Posted on Authorea 22 May 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159018071.14311072 — This a preprint and has not been peer reviewed. Data may be preliminary. eeOrtega Rene a in infection guigna) gui˜na 1 (Leopardus free-ranging Protoparvovirus rescued Carnivore of origin dog Domestic osaz Napolitano Constanza Chile. 2 Concepci´on, Chill´an, Chile. 1 Affiliations: authors *Co-corresponding Cofr´e Napolitano Authors: Head: Running guigna) pardus Title: this of programs a protection gui˜na is The for that conservation. account its into highlight taken to findings thread be studies gui˜na current important should comparative and The an species. dogs few represent endangerous dogs. the domestic dogs the between domestic is domestic of transmission were This that cross-species one and infection L582F. CPV infection and CHL-guigna of protoparvovirus gui˜na worldwide and source to in E661K species the infection susceptible CHL-71 that presented CPV-2c V596A, determine viruses of CHL-17 that These undoubtedly characterization showed protein, strains. that genetic genome NS1 dogs and the coding domestic detection in complete Chilean we changes first the with three Here, ancestor of showed common analysis dogs carnivores. and recent domestic phylogenetic identity domestic a and and and guigna) share CPV-2c gui˜na (Leopardus strain wild rescued as gui˜na CPV-2c both a classified the from affecting was strains Gui˜na pathogens sequence parvovirus Chile. important canine from most of characterization the genetic of the one reported is 1 protoparvovirus Carnivore Abstract 2020 22, May 6 5 4 3 2 1 Gonz´alez-Acu˜na nvriyo Concepci´on of University Concepci´on de Universidad Lagos Los de Universidad Rep´ublica Uruguay la de Universidad Chile de Universidad Concepcion de Universidad eatmnod ini nml autdd inisVtrnra,Uiesddd Concepci´on, Chill´an, de Universidad Veterinarias, Ciencias de Facultad Animal, Ciencia de Departamento eatmnod aoo´ayMdcn rvnia autdd inisVtrnra,Uiesddde Universidad Veterinarias, Ciencias de Facultad Preventiva, Medicina Patolog´ıa y de Departamento 3 V´ıctor Neira , oetcdgoii fCrioePooavvrs1ifcini ece rerniggui˜na ( free-ranging rescued a in infection 1 Protoparvovirus Carnivore of origin dog Domestic 5,6 e´ Ortega Ren´e hrAsh Zegpi Nhur-Aischa 1 unMn Vasquez Mena Juan , P-cifcini gui˜na in infection CPV-2c 5 egoCofr´e Sergio , 4 n Crist´obal Castillo-Aliaga and 1 unMena Juan , 4 hrAsh Zegpi Nhur-Aischa , 1 let Sandoval Alberto , 1 itrNiaRamirez Neira victor , 4* 2 o´aGrecco Sof´ıa , Sof´ıa Grecco , 1 1* let Sandoval Alberto , 1 . 1 ailSandoval Daniel , 7 u´nP´erez Rub´en , 3 ue P´erez Ruben , 2 n rsoa Castillo-Aliaga Cristobal and , 1 ailGonz´alez-Acu˜na Daniel , 1 7 ailSandoval Daniel , aiaPanzera Yanina , 3 aiaPanzera Yanina , 7 Constanza , 1 Daniel , 3 , 2 Sergio , 6 > Leo- 99% Posted on Authorea 22 May 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159018071.14311072 — This a preprint and has not been peer reviewed. Data may be preliminary. eogn otodffrn iegs(P´erez (Castillo lineages origin different that from European two lineage of genome to strains a belonging 2c complete and of Asia of predominance from the analysis introduction with an The (Grecco Europe, South-America described. in from high diverged events were likely migration The variants two years. antigenic evidenced countries few and (Truyenseveral species genetic in non-dog worldwide South-America, for In spread thread antigenic/genetic a and the including pose felids (Shackelton variants, to 2c genetic infect started y new and to 2b of 2a, ability emergence species as the the known carnivore promoted variants wild regained CPV related of that a capacity or lineage mutation felids new infected a likely that by ancestor recent common a (Truyen non-carnivore share a CPV in and infection FPV CPV-2 of evidence first the order providing (Wang the pangolin, species Taiwanese in rescued animals a in domestic described and wild mink Chen both (CPV), 2019; parvovirus affect (Cotmore encodes canine and parvovirus ORF relevance (FPV), second raccoon veterinary parvovirus the and feline while virus, including NS2), enteritis species, (Cotmore and different (ORFs) (Reed (NS1 considered VP2 frames proteins and reading previously non-structural VP1 open two proteins major structural encodes single-stranded two the linear, ORF containing a first kb having The viruses 5.2 2014). icosahedral approximately small of non-enveloped, genome are DNA these parvoviruses, other Like . 