A metagenomic viral discovery approach identifies potential zoonotic and novel mammalian viruses in Neoromicia bats within South Africa

PLoS ONE

Volumn 13, Issue 3, 2018, Pages

  Geldenhuys, Marike ^a   Mortlock, Marinda ^a   Weyer, Jacqueline ^b   Bezuidt, Oliver ^a   Seamark, Ernest C J ^a,c   Kearney, Teresa ^d,e   Gleasner, Cheryl ^f   Erkkila, Tracy H ^f   Cui, Helen ^f   Markotter, Wanda ^a  

^a UNIVERSITY OF PRETORIA   (South Africa)

^b NATIONAL INSTITUTE FOR COMMUNICABLE DISEASES   (South Africa)

^c AfricanBats NPC   (South Africa)

^d DITSONG NATIONAL MUSEUM OF NATURAL HISTORY   (South Africa)

^e UNIVERSITY OF THE WITWATERSRAND   (South Africa)

^f LOS ALAMOS NATIONAL LABORATORY   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ADENOVIRIDAE; ANIMAL; BAT; BIOLOGY; CORONAVIRINAE; CORONAVIRUS INFECTION; DNA SEQUENCE; GASTROINTESTINAL TRACT; GENETIC VARIATION; GENETICS; HERPESVIRIDAE; HIGH THROUGHPUT SEQUENCING; HUMAN; MAMMAL; METAGENOMICS; PATHOGENICITY; PHYLOGENY; PHYLOGEOGRAPHY; PHYSIOLOGY; PROCEDURES; SOUTH AFRICA; VETERINARY; VIROLOGY; VIRUS GENOME; ZOONOSIS;

ADENOVIRIDAE; ANIMALS; CHIROPTERA; COMPUTATIONAL BIOLOGY; CORONAVIRUS; CORONAVIRUS INFECTIONS; GASTROINTESTINAL TRACT; GENETIC VARIATION; GENOME, VIRAL; HERPESVIRIDAE; HIGH-THROUGHPUT NUCLEOTIDE SEQUENCING; HUMANS; MAMMALS; METAGENOMICS; PHYLOGENY; PHYLOGEOGRAPHY; SEQUENCE ANALYSIS, DNA; SOUTH AFRICA; ZOONOSES;

EID: 85044544238     PISSN: None     EISSN: 19326203     Source Type: Journal    
DOI: 10.1371/journal.pone.0194527     Document Type: Article

References (82)

