Complete Genome Sequencing Reveals Unusual Equine Rotavirus A of Bat Origin from India.
Abstract: Rotaviruses are the most common viral agents associated with foal diarrhea. Between 2014 and 2017, the annual prevalence of rotavirus in diarrheic foals ranged between 18 and 28% in Haryana (India). Whole-genome sequencing of two equine rotavirus A (ERVA) isolates (RVA/Horse-wt/IND/ERV4/2017 and RVA/Horse-wt/IND/ERV6/2017) was carried out to determine the genotypic constellations (GCs) of ERVAs. The GCs of both the isolates were G3-P[3]-I8-R3-C3-M3-A9-N3-T3-E3-H6, a unique combination reported for ERVAs so far. Both the isolates carried VP6 of genotype I8, previously unreported from equines. Upon comparison with RVAs of other species, the GC of both isolates was identical to that of a bat rotavirus strain, MSLH14, isolated from China in 2012. The nucleotide sequences of the genes encoding VP3, NSP2, and NSP3 shared >95.81% identity with bat RVA strains isolated from Africa (Gabon). The genes encoding VP1, VP2, VP7, NSP1, and NSP4 shared 94.82% to 97.12% nucleotide identities with the human strains which have zoonotic links to bats (RCH272 and MS2015-1-0001). The VP6 genes of both strains were distinct and had the highest similarity of only 87.08% with that of CMH222, a human strain of bat origin. The phylogenetic analysis and lineage studies revealed that VP7 of both isolates clustered in a new lineage (lineage X) of the G3 genotype with bat, human, and alpaca strains. Similarly, VP4 clustered in a distinct P[3] lineage. These unusual findings highlight the terra incognita of the genomic diversity of equine rotaviruses and support the need for the surveillance of RVAs in animals and humans with a "one health" approach. Rotaviruses are globally prevalent diarrheal pathogens in young animals including foals, piglets, calves, goats, sheep, cats, and dogs along with humans. The genome of rotaviruses consists of 11 segments, which enables them to undergo reshuffling by reassortment of segments from multiple species during mixed infections. In this study, the prevalence of equine rotaviruses was 32.11% in organized equine farms of North India. The complete genome analysis of two ERVA isolates revealed an unusual genomic constellation, which was previously reported only in a bat RVA strain. A segment-wise phylogenetic analysis revealed that most segments of both isolates were highly similar either to bat or to bat-like human rotaviruses. The occurrence of unusual bat-like rotaviruses in equines emphasizes the need of extensive surveillance of complete genomes of both animal and human rotaviruses with a "one health" approach.
Publication Date: 2022-10-10 PubMed ID: 36214578PubMed Central: PMC9599578DOI: 10.1128/jvi.01408-22Google Scholar: Lookup
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Summary
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This research study conducted complete genetic sequencing on two isolates of equine rotavirus A (ERVA), a common cause of diarrhea in foals, in North India. The discovered genetic composition was unusually similar to a bat strain, marking the first time such similarity is reported in ERVA strains, highlighting the need for comprehensive examination considering the “one health” approach.
Background and Objectives
- The study arose from the need to examine the genetic structure or genotypic constellations (GCs) of ERVA, the leading cause of diarrhea in young horses, prevalent in North India.
- Rotaviruses have a complex genomic structure, facilitating genetic reshuffling, leading to the possibility of hybrid strains. Therefore, understanding ERVA strains’ genetic composition is crucial.
Methods
- Researchers performed a whole-genome sequencing of two ERVA isolates (RVA/Horse-wt/IND/ERV4/2017 and RVA/Horse-wt/IND/ERV6/2017).
- They then compared the genotypic constellations (GCs) of these isolates with other known rotavirus strains from various species.
Findings
- Both isolates exhibited a unique combination of GCs (G3-P[3]-I8-R3-C3-M3-A9-N3-T3-E3-H6) not previously reported in ERVAs.
- The nucleotide sequences for several encoding genes in the isolates showed high (>95.81%) identity with bat rotavirus stains.
- Certain gene sequences demonstrated a similarity of 94.82% to 97.12% with bat-derived human strains characterized by their zoonotic links (transmissible from animals to humans).
- The part of the genome responsible for the assembly of the viral shell (VP6) found in both strains was unique and only had a similarity of 87.08% with human strains originating from bats.
