, Wen Zhang. Viral metagenomic analysis of the upper respiratory tract virome and identification of novel viruses in giant pandas. Animal Diseases, 2026, 6(1): 20 DOI:10.1186/s44149-026-00229-7

登录浏览全文

4963

注册一个新账户 忘记密码

References

[1]

Agius, J.E., Phalen, D.N., Rose, K., Eden, J.S., 2019. New insights into sauropsid papillomaviridae evolution and epizootiology: discovery of two novel papillomaviruses in native and invasive island geckos. Virus Evol. 5, vez051. https://doi.org/10.1093/ve/vez051

[2]

Barzon L, Lavezzo E, Militello V, Toppo S, Palù G. Applications of next-generation sequencing technologies to diagnostic virology. International Journal of Molecular Sciences, 2011, 12: 7861-7884

[3]

Buchfink B, Xie C, Huson DH. Fast and sensitive protein alignment using DIAMOND. Nature Methods, 2015, 12: 59-60

[4]

Canuti, M., Pénzes, J.J., Lang, A.S., 2022. A new perspective on the evolution and diversity of the genus amdoparvovirus (family parvoviridae) through genetic characterization, structural homology modeling, and phylogenetics. Virus Evol. 8, veac056. https://doi.org/10.1093/ve/veac056

[5]

Carlson, J., Suchman, E., Buchatsky, L., 2006. Densoviruses for control and genetic manipulation of mosquitoes, in: Advances in Virus Research, Insect Viruses: Biotechnological Applications. Academic Press, pp. 361–392. https://doi.org/10.1016/S0065-3527(06)68010-X

[6]

Chen Y, Huang S, Lu X, Ji L, Shen Q, Yang S, Liu Y, Wang X, Wu P, Yang H, Shan T, Zhang W. Viral metagenomics reveals parvovirus dark matter of herbivorous wildlife from the Qinghai-Tibet Plateau. BMC Microbiology, 2025, 25 508

[7]

Conceição-Neto N, Zeller M, Lefrère H, De Bruyn P, Beller L, Deboutte W, Yinda CK, Lavigne R, Maes P, et al. . Modular approach to customize sample preparation procedures for viral metagenomics: A reproducible protocol for virome analysis. Scientific Reports, 2015, 5: 1-14

[8]

Couto, R. da S., Abreu, W.U., Rodrigues, L.R.R., Marinho, L.F., Morais, V. dos S., Villanova, F., Pandey, R.P., Deng, X., et al. 2024. Genomoviruses in liver samples of molossus molossus bats. Microorganisms 12, 688. https://doi.org/10.3390/microorganisms12040688

[9]

Dai, Z., Wang, H., Wu, H., Zhang, Q., Ji, L., Wang, X., Shen, Q., Yang, S., Ma, X., Shan, T., Zhang, W., 2023. Parvovirus dark matter in the cloaca of wild birds. Giga Science 12, giad001. https://doi.org/10.1093/gigascience/giad001

[10]

Dai, Z., Wang, H., Xu, J., Lu, X., Ni, P., Yang, S., Shen, Q., Wang, Xiaochun, Li, W., et al. 2024. Unveiling the virome of wild birds: exploring CRESS-DNA viral dark matter. Genome Biol. Evol. 16, evae206. https://doi.org/10.1093/gbe/evae206

[11]

De Souza WM, Dennis T, Fumagalli MJ, Araujo J, Sabino-Santos G, Maia FGM, Acrani GO, Carrasco ADOT, Romeiro MF, Modha S, et al. . Novel parvoviruses from wild and domestic animals in Brazil provide new insights into parvovirus distribution and diversity. Viruses, 2018, 10: 143

[12]

Deng X, Naccache SN, Ng T, Federman S, Li L, Chiu CY, Delwart EL. An ensemble strategy that significantly improves de novo assembly of microbial genomes from metagenomic next-generation sequencing data. Nucleic Acids Research, 2015, 43 e46

[13]

Feng, Y., Gou, Q., Yang, W., Wu, W., Wang, J., Holmes, E.C., Liang, G., Shi, M., 2022. A time-series meta-transcriptomic analysis reveals the seasonal, host, and gender structure of mosquito viromes. Virus Evol. 8, veac006. https://doi.org/10.1093/ve/veac006