1. Rahman MA, Khan SU, Rahman M, Gurley ES, Rollin PE, Lo MK, et al. Date Palm Sap Linked to Nipah Virus Outbreak in Bangladesh, 2008. Vector borne zoonotic Dis. 2012; 12(1):65-72. https://doi.org/10.1089/vbz.2011.0656 PMID: 21923274
2. Amman BR, Carroll SA, Reed ZD, Sealy TK, Balinandi S, Swanepoel R, et al. Seasonal pulses of Marburg virus circulation in juvenile Rousettus aegyptiacus bats coincide with periods of increased risk of human infection. PLoS Pathog. 2012; 8(10).
3. Lau SKP, Woo PCY, Li KSM, Huang Y, Tsoi H-W, Wong BHL, et al. Severe acute respiratory syndrome coronavirus-like virus in Chinese horseshoe bats. PNAS. 2005; 102(39):14040-5. https://doi.org/10.1073/pnas.0506735102 PMID: 16169905
4. Corman VM, Ithete NL, Richards R, Schoeman MC, Preiser W, Drosten C, et al. Rooting the phylogenetic tree of Middle East respiratory syndrome coronavirus by characterization of a conspecific virus from an African bat. J Virol. 2014; 88(19):11297-303. https://doi.org/10.1128/JVI.01498-14 PMID: 25031349
5. Anthony SJ, Gilardi K, Menachery VD, Goldstein T, Ssebide B, Mbabazi R, et al. Further evidence for bats as the evolutionary source of Middle East respiratory syndrome coronavirus. MBio. 2017; 8(2):1-13.
6. Drexler JF, Corman VM, Drosten C. Ecology, evolution and classification of bat coronaviruses in the aftermath of SARS. Antiviral Res. 2014; 101(1):45-56.
7. Ge X, Li J, Yang X, Chmura AA, Zhu G, Epstein JH, et al. Isolation and Characterization of a bat SARSlike coronavirus that uses the ACE2 receptor. Nature. 2013; 503(7477):535-8. https://doi.org/10.1038/nature12711 PMID: 24172901
8. Wu Z, Yang L, Ren X, He G, Zhang J, Yang J, et al. Deciphering the bat virome catalog to better understand the ecological diversity of bat viruses and the bat origin of emerging infectious diseases. ISME J. 2015; 10:1-12.
9. Wang J, Moore NE, Murray ZL, Mcinnes K, White J, Tompkins DM, et al. Discovery of novel virus sequences in an isolated and threatened bat species, the New Zealand lesser short-tailed bat (Mystacina tuberculata). J Virol. 2015; 96:2442-52.
10. Drexler JF, Corman VM, Muèller MA, Maganga GD, Vallo P, Binger T, et al. Bats host major mammalian paramyxoviruses. Nat Commun. 2012; 3(796):1-12.
11. Wu Z, Ren X, Yang L, Hu Y, Yang J, He G, et al. Virome analysis for identification of novel mammalian viruses in bat species from Chinese provinces. J Virol. 2012; 86(20):10999-1012. https://doi.org/10.1128/JVI.01394-12 PMID: 22855479
12. Drexler JF, Geipel A, Koènig A, Corman VM, van Riel D, Leijten LM, et al. Bats carry pathogenic hepadnaviruses antigenically related to hepatitis B virus and capable of infecting human hepatocytes. PNAS. 2013; 110(40):16151-6. https://doi.org/10.1073/pnas.1308049110 PMID: 24043818
13. Tang XC, Zhang JX, Zhang SY, Wang P, Fan X. H, Li LF, et al. Prevalence and genetic diversity of coronaviruses in bats from China. J Virol. 2006; 80(15):7481-90. https://doi.org/10.1128/JVI.00697-06 PMID: 16840328
14. Li L, Victoria JG, Wang C, Jones M, Fellers GM, Kunz TH, et al. Bat guano virome: predominance of dietary viruses from insects and plants plus novel mammalian viruses. J Virol. 2010; 84(14):6955-65. https://doi.org/10.1128/JVI.00501-10 PMID: 20463061
15. Ge X, Li Y, Yang X, Zhang H, Zhou P, Zhang Y, et al. Metagenomic analysis of viruses from bat fecal samples reveals many novel viruses in insectivorous bats in China. J Virol. 2012; 86(8):4620-30. https://doi.org/10.1128/JVI.06671-11 PMID: 22345464
16. Donaldson EF, Haskew AN, Gates JE, Huynh J, Moore CJ, Frieman MB. Metagenomic analysis of the viromes of three North American bat species: viral diversity among different bat species that share a common habitat. J Virol. 2010; 84(24):13004-18. https://doi.org/10.1128/JVI.01255-10 PMID: 20926577