Implications
- The study unveils the unusual genetic diversity among equine rotaviruses, necessitating a more comprehensive approach to their surveillance.
- This discovery underlines the potential for viral transmission or reshuffling between species, particularly from bats to horses.
- The study emphasizes the importance of integrative surveillance of rotaviruses in animals and humans under the “one health” approach, considering the interconnectedness of human, animal, and environmental health.
Cite This Article
APA
Pathak A, Gulati BR, Maan S, Mor S, Kumar D, Soman R, Punia S, Chaudhary D, Khurana SK.
(2022).
Complete Genome Sequencing Reveals Unusual Equine Rotavirus A of Bat Origin from India.
J Virol, 96(20), e0140822.
https://doi.org/10.1128/jvi.01408-22 Publication
Researcher Affiliations
- Indian Council of Agricultural Research-National Research Centre on Equines, Hisar, Haryana, India.
- Indian Council of Agricultural Research-National Research Centre on Equines, Hisar, Haryana, India.
- Department of Animal Biotechnology, College of Veterinary Sciences, LLR University of Veterinary and Animal Sciences, Hisar, Haryana, India.
- Department of Veterinary Population Medicine, University of Minnesotagrid.17635.36, St. Paul, Minnesota, USA.
- Veterinary Diagnostic Laboratory, University of Minnesotagrid.17635.36, St. Paul, Minnesota, USA.
- Equine Breeding Stud, Hisar, Haryana, India.
- Indian Council of Agricultural Research-National Research Centre on Equines, Hisar, Haryana, India.
- Indian Council of Agricultural Research-National Research Centre on Equines, Hisar, Haryana, India.
- Department of Animal Biotechnology, College of Veterinary Sciences, LLR University of Veterinary and Animal Sciences, Hisar, Haryana, India.
- Indian Council of Agricultural Research-National Research Centre on Equines, Hisar, Haryana, India.
MeSH Terms
- Animals
- Horses / genetics
- Humans
- Sheep
- Swine
- Dogs
- Rotavirus
- Chiroptera / genetics
- Rotavirus Infections / epidemiology
- Rotavirus Infections / veterinary
- Rotavirus Infections / genetics
- Phylogeny
- Genome, Viral
- Diarrhea / veterinary
- Genotype
- Whole Genome Sequencing
- Camelids, New World / genetics
- Goats / genetics
- Nucleotides
- Gabon
Conflict of Interest Statement
The authors declare no conflict of interest.
References
This article includes 62 references
- Troeger C, Khalil IA, Rao PC, Cao S, Blacker BF, Ahmed T, Armah G, Bines JE, Brewer TG, Colombara DV, Kang G, Kirkpatrick BD, Kirkwood CD, Mwenda JM, Parashar UD, Petri WA Jr, Riddle MS, Steele AD, Thompson RL, Walson JL, Sanders JW, Mokdad AH, Murray CJL, Hay SI, Reiner RC Jr. Rotavirus Vaccination and the Global Burden of Rotavirus Diarrhea Among Children Younger Than 5 Years.. JAMA Pediatr 2018 Oct 1;172(10):958-965.
- Tate JE, Burton AH, Boschi-Pinto C, Parashar UD. Global, Regional, and National Estimates of Rotavirus Mortality in Children <5 Years of Age, 2000-2013.. Clin Infect Dis 2016 May 1;62 Suppl 2:S96-S105.
- John J, Sarkar R, Muliyil J, Bhandari N, Bhan MK, Kang G. Rotavirus gastroenteritis in India, 2011-2013: revised estimates of disease burden and potential impact of vaccines.. Vaccine 2014 Aug 11;32 Suppl 1:A5-9.
- Miño S, Adúriz M, Barrandeguy M, Parreño V. Molecular characterization of equine rotavirus group A detected in Argentinean foals during 2009–2014. J Equine Vet Sci 59:64–70.
- Desselberger U, Iturriza-Gómara M, Gray JJ. Rotavirus epidemiology and surveillance.. Novartis Found Symp 2001;238:125-47; discussion 147-52.