[14]

François S, Filloux D, Roumagnac P, Bigot D, Gayral P, Martin DP, Froissart R, Ogliastro M. Discovery of parvovirus-related sequences in an unexpected broad range of animals. Scientific Reports, 2016, 6 30880

[15]

Gao FS, Hu GX, Xia X, Gao YW, Bai YD, Zou XH. Isolation and identification of a canine coronavirus strain from giant pandas (ailuropoda melanoleuca). Journal of Veterinary Science, 2009, 10: 261-263

[16]

Hopkins M, Kailasan S, Cohen A, Roux S, Tucker KP, Shevenell A, Agbandje-McKenna M, Breitbart M. Diversity of environmental single-stranded DNA phages revealed by PCR amplification of the partial major capsid protein. ISME Journal, 2014, 8: 2093-2103

[17]

Huson DH, Auch AF, Qi J, Schuster SC. MEGAN analysis of metagenomic data. Genome Research, 2007, 17: 377-386

[18]

Khalil, A.M., Martinez-Sobrido, L., Mostafa, A., 2024. Zoonosis and zooanthroponosis of emerging respiratory viruses. Front. Cell. Infect. Microbiol. 13. https://doi.org/10.3389/fcimb.2023.1232772

[19]

Knezevic P, Adriaenssens EMICTV Report Consortium. ICTV virus taxonomy profile: Inoviridae. Journal of General Virology, 2021, 102 1614

[20]

Kumar S, Stecher G, Li M, Knyaz C, Tamura K. MEGA X: Molecular evolutionary genetics analysis across computing platforms. Molecular Biology and Evolution, 2018, 35: 1547-1549

[21]

Lan T, Yang S, Li H, Zhang Y, Li R, Sahu SK, Deng W, Liu B, Shi M, Wang S, et al. . Large-scale genome sequencing of giant pandas improves the understanding of population structure and future conservation initiatives. Proceedings of the National Academy of Sciences, 2024, 121 e2406343121

[22]

Langmead B, Salzberg SL. Fast gapped-read alignment with bowtie 2. Nature Methods, 2012, 9: 357-359

[23]

Letunic I, Bork P. Interactive tree of life (iTOL) v5: An online tool for phylogenetic tree display and annotation. Nucleic Acids Research, 2021, 49: W293-W296

[24]

Li D, Zhu L, Cui H, Ling S, Fan S, Yu Z, Zhou Y, Wang T, Qian J, Xia X, et al. . Influenza A(H1N1)pdm09 virus infection in giant pandas, China. Emerging Infectious Diseases, 2014, 20: 480-483

[25]

Li D, Liu C-M, Luo R, Sadakane K, Lam T-W. MEGAHIT: An ultrafast single-node solution for large and complex metagenomics assembly via succinct de bruijn graph. Bioinformatics, 2015, 31: 1674-1676

[26]

Li X, Chen M, Chen T, Xie L, Luo Q, Fan X, Yin Y, Meng S, Jin Z, He Y, Wen Y. The intricate interplay among microbiota, mucosal immunity, and viral infection in the respiratory tract. Journal of Translational Medicine, 2025, 23 488

[27]

Liu A, Qiao W, Ma R, Yan Q, Zhao S, Lang Y. The detection of mixed infection with canine parvovirus, canine distemper virus, and rotavirus in giant pandas by multiplex PCR. Veterinary Sciences, 2025, 12 81

[28]

Ma B, Lei S, Qing Q, Wen Y. Should the endangered status of the giant panda truly be reduced? The case of giant panda conservation in Sichuan, China. Animals, 2018, 8 69

[29]

Marchler-Bauer A, Derbyshire MK, Gonzales NR, Lu S, Chitsaz F, Geer LY, Geer RC, He J, Gwadz M, Hurwitz DI, et al. . CDD: NCBI’s conserved domain database. Nucleic Acids Research, 2015, 43: D222-D226

[30]

Ning S, Lu X, Zhao M, Wang X, Yang S, Shen Q, Wang H, Zhang W. Virome in fecal samples from wild giant pandas (ailuropoda melanoleuca). Frontiers in Veterinary Science, 2021, 8 767494

[31]