17. Baker KS, Leggett RM, Bexfield NH, Alston M, Daly G, Todd S, et al. Metagenomic study of the viruses of African straw-coloured fruit bats: Detection of a chiropteran poxvirus and isolation of a novel adenovirus. Virology. 2013; 441(2):95-106. https://doi.org/10.1016/j.virol.2013.03.014 PMID: 23562481
18. Dacheux L, Cervantes-Gonzalez M, Guigon G, Thiberge JM, Vandenbogaert M, Maufrais C, et al. A preliminary study of viral metagenomics of french bat species in contact with humans: Identification of new mammalian viruses. PLoS One. 2014; 9(1):e87194. https://doi.org/10.1371/journal.pone.0087194 PMID: 24489870
19. Kohl C, Vidovszky MZ, Muhldorfer K, Dabrowski PW, Radonić A, Nitsche A, et al. Genome analysis of bat adenovirus 2: indications of interspecies transmission. J Virol. 2012; 86(3):1888-92. https://doi.org/10.1128/JVI.05974-11 PMID: 22130531
20. Quan P, Firth C, Conte JM, Williams SH, Zambrana-Torrelio CM, Anthony SJ, et al. Bats are a major natural reservoir for hepaciviruses and pegiviruses. PNAS. 2013; 110(20):8194-9. https://doi.org/10.1073/pnas.1303037110 PMID: 23610427
21. ACR. African Chiroptera report. Van Cakenberghe V, Seamark ECJ, editors. Pretoria: AfricanBats, Pretoria, Republic of South Africa ISSN: 1990-6471; 2016.
22. Goodman SM, Kearney T, Ratsimbazafy MM, Hassanin A. Description of a new species of Neoromicia (Chiroptera: Vespertilionidae) from southern Africa: A name for N. cf. melckorum. Zootaxa. 2017; 4236 (2):zootaxa.4236.2.10.
23. Jacobs D, Cotterill FPD, Taylor PJ. Neoromicia capensis. The IUCN Red List of Threatened Species 2014: e.T44918A67358046. 2014.
24. Hutson AM, Racey PA, Goodman S, Jacobs D. Neoromicia nana. The IUCN Red List of Threatened Species 2014: e.T44923A67357605. 2014.
25. Sumibcay L, Kadjo B, Gu SH, Kang HJ, Lim BK, Cook JA, et al. Divergent lineage of a novel hantavirus in the banana pipistrelle (Neoromicia nanus) in Cote d'Ivoire. Virol J. 2012; 9(34).
26. Mortlock M, Kuzmin I V, Weyer J, Gilbert AT, Agwanda B, Rupprecht CE, et al. Novel paramyxoviruses in bats from sub-Saharan Africa, 2007-2012. Emerg Infect Dis. 2015; 21(10):1840-3. https://doi.org/10.3201/eid2110.140368 PMID: 26402433
27. Geldenhuys M, Weyer J, Nel LH, Markotter W. Coronaviruses in South African bats. Vector borne zoonotic Dis. 2013; 13(7):516-9. https://doi.org/10.1089/vbz.2012.1101 PMID: 23473214
28. Ithete NL, Stoffberg S, Corman VM, Cottontail VM, Richards LR, Schoeman MC, et al. Close relative of human Middle East respiratory syndrome coronavirus in bat, South Africa. Emerg Infect Dis. 2013; 19 (10):1697-9. https://doi.org/10.3201/eid1910.130946 PMID: 24050621
29. Gloza-Rausch F, Ipsen A, Seebens A, Goèttsche M, Panning M, Felix Drexler J, et al. Detection and prevalence patterns of group I coronaviruses in bats, northern Germany. Emerg Infect Dis. 2008; 14 (4):626-31. https://doi.org/10.3201/eid1404.071439 PMID: 18400147
30. Monadjem A, Taylor PJ, Cotterill W, Schoeman MC. Bats of southern and central Africa: a biogeographic and taxonomic synthesis. Johannesburg: Wits University Press; 2010.
31. Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek R. DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Mol Mar Biol Biotechnol. 1994; 3 (5):294-9. PMID: 7881515
32. Kohl C, Brinkmann A, Dabrowski PW, Radoni A, Nitsche A, Kurth A. Protocol for metagenomic virus detection in clinical specimens. Emerg Infect Dis. 2015; 21(1):49-57.
33. Victoria JG, Kapoor A, Dupuis K, Schnurr DP, Delwart EL. Rapid identification of known and new RNA viruses from animal tissues. PLoS Pathog. 2008; 4(9):e1000163. https://doi.org/10.1371/journal.ppat. 1000163 PMID: 18818738