- Walker PJ, Siddell SG, Lefkowitz EJ, Mushegian AR, Dempsey DM, Dutilh BE, Harrach B, Harrison RL, Hendrickson RC, Junglen S, Knowles NJ, Kropinski AM, Krupovic M, Kuhn JH, Nibert M, Rubino L, Sabanadzovic S, Simmonds P, Varsani A, Zerbini FM, Davison AJ. Changes to virus taxonomy and the International Code of Virus Classification and Nomenclature ratified by the International Committee on Taxonomy of Viruses (2019).. Arch Virol 2019 Sep;164(9):2417-2429.
- Johne R, Tausch SH, Grützke J, Falkenhagen A, Patzina-Mehling C, Beer M, Höper D, Ulrich RG. Distantly Related Rotaviruses in Common Shrews, Germany, 2004-2014.. Emerg Infect Dis 2019 Dec;25(12):2310-2314.
- Johne R, Schilling-Loeffler K, Ulrich RG, Tausch SH. Whole Genome Sequence Analysis of a Prototype Strain of the Novel Putative Rotavirus Species L.. Viruses 2022 Feb 24;14(3).
- Bailey KE, Gilkerson JR, Browning GF. Equine rotaviruses--current understanding and continuing challenges.. Vet Microbiol 2013 Nov 29;167(1-2):135-44.
- Uprety T, Sreenivasan CC, Hause BM, Li G, Odemuyiwa SO, Locke S, Morgan J, Zeng L, Gilsenan WF, Slovis N, Metcalfe L, Carter CN, Timoney P, Horohov D, Wang D, Erol E, Adam E, Li F. Identification of a Ruminant Origin Group B Rotavirus Associated with Diarrhea Outbreaks in Foals.. Viruses 2021 Jul 9;13(7).
- Hoshino Y, Kapikian AZ. Classification of rotavirus VP4 and VP7 serotypes.. Arch Virol Suppl 1996;12:99-111.
- Hoshino Y, Wyatt RG, Greenberg HB, Flores J, Kapikian AZ. Serotypic similarity and diversity of rotaviruses of mammalian and avian origin as studied by plaque-reduction neutralization.. J Infect Dis 1984 May;149(5):694-702.
- Browning GF, Fitzgerald TA, Chalmers RM, Snodgrass DR. A novel group A rotavirus G serotype: serological and genomic characterization of equine isolate FI23.. J Clin Microbiol 1991 Sep;29(9):2043-6.
- Isa P, Wood AR, Netherwood T, Ciarlet M, Imagawa H, Snodgrass DR. Survey of equine rotaviruses shows conservation of one P genotype in background of two G genotypes.. Arch Virol 1996;141(9):1601-12.
- Ghosh S, Taniguchi K, Aida S, Ganesh B, Kobayashi N. Whole genomic analyses of equine group A rotaviruses from Japan: evidence for bovine-to-equine interspecies transmission and reassortment events.. Vet Microbiol 2013 Oct 25;166(3-4):474-85.
- Imagawa H, Ishida S, Uesugi S, Masanobu K, Fukunaga Y, Nakagomi O. Genetic analysis of equine rotavirus by RNA-RNA hybridization.. J Clin Microbiol 1994 Aug;32(8):2009-12.
- Hardy ME, Gorziglia M, Woode GN. The outer capsid protein VP4 of equine rotavirus strain H-2 represents a unique VP4 type by amino acid sequence analysis.. Virology 1993 Mar;193(1):492-7.
- Isa P, Snodgrass DR. Serological and genomic characterization of equine rotavirus VP4 proteins identifies three different P serotypes.. Virology 1994 Jun;201(2):364-72.
- Ghosh S, Shintani T, Kobayashi N. Evidence for the porcine origin of equine rotavirus strain H-1.. Vet Microbiol 2012 Aug 17;158(3-4):410-4.
- Nemoto M, Niwa H, Kida H, Higuchi T, Orita Y, Sato S, Bannai H, Tsujimura K, Ohta M. Isolation and characterization of a rare group A rotavirus G13P[18] strain from a diarrhoeic foal in Japan.. J Gen Virol 2020 Aug;101(8):800-805.
- Nemoto M, Tsunemitsu H, Imagawa H, Hata H, Higuchi T, Sato S, Orita Y, Sugita S, Bannai H, Tsujimura K, Yamanaka T, Kondo T, Matsumura T. Molecular characterization and analysis of equine rotavirus circulating in Japan from 2003 to 2008.. Vet Microbiol 2011 Aug 26;152(1-2):67-73.