Pénzes JJ, de Souza WM, Agbandje-McKenna M, Gifford RJ. An ancient lineage of highly divergent parvoviruses infects both vertebrate and invertebrate hosts. Viruses, 2019, 11 525

[32]

Pénzes JJ, Söderlund-Venermo M, Canuti M, Eis-Hübinger AM, Hughes J, Cotmore SF, Harrach B. Reorganizing the family parvoviridae: A revised taxonomy independent of the canonical approach based on host association. Archives of Virology, 2020, 165: 2133-2146

[33]

Popgeorgiev N, Krupovic M, Hiblot J, Fancello L, Monteil-Bouchard S, Desnues C. A new inovirus from the human blood encodes proteins with nuclear subcellular localization. Viruses, 2024, 16 475

[34]

Popov Ilia V., Manakhov AD, Gorobets VE, Diakova KB, Lukbanova EA, Malinovkin AV, Venema K, Ermakov AM, Popov Igor V.. Metagenomic investigation of intestinal microbiota of insectivorous synanthropic bats: Densoviruses, antibiotic resistance genes, and functional profiling of gut microbial communities. International Journal of Molecular Sciences, 2025, 26 5941

[35]

Prado, T., Magalhães, M.G.P., Moreira, D.A., Brandão, M.L., Fumian, T.M., Ferreira, F.C., Chame, M., Leomil, L., Degrave, W.M.S., Leite, et al. 2025. Microbiome and virome on indoor surfaces of an antarctic research ship. Mem. Inst. Oswaldo Cruz 118, e230084. https://doi.org/10.1590/0074-02760230084

[36]

Quaiser, A., Dufresne, A., Ballaud, F., Roux, S., Zivanovic, Y., Colombet, J., Sime Ngando, T., Francez, A.J., 2015. Diversity and comparative genomics of microviridae in sphagnum- dominated peatlands. Front. Microbiol. 6. https://doi.org/10.3389/fmicb.2015.00375

[37]

Que, T., Pang, X., Huang, H., Chen, P., Wei, Y., Hua, Y., Liao, H., Wu, J., Li, S., Wu, A., He, M., Ruan, X., Hu, Y., 2022. Comparative gut microbiome in trachypithecus leucocephalus and other primates in guangxi, China, based on metagenome sequencing. Front. Cell. Infect. Microbiol. 12. https://doi.org/10.3389/fcimb.2022.872841

[38]

Rojas CA, Holekamp KE, Viladomat Jasso M, Souza V, Eisen JA, Theis KR. Taxonomic, genomic, and functional variation in the gut microbiomes of wild spotted hyenas across 2 decades of study. mSystems, 2022, 8 e00965-22

[39]

Ronquist F, Teslenko M, van der Mark P, Ayres DL, Darling A, Höhna S, Larget B, Liu L, Suchard MA, Huelsenbeck JP. MrBayes 3.2: Efficient bayesian phylogenetic inference and model choice across a large model space. Systematic Biology, 2012, 61: 539-542

[40]

Roux S, Krupovic M, Poulet A, Debroas D, Enault F. Evolution and diversity of the microviridae viral family through a collection of 81 new complete genomes assembled from virome reads. PLoS ONE, 2012, 7 e40418

[41]

Roux S, Krupovic M, Daly RA, Borges AL, Nayfach S, Schulz F, Sharrar A, Matheus Carnevali PB, Cheng JF, Ivanova NN, et al. . Cryptic inoviruses revealed as pervasive in bacteria and archaea across Earth’s biomes. Nature Microbiology, 2019, 4: 1895-1906

[42]

Sauthier JT, Daudt C, da Silva FRC, Alves CDBT, Mayer FQ, Bianchi RM, Driemeier D, Streit RSA, Staats CC, Canal CW, et al. . The genetic diversity of “papillomavirome” in bovine teat papilloma lesions. Anim. Microbiome, 2021, 3: 51

[43]

Schmieder R, Edwards R. Quality control and preprocessing of metagenomic datasets. Bioinformatics, 2011, 27: 863-864

[44]

Souza WMde, Romeiro MF, Fumagalli MJ, Modha S, de Araujo J, Queiroz LH, Durigon EL, Figueiredo LTM, Murcia PR, Gifford RJ. Chapparvoviruses occur in at least three vertebrate classes and have a broad biogeographic distribution. Journal of General Virology, 2017, 98: 225-229