34. Huson DH, Auch AF, Qi J, Schuster SC. MEGAN analysis of metagenomic data. 2007;377-86.
35. Zerbino DR, Birney E. Velvet: Algorithms for de novo short read assembly using de Bruijn graphs. Genome Res. 2008; 18(5):821-9. https://doi.org/10.1101/gr.074492.107 PMID: 18349386
36. Drexler JF, Gloza-Rausch F, Glende J, Corman VM, Muth D, Goettsche M, et al. Genomic characterization of severe acute respiratory syndrome-related coronavirus in European bats and classification of coronaviruses based on partial RNA-dependent RNA polymerase gene sequences. J Virol. 2010; 84(21):11336-49. https://doi.org/10.1128/JVI.00650-10 PMID: 20686038
37. Li Y, Ge X, Zhang H, Zhou P, Zhu Y, Zhang Y, et al. Host range, prevalence, and genetic diversity of adenoviruses in bats. J Virol. 2010; 84(8):3889-97. https://doi.org/10.1128/JVI.02497-09 PMID: 20089640
38. Van Devanter DR, Warrener P, Bennett L, Schultz ER, Coulter S, Garber RL, et al. Detection and analysis of diverse herpesviral species by consensus primer PCR. J Clin Microbiol. 1996; 34(7):1666-71. PMID: 8784566
39. Wang S, Sundaram JP, Spiro D. VIGOR, an annotation program for small viral genomes. BMC Bioinformatics. 2010; 11(451):1-10.
40. Hall T. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser. 1999; 41:95-8.
41. Kumar S, Stecher G, Tamura K. MEGA7: Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Mol Biol Evol. 2016; 33(7):1870-4. https://doi.org/10.1093/molbev/msw054 PMID: 27004904
42. Drummond AJ, Suchard MA, Xie D, Rambaut A. Bayesian Phylogenetics with BEAUti and the BEAST 1.7. Mol Biol Evol. 2012; 29(8):1969-73. https://doi.org/10.1093/molbev/mss075 PMID: 22367748
43. Darriba D, Taboada GL, Doallo R, Posada D. jModelTest 2: more models, new heuristics and high-performance computing. Nat Methods. 2015; 9(8):6-9.
44. Miller MA, Pfeiffer W, Schwartz T. Creating the CIPRES science gateway for inference of large phylogenetic trees. In: Proceedings of the Gateway Computing Environments Workshop (GCE). New Orleans, LA; 2010. p. 1-8.
45. Sayre R, Comer P, Hak J, Josse C, Bow J, Warner H, et al. A new map of standardized terrestrial ecosystems of Africa. African Geogr Rev. 2013;1-24.
46. Faye B. Role, distribution and perspective of camel breeding in the third millennium economies. Emirates J Food Agric. 2015; 27(4):318-27.
47. Wilson RT. The one-humped camel in Southern Africa: Unusual and new records for seven countries in the Southern African development community. African J Agric Res. 2013; 8(28):3716-23.
48. Hijmans RJ, Cameron SE, Parra JL, Jones PG, Jarvis A. Very high resolution interpolated climate surfaces for global land areas. Int J Climatol. 2005; 25:1965-78.
49. Phillips SB, Aneja VP, Kang D, Arya SP. Modelling and analysis of the atmospheric nitrogen deposition in North Carolina. Int J Glob Environ Issues. 2006; 6(2-3):231-52.
50. Reusken CBEM, Haagmans BL, Muèller MA, Gutierrez C, Godeke G-J, Meyer B, et al. Middle East respiratory syndrome coronavirus in dromedary camels: an outbreak investigation. Lancet. 2013; 3099 (13):1-6.
51. Reusken CBEM, Messadi L, Feyisa A, Ularamu H, Godeke GJ, Danmarwa A, et al. Geographic distribution of MERS coronavirus among dromedary camels, Africa. Emerg Infect Dis. 2014; 20(8):1370-4. https://doi.org/10.3201/eid2008.140590 PMID: 25062254
52. Corman VM, Jores J, Meyer B, Younan M, Liljander A, Said MY, et al. Antibodies against MERS coronavirus in Dromedary camels, Kenya, 1992-2013. Emerg Infect Dis. 2014; 20(8):1319-22. https://doi.org/10.3201/eid2008.140596 PMID: 25075637