- Matthijnssens J, Miño S, Papp H, Potgieter C, Novo L, Heylen E, Zeller M, Garaicoechea L, Badaracco A, Lengyel G, Kisfali P, Cullinane A, Collins PJ, Ciarlet M, O'Shea H, Parreño V, Bányai K, Barrandeguy M, Van Ranst M. Complete molecular genome analyses of equine rotavirus A strains from different continents reveal several novel genotypes and a largely conserved genotype constellation.. J Gen Virol 2012 Apr;93(Pt 4):866-875.
- Dóró R, Farkas SL, Martella V, Bányai K. Zoonotic transmission of rotavirus: surveillance and control.. Expert Rev Anti Infect Ther 2015;13(11):1337-50.
- Sadiq A, Bostan N, Yinda KC, Naseem S, Sattar S. Rotavirus: Genetics, pathogenesis and vaccine advances.. Rev Med Virol 2018 Nov;28(6):e2003.
- McDonald SM, Nelson MI, Turner PE, Patton JT. Reassortment in segmented RNA viruses: mechanisms and outcomes.. Nat Rev Microbiol 2016 Jul;14(7):448-60.
- Hoxie I, Dennehy JJ. Intragenic recombination influences rotavirus diversity and evolution.. Virus Evol 2020 Jan;6(1):vez059.
- Matthijnssens J, Ciarlet M, Heiman E, Arijs I, Delbeke T, McDonald SM, Palombo EA, Iturriza-Gómara M, Maes P, Patton JT, Rahman M, Van Ranst M. Full genome-based classification of rotaviruses reveals a common origin between human Wa-Like and porcine rotavirus strains and human DS-1-like and bovine rotavirus strains.. J Virol 2008 Apr;82(7):3204-19.
- Nemoto M, Nagai M, Tsunemitsu H, Omatsu T, Furuya T, Shirai J, Kondo T, Fujii Y, Todaka R, Katayama K, Mizutani T. Whole-genome sequence analysis of G3 and G14 equine group A rotaviruses isolated in the late 1990s and 2009-2010.. Arch Virol 2015 May;160(5):1171-9.
- Ma Y, Wen X, Hoshino Y, Yuan L. Cloning and nucleotide sequence analyses of 11 genome segments of two American and one British equine rotavirus strains.. Vet Microbiol 2015 Mar 23;176(1-2):172-8.
- Miño S, Matthijnssens J, Badaracco A, Garaicoechea L, Zeller M, Heylen E, Van Ranst M, Barrandeguy M, Parreño V. Equine G3P[3] rotavirus strain E3198 related to simian RRV and feline/canine-like rotaviruses based on complete genome analyses.. Vet Microbiol 2013 Jan 25;161(3-4):239-46.
- He B, Yang F, Yang W, Zhang Y, Feng Y, Zhou J, Xie J, Feng Y, Bao X, Guo H, Li Y, Xia L, Li N, Matthijnssens J, Zhang H, Tu C. Characterization of a novel G3P[3] rotavirus isolated from a lesser horseshoe bat: a distant relative of feline/canine rotaviruses.. J Virol 2013 Nov;87(22):12357-66.
- Xia L, Fan Q, He B, Xu L, Zhang F, Hu T, Wang Y, Li N, Qiu W, Zheng Y, Matthijnssens J, Tu C. The complete genome sequence of a G3P[10] Chinese bat rotavirus suggests multiple bat rotavirus inter-host species transmission events.. Infect Genet Evol 2014 Dec;28:1-4.
- Simsek C, Corman VM, Everling HU, Lukashev AN, Rasche A, Maganga GD, Binger T, Jansen D, Beller L, Deboutte W, Gloza-Rausch F, Seebens-Hoyer A, Yordanov S, Sylverken A, Oppong S, Sarkodie YA, Vallo P, Leroy EM, Bourgarel M, Yinda KC, Van Ranst M, Drosten C, Drexler JF, Matthijnssens J. At Least Seven Distinct Rotavirus Genotype Constellations in Bats with Evidence of Reassortment and Zoonotic Transmissions.. mBio 2021 Jan 19;12(1).