[45]

Tominaga, K., Ogawa-Haruki, N., Nishimura, Y., Watai, H., Yamamoto, K., Ogata, H., Yoshida, T., n.d. Prevalence of viral frequency-dependent infection in coastal marine prokaryotes revealed using monthly time series virome analysis. Msystems 8, e00931–22. https://doi.org/10.1128/msystems.00931-22

[46]

Tucker KP, Parsons R, Symonds EM, Breitbart M. Diversity and distribution of single-stranded DNA phages in the north Atlantic Ocean. ISME Journal, 2011, 5: 822-830

[47]

Varsani A, Krupovic M. Family genomoviridae: 2021 taxonomy update. Archives of Virology, 2021, 166: 2911-2926

[48]

Wang, H., Ling, Y., Shan, T., Yang, S., Xu, H., Deng, X., Delwart, E., Zhang, W., 2019. Gut virome of mammals and birds reveals high genetic diversity of the family microviridae. Virus Evol. 5, vez013. https://doi.org/10.1093/ve/vez013

[49]

Yang Y, Geng Y, Ouyang P, Li Y, Guo H, Deng H, Hou R, Lai W, Zhang D, Liu S. Identification of a feline panleukopenia virus from captive giant pandas (ailuropoda melanoleuca) and its phylogenetic analysis. Transboundary Emerging Dis., 2023, 2023: 7721487

[50]

Zeng J, Wang Y, Zhang J, Yang S, Zhang W. Multiple novel filamentous phages detected in the cloacal swab samples of birds using viral metagenomics approach. Virology Journal, 2021, 18 240

[51]

Zhang W, Yang S, Shan T, Hou R, Liu Z, Li W, Guo L, Wang Y, Chen P, Wang X, et al. . Virome comparisons in wild-diseased and healthy captive giant pandas. Microbiome, 2017, 5 90

[52]

Zhang W, Liu W, Hou R, Zhang L, Schmitz-Esser S, Sun H, Xie J, Zhang Y, Wang C, Li L, Yue B, et al. . Age-associated microbiome shows the giant panda lives on hemicelluloses, not on cellulose. ISME Journal, 2018, 12: 1319-1328

[53]

Zhang L, Li C, Zhai Y, Feng L, Bai K, Zhang Z, Huang Y, Li T, Li D, Li H, Cui P, Chen D, Wang H, Yang X. Analysis of the vaginal microbiome of giant pandas using metagenomics sequencing. MicrobiologyOpen, 2020, 9 e1131

[54]

Zhao N, Li M, Luo J, Wang Supen, Liu S, Wang Shan, Lyu W, Chen L, Su W, Ding H, He H. Impacts of canine distemper virus infection on the giant panda population from the perspective of gut microbiota. Scientific Reports, 2017, 7 39954

[55]

Zhao M, Yue C, Yang Z, Li Y, Zhang D, Zhang J, Yang S, Shen Q, Su X, Qi D, et al. . Viral metagenomics unveiled extensive communications of viruses within giant pandas and their associated organisms in the same ecosystem. Science of the Total Environment, 2022, 820 153317

[56]

Zhao, S., Hu, H., Lan, J., Yang, Z., Peng, Q., Yan, L., Luo, L., Wu, L., Lang, Y., Yan, Q., 2023. Characterization of a fatal feline panleukopenia virus derived from giant panda with broad cell tropism and zoonotic potential. Front. Immunol. 14. https://doi.org/10.3389/fimmu.2023.1237630

[57]

Zhou Z, Shi X, Li K, Hu Q, Ren Y, Zhou X, Li M, Zhang T, Yang F, Huang Y, et al. . First identification of CPV-2c infection in a wild cub giant panda (ailuropoda melanoleuca) suggesting an emerging transmission from wildlife and domestic dogs. Transboundary and Emerging Diseases, 2025, 2025 6716483

Funding

Chongqing Municipal Key Research and Development Program of China(2023YFD1801300)

National Natural Science Foundation of China(82550118)

Chengdu Research Base of Giant Panda Breeding(202503KY0001)

RIGHTS & PERMISSIONS

The Author(s)

PDF

0

Accesses

0

Citation

Detail

Sections
Recommended

/