53. Muèller MA, Corman VM, Jores J, Meyer B, Younan M, Liljander A, et al. MERS coronavirus neutralizing antibodies in camels, eastern Africa, 1983-1997. Emerg Infect Dis. 2014; 20(12):2093-5. https://doi.org/10.3201/eid2012.141026 PMID: 25425139
54. Chu DKW, Poon LLM, Gomaa MM, Shehata MM, Perera RAPM, Zeid DA, et al. MERS coronaviruses in dromedary camels, Egypt. Emerg Infect Dis. 2014; 20(6):1049-53. https://doi.org/10.3201/eid2006. 140299 PMID: 24856660
55. Hemida MG, Elmoslemany A, Al-Hizab F, Alnaeem A, Almathen F, Faye B, et al. Dromedary camels and the transmission of Middle East respiratory syndrome coronavirus (MERS-CoV). Transbound Emerg Dis. 2015; 64(2):344-53. https://doi.org/10.1111/tbed.12401 PMID: 26256102
56. He B, Li Z, Yang F, Zheng J, Feng Y, Guo H, et al. Virome profiling of bats from myanmar by metagenomic analysis of tissue samples reveals more novel Mammalian viruses. PLoS One. 2013; 8(4): e61950. https://doi.org/10.1371/journal.pone.0061950 PMID: 23630620
57. Zhao G, Wu G, Lim ES, Droit L, Krishnamurthy S, Barouch DH, et al. Virus Seeker, a computational pipeline for virus discovery and virome composition analysis. Virology. 2017; 503:21-30. https://doi.org/10.1016/j.virol.2017.01.005 PMID: 28110145
58. Li L, Kapoor A, Slikas B, Bamidele OS, Wang C, Shaukat S, et al. Multiple diverse circoviruses infect farm animals and are commonly found in human and chimpanzee feces. J Virol. 2010; 84(4):1674-82. https://doi.org/10.1128/JVI.02109-09 PMID: 20007276
59. de S Lima FE, Cibulski SP, dos Santos HF, Teixeira TF, Varela APM, Roehe PM, et al. Genomic characterization of novel circular ssDNA viruses from insectivorous bats in Southern Brazil. PLoS One. 2015; 10(2):e0118070. https://doi.org/10.1371/journal.pone.0118070 PMID: 25688970
60. Oude Munnink BB, Phan MVT, van der Hoek L, Kellam P, Cotten M. Genome Sequences of a Novel Vietnamese Bat Bunyavirus. Genome Announc. 2016; 4(6):e01366-16. https://doi.org/10.1128/genomeA.01366-16 PMID: 28007850
61. Mourya DT, Yadav PD, Basu A, Shete A, Patil DY, Zawar D, et al. Malsoor Virus, a novel bat phlebovirus, is closely related to Severe Fever with Thrombocytopenia Syndrome virus and Heartland virus. J Virol. 2014; 88(6):3605-9. https://doi.org/10.1128/JVI.02617-13 PMID: 24390329
62. Lau SKP, Woo PCY, Lai KKY, Huang Y, Yip CCY, Shek C-T, et al. Complete genome analysis of three novel picornaviruses from diverse bat species. J Virol. 2011; 85(17):8819-28. https://doi.org/10.1128/JVI.02364-10 PMID: 21697464
63. Kemenesi G, Zhang D, Marton S, Dallos B, Gorfol T, Estok P, et al. Genetic characterization of a novel picornavirus detected in Miniopterus schreibersii bats. J Gen Virol. 2014; 96:815-21. https://doi.org/10.1099/jgv.0.000028 PMID: 25516541
64. Lukashev AN, Corman VM, Schacht D, Gloza-rausch F, Seebens-hoyer A, Gmyl AP, et al. Close genetic relatedness of picornaviruses from European and Asian bats. J Gen Virol. 2017; 98:955-61. https://doi.org/10.1099/jgv.0.000760 PMID: 28555547
65. Pankovics P, Boros AÂ, Bõâroâ H, Barbara K, Gia T, Delwart E, et al. Novel picornavirus in domestic rabbits (Oryctolagus cuniculus var. domestica). Infect Genet Evol. 2016; 37:117-22. https://doi.org/10.1016/j. meegid.2015.11.012 PMID: 26588888
66. Li L, Deng X, Mee ET, Collot-teixeira S, Anderson R, Schepelmann S, et al. Comparing viral metagenomics methods using a highly multiplexed human viral pathogens reagent. J Virol Methods. 2015; 213:139-46. https://doi.org/10.1016/j.jviromet.2014.12.002 PMID: 25497414
67. Grard G, Fair JN, Lee D, Slikas E, Steffen I, Muyembe JJ, et al. A novel rhabdovirus sssociated with scute hemorrhagic fever in Central Africa. PLoS Pathog. 2012; 8(9):e1002924. https://doi.org/10.1371/journal.ppat.1002924 PMID: 23028323