- Doan YH, Nakagomi T, Aboudy Y, Silberstein I, Behar-Novat E, Nakagomi O, Shulman LM. Identification by full-genome analysis of a bovine rotavirus transmitted directly to and causing diarrhea in a human child.. J Clin Microbiol 2013 Jan;51(1):182-9.
- Malik YS, Kumar N, Sharma K, Saurabh S, Dhama K, Prasad M, Ghosh S, Bányai K, Kobayashi N, Singh RK. Multispecies reassortant bovine rotavirus strain carries a novel simian G3-like VP7 genotype.. Infect Genet Evol 2016 Jul;41:63-72.
- Sircar S, Malik YS, Kumar P, Ansari MI, Bhat S, Shanmuganathan S, Kattoor JJ, Vinodhkumar OR, Rishi N, Touil N, Ghosh S, Bányai K, Dhama K. Genomic Analysis of an Indian G8P[1] Caprine Rotavirus-A Strain Revealing Artiodactyl and DS-1-Like Human Multispecies Reassortment.. Front Vet Sci 2020;7:606661.
- Gulati BR, Yadav RR, Singh BK. Epidemiological studies on equine rotavirus infection in foals of organized farms in India. Indian J Anim Sci 79:3–5.
- Khamrin P, Maneekarn N, Peerakome S, Yagyu F, Okitsu S, Ushijima H. Molecular characterization of a rare G3P[3] human rotavirus reassortant strain reveals evidence for multiple human-animal interspecies transmissions.. J Med Virol 2006 Jul;78(7):986-94.
- Khamrin P, Maneekarn N, Peerakome S, Malasao R, Thongprachum A, Chan-It W, Mizuguchi M, Okitsu S, Ushijima H. Molecular characterization of VP4, VP6, VP7, NSP4, and NSP5/6 genes identifies an unusual G3P[10] human rotavirus strain.. J Med Virol 2009 Jan;81(1):176-82.
- Rojas M, Dias HG, Gonçalves JLS, Manchego A, Rosadio R, Pezo D, Santos N. Genetic diversity and zoonotic potential of rotavirus A strains in the southern Andean highlands, Peru.. Transbound Emerg Dis 2019 Jul;66(4):1718-1726.
- Komoto S, Tacharoenmuang R, Guntapong R, Upachai S, Singchai P, Ide T, Fukuda S, Hatazawa R, Sutthiwarakom K, Kongjorn S, Onvimala N, Luechakham T, Sriwanthana B, Murata T, Uppapong B, Taniguchi K. Genomic characterization of a novel G3P[10] rotavirus strain from a diarrheic child in Thailand: Evidence for bat-to-human zoonotic transmission.. Infect Genet Evol 2021 Jan;87:104667.
- Katz EM, Esona MD, Betrapally NS, De La Cruz De Leon LA, Neira YR, Rey GJ, Bowen MD. Whole-gene analysis of inter-genogroup reassortant rotaviruses from the Dominican Republic: Emergence of equine-like G3 strains and evidence of their reassortment with locally-circulating strains.. Virology 2019 Aug;534:114-131.
- Donato CM, Manuelpillai NM, Cowley D, Roczo-Farkas S, Buttery JP, Crawford NW, Kirkwood CD. Genetic characterization of a novel G3P[14] rotavirus strain causing gastroenteritis in 12 year old Australian child.. Infect Genet Evol 2014 Jul;25:97-109.
- Akane Y, Tsugawa T, Fujii Y, Honjo S, Kondo K, Nakata S, Fujibayashi S, Ohara T, Mori T, Higashidate Y, Nagai K, Kikuchi M, Sato T, Kato S, Tahara Y, Kubo N, Katayama K, Kimura H, Tsutsumi H, Kawasaki Y. Molecular and clinical characterization of the equine-like G3 rotavirus that caused the first outbreak in Japan, 2016.. J Gen Virol 2021 Mar;102(3).
- Talmale SS, Pradhan MS. A checklist of valid Indian bat species (Chiroptera: Mammalia). .
- Srinivasulu C, Srinivasulu B. South Asian mammals: their diversity, distribution, and status. .
