brazilian journal of microbiology49(2018)18–19
h ttp : / / w w w . b j m i c r o b i o l . c o m . b r /
Genome Announcement
Genome sequencing of two Bacillus anthracis strains: a virulent strain and a vaccinal strain
Franciele Maboni Siqueira
a,∗, Samuel Paulo Cibulski
a, Fabiana Quoos Mayer
b, David Driemeier
a, Saulo Petinatti Pavarini
a, Agueda Palmira Castagna de Vargas
caUniversidadeFederaldoRioGrandedoSul(UFRGS),FaculdadedeVeterinária,DepartamentodePatologiaClínicaVeterinária,Porto Alegre,RS,Brazil
bFundac¸ãoEstadualdePesquisaAgropecuária,InstitutodePesquisasVeterináriasDesidérioFinamor(IPVDF),LaboratóriodeBiologia Molecular,EldoradodoSul,RS,Brazil
cUniversidadeFederaldeSantaMaria(UFSM),CentrodeCiênciasRurais,DepartamentodeMedicinaVeterináriaPreventiva,Santa Maria,RS,Brazil
a r t i c l e i n f o
Articlehistory:
Received13March2017 Accepted27April2017 Availableonline31July2017 AssociateEditor:JohnMcCulloch
Keywords:
Anthrax
Vaccinepathogenicity Bovineabortion
Nextgenerationsequencing
a bs t r a c t
BacillusanthracisstrainSPV84215wasisolatedfrombovinefetus,whileB.anthracisstrain Brazilianvaccinalwasrecoveredfromacommercialvaccine.Wereportherethegenome sequencesofbothstrains.TheSPV84215genomeiscomposedofasinglecircularchromo- somewithalengthof5,228,664basepairs,andcomprises5911codingsequences.Inturn, theBrazilianvaccinalgenomeremainsin201contigswith5733codingsequences.Both genomeshaveanoverallC+Gcontentof35.4%,and11genesencodingtheribosomalRNAs (rRNAs)5S,16Sand23S.OnlytheplasmidpX01sequence,whichcarriesgenesfortoxins synthesis,wasdetectedandcompletelyassembledforbothstrains.Theseplasmidshave alengthof181,684basepairsandaC+Gcontentof32.5%.Thesegenomicdatagenerate insightsaboutvaccinalB.anthracisvirulence.
©2017SociedadeBrasileiradeMicrobiologia.PublishedbyElsevierEditoraLtda.Thisis anopenaccessarticleundertheCCBY-NC-NDlicense(http://creativecommons.org/
licenses/by-nc-nd/4.0/).
Genome announcement
Bacillus anthracis is a Gram-positive bacterium that causes anthrax.1 The anthrax veterinary vaccine is anattenuated non-encapsulatedandtoxigenicB.anthracisstrain.2Theatten- uationisduetothelossofthepXO2plasmid,whichcarries
∗ Correspondingauthor.
E-mail:[email protected](F.M.Siqueira).
thegenesrequiredforthebiosynthesisanddegradationofan anti-phagocyticcapsule.3Ontheotherhand,thepXO1plas- midcarriesthegenesforthelethalfactor,theedemafactor, andaprotectiveantigen,4andispresentinbothpathogenic andvaccinalB.anthracisstrains.
Herewedescribetwogenomesequences:(i)thegenome of B. anthracis strain SPV84215, which was isolated from AberdeenAngusbovinefetuses,and(ii)thegenomeofavac- cinestrain, which wenamedB.anthracis Brazilianvaccinal strain.
https://doi.org/10.1016/j.bjm.2017.04.007
1517-8382/©2017SociedadeBrasileiradeMicrobiologia.PublishedbyElsevierEditoraLtda.ThisisanopenaccessarticleundertheCC BY-NC-NDlicense(http://creativecommons.org/licenses/by-nc-nd/4.0/).
brazilian journal of microbiology49(2018)18–19
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TotalDNAfromthebacteriastrainswereextractedbylysis withcetyltrimethylammoniumbromide(CTAB)andisolation bytheadditionofphenol:chloroform.DNAlibrarieswerepre- pared with Nextera DNA Library Kit and high-throughput sequencingwasperformedonIlluminaMiSeqsystem (Illu- mina,Inc.)togenerate150-bppairedendreads(v2chemistry) accordingtothemanufacturer’sinstructions.
Reads were imported into the Geneious software (ver- sion 9.1.5) and trimmed. Assembling of the genomes was thenaccomplishedbydenovoassemblyusingSPAdes3.9.05 followedby template-assistedassemblyto the referenceB.
anthracisSternestrain(NC005945)andB.anthracisAmesstrain (NC003997). NCBI ProkaryoticGenome AnnotationPipeline performedtheannotationsofthegenomes.
Thewholegenomesequencinggenerated11xand23xcov- erage,forBrazilianvaccinalgenomeandSPV84215genome, respectively.BothgenomeshaveanoverallC+Gcontentof 35.4%.TheSPV84215genomeiscomposedofasinglecircu- larchromosomewithalengthof5,228,664bps,andcomprises 5911CodingSequences(CDSs).Inturn,theBrazilianvaccinal genomeremainsin201contigswith5733CDSs.Eachgenome contains11genesencodingtheribosomalRNAs(rRNAs)5S, 16Sand23S,and95genesencodingthetransferRNAs(tRNAs).
Inbothstrains,onlytheplasmidpX01sequencewasdetected andcompletelyassembled.Theseplasmidshavealengthof 181,684bps.
ThepXO2plasmidwasnotdetectedintheSPV84215orin theBrazilianvaccinalstrains.Therefore,wehypothesizedthat thevaccinenon-encapsulatedattenuatedbacteriamayhave apathogenicthreshold.
ThecompletesequencesofthepX01plasmidfromeachB.
anthracisgenomeswereassembledandthealignmentshowed ahighidentitylevel.Thereareentireidentitiesinthewhole pX01plasmidsequences.Accordingtotheassembly,thepX02 plasmid isabsent in the pathogenic B. anthracisSPV84215 strain,andthepX01plasmidispresentwithtotalidentityto thepX01 plasmidfrom the Brazilianvaccinalstrain. These genomic datagenerate insights about thepathogenicity of
B.anthracis.Additionalcomparativegenomicstudiesinthese strainswillhelptounderstandthepotentialinfectionprofile ofB.anthracisvaccine,andprovideinformationtounderstand thesourcedynamicsoftheB.anthracisSPV84215strain.
The genomes and pX01 plasmid sequences have been deposited inDDBJ/ENA/GenBankunderthe accessionnum- bers CP019588.1 and CP019589.1 for B. anthracis SPV84215 genomeandplasmid,respectively;andMVOA00000000forB.
anthracisBrazilianvaccinalShotgungenome.
Conflicts of interest
Theauthorsdeclarenoconflictsofinterest.
Acknowledgments
WewouldliketothanktoFundac¸ãodeApoioàTecnologiae ciênciadaUniversidadeFederaldeSantaMaria(FATEC/UFSM) –proccess5.03.0017–forthefinancialsupport.
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