68. Yuan L, Li M, Li L, Monagin C, Chmura AA, Schneider BS, et al. Evidence for retrovirus and paramyxovirus infection of multiple bat species in China. Viruses. 2014; 6:2138-54. https://doi.org/10.3390/v6052138 PMID: 24841387
69. Jaânoska M, Vidovszky M, Molnaâr V, Liptovszky M, Harrach B, Benko M. Novel adenoviruses and herpesviruses detected in bats. Vet J. 2011; 189(1):118-21. https://doi.org/10.1016/j.tvjl.2010.06.020 PMID: 20813566
70. Vidovszky MZ, Kohl C, Bologh S, Gorfol T, Wibbelt G, Kurth A, et al. Random sampling of the Central European bat fauna reveals the existence of numerous hithero unknown adenoviruses. Acta Vet Hung. 2015; 63(4):508-25. https://doi.org/10.1556/004.2015.047 PMID: 26599097
71. Benko M, Harrach B, Kremer EJ. Do nonhuman primate or bat adenoviruses pose a risk for human health? Future Microbiol. 2014; 9:269-72. https://doi.org/10.2217/fmb.13.170 PMID: 24762301
72. Sonntag M, Muèhldorfer K, Speck S, Wibbelt G, Kurth A. New adenovirus in bats, Germany. Emerg Infect Dis. 2009; 15(12):2052-5. https://doi.org/10.3201/eid1512.090646 PMID: 19961700
73. Shabman RS, Shrivastava S, Tsibane T, Attie O, Jayaprakash A, Mire CE, et al. Isolation and characterization of a novel Gammaherpesvirus from a microbat cell line. mSphere. 2016; 1(1):1-15.
74. Zheng X, Qiu M, Chen S, Xiao J, Ma L, Shan L, et al. High prevalence and diversity of viruses of the subfamily Gammaherpesvirinae, family Herpesviridae, in fecal specimens from bats of different species in southern China. Arch Virol. 2016; 161:135-40. https://doi.org/10.1007/s00705-015-2614-9 PMID: 26446885
75. Escalera-Zamudio M, Rojas-Anaya E, Kolokotronis S-O, Taboada B, Loza-Rubio E, Meândez-Ojeda ML, et al. Bats, primates, and the evolutionary origins and diversification of mammalian gammaherpesviruses. MBio. 2016; 7(6):e01425-16. https://doi.org/10.1128/mBio.01425-16 PMID: 27834200
76. de Groot RJ, Baker SC, Baric R, Enjuanes L, Gorbalenya AE, Holmes K V., et al. Family Coronaviridae. In: King A, Adams MJ, Carstens EB, Lefkowitz E, editors. Virus Taxonomy: Classification and nomenclature of viruses Ninth Report of the International Committee on Taxonomy of Viruses. London, United Kingdom: Academic Press; 2012. p. 806-20.
77. van der Hoek L, Pyrc K, Jebbink MF, Vermeulen-Oost W, Berkhout RJM, Wolthers KC, et al. Identification of a new human coronavirus. Nat Med. 2004; 10(4):368-73. https://doi.org/10.1038/nm1024 PMID: 15034574
78. Tao Y, Shi M, Chommanard C, Queen K, Zhang J, Markotter W, et al. Surveillance of bat coronaviruses in Kenya identifies relatives of human coronaviruses NL63 and 229E and their recombination history. J Virol. 2017; 91:e01953-16. https://doi.org/10.1128/JVI.01953-16 PMID: 28077633
79. Pfefferle S, Oppong S, Drexler JF, Gloza-Rausch F, Ipsen A, Seebens A, et al. Distant relatives of severe acute respiratory syndrome coronavirus and close relatives of human coronavirus 229E in bats, Ghana. Emerg Infect Dis. 2009; 15(9):1377-84. https://doi.org/10.3201/eid1509.090224 PMID: 19788804
80. Annan A, Baldwin HJ, Corman VM, Klose SM, Owusu M, Nkrumah EE, et al. Human Betacoronavirus 2c EMC/2012-related viruses in bats, Ghana and Europe. Emerg Infect Dis. 2013; 19(3):3-6.
81. Sabir JSM, Lam TT-Y, Ahmed MMM, Li L, Shen Y, E. M. Abo-Aba S, et al. Co-circulation of three camel coronavirus species and recombination of MERS-CoVs in Saudi Arabia. Science. 2015; 351(6268):81-4. https://doi.org/10.1126/science.aac8608 PMID: 26678874
82. Temmam S, Davoust B, Berenger J, Raoult D, Desnues C. Viral metagenomics on animals as a tool for the detection of zoonoses prior to human infection? Int J Mol Sci. 2014; 15:10377-97. https://doi.org/10.3390/ijms150610377 PMID: 24918293