- Tacharoenmuang R, Komoto S, Guntapong R, Upachai S, Singchai P, Ide T, Fukuda S, Ruchusatsawast K, Sriwantana B, Tatsumi M, Motomura K, Takeda N, Murata T, Sangkitporn S, Taniguchi K, Yoshikawa T. High prevalence of equine-like G3P[8] rotavirus in children and adults with acute gastroenteritis in Thailand.. J Med Virol 2020 Feb;92(2):174-186.
- Luchs A, da Costa AC, Cilli A, Komninakis SCV, Carmona RCC, Boen L, Morillo SG, Sabino EC, Timenetsky MDCST. Spread of the emerging equine-like G3P[8] DS-1-like genetic backbone rotavirus strain in Brazil and identification of potential genetic variants.. J Gen Virol 2019 Jan;100(1):7-25.
- Utsumi T, Wahyuni RM, Doan YH, Dinana Z, Soegijanto S, Fujii Y, Juniastuti, Yamani LN, Matsui C, Deng L, Abe T, Soetjipto, Lusida MI, Ishii K, Shimizu H, Katayama K, Shoji I. Equine-like G3 rotavirus strains as predominant strains among children in Indonesia in 2015-2016.. Infect Genet Evol 2018 Jul;61:224-228.
- Amit LN, Mori D, John JL, Chin AZ, Mosiun AK, Jeffree MS, Ahmed K. Emergence of equine-like G3 strains as the dominant rotavirus among children under five with diarrhea in Sabah, Malaysia during 2018-2019.. PLoS One 2021;16(7):e0254784.
- Bonura F, Mangiaracina L, Filizzolo C, Bonura C, Martella V, Ciarlet M, Giammanco GM, De Grazia S. Impact of Vaccination on Rotavirus Genotype Diversity: A Nearly Two-Decade-Long Epidemiological Study before and after Rotavirus Vaccine Introduction in Sicily, Italy.. Pathogens 2022 Mar 31;11(4).
- Cowley D, Donato CM, Roczo-Farkas S, Kirkwood CD. Emergence of a novel equine-like G3P[8] inter-genogroup reassortant rotavirus strain associated with gastroenteritis in Australian children.. J Gen Virol 2016 Feb;97(2):403-410.
- Gulati BR, Singh BK, Kumar D. Development of a monoclonal antibody-based sandwich ELISA for detection of equine rotavirus from diarrhoeic foals. Indian J Microbiol 46:349–354.
- Gulati BR, Malik P, Kumar R. Isolation and electropherotyping of equine rotaviruses from diarrhoeic foals in India. Indian J Anim Sci 75:745–748.
- Iturriza Gómara M, Wong C, Blome S, Desselberger U, Gray J. Rotavirus subgroup characterisation by restriction endonuclease digestion of a cDNA fragment of the VP6 gene.. J Virol Methods 2002 Aug;105(1):99-103.
- Bolger AM, Lohse M, Usadel B. Trimmomatic: a flexible trimmer for Illumina sequence data.. Bioinformatics 2014 Aug 1;30(15):2114-20.
- Langmead B, Salzberg SL. Fast gapped-read alignment with Bowtie 2.. Nat Methods 2012 Mar 4;9(4):357-9.
- Prjibelski A, Antipov D, Meleshko D, Lapidus A, Korobeynikov A. Using SPAdes De Novo Assembler.. Curr Protoc Bioinformatics 2020 Jun;70(1):e102.
- Zhang KY, Gao YZ, Du MZ, Liu S, Dong C, Guo FB. Vgas: A Viral Genome Annotation System.. Front Microbiol 2019;10:184.
- Pickett BE, Sadat EL, Zhang Y, Noronha JM, Squires RB, Hunt V, Liu M, Kumar S, Zaremba S, Gu Z, Zhou L, Larson CN, Dietrich J, Klem EB, Scheuermann RH. ViPR: an open bioinformatics database and analysis resource for virology research.. Nucleic Acids Res 2012 Jan;40(Database issue):D593-8.
- Edgar RC. MUSCLE: a multiple sequence alignment method with reduced time and space complexity.. BMC Bioinformatics 2004 Aug 19;5:113.
- Tamura K, Stecher G, Kumar S. MEGA11: Molecular Evolutionary Genetics Analysis Version 11.. Mol Biol Evol 2021 Jun 25;38(7):3022-3027.
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