1 protoparvovirus Carnivore Introduction Text Main species. be gui˜na endangerous important should and this an dogs of Keywords: represent domestic programs dogs between protection transmission domestic that for cross-species highlight account that CPV findings into and The current taken and infection conservation. The detection protoparvovirus its dogs. first to to domestic that thread the were studies species comparative infection is susceptible few of This a the source gui˜na L582F. of is the one CHL-guigna that and gui˜na and determine worldwide in undoubtedly E661K infection CPV-2c CHL-71 presented of V596A, characterization viruses genetic CHL-17 ancestor These protein, common recent strains. NS1 a dogs share the gui˜na strain domestic gui˜na the CPV-2c that rescued Chilean showed a genome with from coding complete strains the parvovirus of canine analysis of characterization genetic the ( reported we Here, carnivores. 1 protoparvovirus Carnivore Summary Rep´ublica, Uruguay. la de Universidad 7 6. 5 Chile. Santiago, Chile, de 4 Chile. 3 epru guigna Leopardus eatmnod inisC´ncs autdd inisVtrnra,Uiesddd Concepci´on, Chill´an, de Universidad Veterinarias, Ciencias Cl´ınicas, de Facultad Ciencias de Departamento ec´ nGneiaEouia eatmnod ilgı nml nttt eBoo´a autdd Ciencias, Biolog´ıa, de Facultad de Instituto Biolog´ıaSecci´on Animal, de Gen´etica Departamento Evolutiva, eatmnod eiiaPeetv nml autdd inisVtrnra eura,Universidad Pecuarias, y Veterinarias Ciencias de Facultad Animal, Preventiva Medicina de Departamento eatmnod inisBo´gcsyBoiesdd nvria eLsLgs sro Chile. Osorno, Lagos, Los de Universidad Biodiversidad, Biol´ogicas y Ciencias de Departamento nttt eEoo´ayBoiesdd(E) atao Chile. Santiago, (IEB), Biodiversidad Ecolog´ıa y de Instituto tal et tal et aieparvovirus, Canine ,19) hnCV2eegd tol netdcnd.Ti rtCV2tp a onreplaced soon was type CPV-2 first This canids. infected only it emerged, CPV-2 When 1992). ., tal et ,21;Cotmore 2019; ., n oetcdg rmCie u˜asqec a lsie sCV2 n phylogenetic and CPV-2c as classified Gui˜na was Chile. sequence from dogs domestic and ) ,2019). ., CP-)i ebro h genus the of member a is (CPPV-1) soeo h otipratptoesaetn ohwl n domestic and wild both affecting pathogens important most the of one is epru guigna Leopardus tal et ,21;Cotmore 2014; ., tal et tal et tal et tal., et ,21) hl,UuuyadAgniahv iia scenario similar a have Argentina and Uruguay Chile, 2018). ., ,20) hs ainsrcvrdteaiiyt netfelines infect to ability the recovered variants These 2005). ., ,21;Gallo-Calder´on 2014; ., rtproiu ,pyoey eei characterization genetic phylogeny, 1, protoparvovirus , tal., et 98.CP- opie eea lsl-eae lineages closely-related several comprises CPPV-1 1988). 2 tal et 04.Teevrssaesgicn ahgn of pathogens significant are viruses These 2014). tal et ,21) aa P- neto a recently was infection CPV-2 fatal A 2019). ., > ,19;Ikeda 1996; ., 9 dniyadsoe he hne in changes three showed and identity 99% tal et Protoparvovirus ,22)adteeitneo astrains 2a of existence the and 2020) ., tal et ,2015). ., tal et Carnivore ftefamily the of ,2000). ., (Behdenna Parvoviridae tal et tal., et ., Posted on Authorea 22 May 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159018071.14311072 — This a preprint and has not been peer reviewed. Data may be preliminary. ttsia upr ftendswseautduig10 otta replicates. bootstrap 1000 discussion using and with evaluated Results method jModelTest was (Miller Gateway of likelihood nodes in Science maximum the model (AIC) CIPRES the of evolutionary in Criterion using support available statistical inferred best-fit Information 2014) was Akaike The (Stamatakis, tree v8.2.12 MUSCLE the 2014). Phylogenetic using RaxML-HPC2 to 2008). (Larsson, aligned (Posada, were according v1.26 software sequences selected sequences v2.1.6 AliView The was genome dog. in than substitution coding incorporated other subtype, nucleotide complete its species 2004) to CPV host according (Edgar, sequences and and the collection v3.8.3 select study of to was year this criterion and filter in country The generated database. GenBank sequences following the from amplified of retrieved gui˜na were consisted and dogs dataset from The pb) (4269 Buonavoglia genome genome following coding coding complete amplified complete Castillo P´erez for the was by selected Therefore, characterized bp) were gene. previously (583 dogs VP2 were VP2 domestic samples Chilean of These from fragment sequencing. samples viral Chile seven small Stool Furthermore, of DNA University a (2001). -20 QIAamp Sciences, at diagnosis, the of stored CPV Faculty were and For Laboratory, DNAs Kit Biology genomic Tissue Molecular extracted and the The Blood Chile)). at DNeasy (Santiago, (Processed using Qiagen samples Kit; tissue Concepci´onMini from (Chill´an, of University Chile)). extracted DNA Sciences, spleen Veterinary was Scientific of A and DNA Faculty ThermoFisher Laboratory, intestine FIV. using Virology small sample the and at FeLV stomach, blood cessed to from Stool, negative extracted out. was carried and DNA was CPV necropsy (Mora for and PCR positive to collected. died, Nested resulting collected were animal by was tissues performed, the FIV sample were later, FelV, PCR weeks blood against by few A test the CPV exam. Diagnostic by against clinical cause. Curic´o (Chile) and the infectious intermittent from during an possible area presented anorexia a individual urban identify and The Concepci´on Rehabilitation (Chill´an, depression an Chile). de for Center diarrhea, Universidad in the the hemorrhagic to recovered submitted of was gui˜na was Fauna animal Wild The sick Chile. of six-month-old of (SAG) Service a Livestock phylogenetic and 2015, Agricultural the of establish July area. to geographic In gui˜na and near in the CPV inhabiting Methods the dogs and domestic characterize Materials from to obtained was strains study CPV (Mora this viruses with phylogenetic of relationship animals cases, domestic objective both and main in wild The and between gui˜na populations association feline in high and reduction (FIV) reported a immunodeficiency the suggested been viral are have analyses feline causes years, infections main recent (FeLV) In the leukaemia 2015). and (UICN, viral vulnerable, habitat is its status of fragmentation conservation and its (IUCN), Nature of Conservation sn P euneaayi n vdne h xsec fapeoiat2 ain fErpa origin European of (Napolitano variant Argentina Two 2c southwestern predominant animals. a canids wild gui˜na ( of wild The and existence and dogs the dogs domestic evidenced domestic and both in (Castillo analysis in CPV-2 sequence CPV against VP2 antibodies of using of circulation presence the the (Acosta-Jammet on evidenced information studies little serological a has Chile ( coati dead a from characterized 2019). genetically and isolated was Curi nwl pce rmSuhAeia hr satgnceiec ftecruaino PV1i idanimals, wild ( in foxes CPPV-1 of crab-eating circulation the and of (Martino evidence grey antigenic culpeo, is there including South-America, from species wild In tal et tal et tal et tal et ,21)adGorysct ( cats Geoffrey’s and 2012) ., ,(2014). ., epru guigna Leopardus ,2020). ., ,20;Acosta-Jammet 2004; ., tal et ,21;Acosta-Jammet 2014; ., sasalfldihbtn eta n otenCieadsm ra in areas some and Chile southern and central inhabiting felid small a is ) tal et epru geoffroyi Leopardus tal et ,21) codn oterdls fteItrainlUinfor Union International the of list red the to According 2014). ., ,21) ae ovs( wolves maned 2014), ., tal et yaoe culpaeus Lycalopex ,21) oercn td hrceie h strains the characterized study recent more A 2015). ., 3 (Uhart ) au nasua) Nasua ; tal et ° C. tal et yaoe rsu;Croynthous Cerdocyon griseus; Lycalopex hyoynbrachyurus Chrysocyon ,21) eety P-cstrain CPV-2c a Recently, 2012). ., ,(00 sn eunigo the of sequencing using (2020) ., ® nAgnia(Bucafusco Argentina in xrcinfo lo (pro- Blood from Extraction tal et tal et ,2015). ., d Almeida (de ) ,21) The 2010). ., tal., et tal et tal et 2015) ., ., ) Posted on Authorea 22 May 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159018071.14311072 — This a preprint and has not been peer reviewed. Data may be preliminary. hr r o aysuista hwsqecscmaio n nlssbtendmsi osadwild and dogs domestic between analysis (Steinel and GenBank comparison (Wang sequences in CPV show to that (Chen only than studies the species rather many been not FPV has are to this There similarity and greater China, showing in felid, tigers wild captive (Bucafusco 2019). strain in a coati described from the been described with has variant described (TPV) was parvovirus situation similar tiger Recently, and strains, dog domestic with Allison h uhr elr ocnito interest of conflict no declare authors Programa The the RO, statement to interest 217.152.024-1.0 number of UdeC Conflict Inicio, VN VRID to project 11170877 Iniciaci´on by No. de funded Fondecyt partly was study This Funding especially to Pathology, especially Animal cases. sampling, of clinical processing Department providing initial of the Laboratory in Diagnostic support Molecular their to the all thank for We FISENLAB of Rojas. these staff Pedro by the infection to grateful of are risk We permanent at be would from animals agents Acknowledgments gui˜nas these infectious (Mora in that of described suggest transmission previously might the been finding consider pathogens. has our to FIV So, important and FeLV 2014). be by ., will infection Likewise, it animals. species, domestic this of conservation the in reported been have felids wild wildlife jubatus in for Acinonyx affinity infections greater CPV a according have Several may clade lineage Europe-I 2c the (Bucafusco the apparently in South-America Furthermore, in clustered dogs. Strains Grecco both domestic 1). by of but (Figure acquisition classification sub-clades, Paraguay recent different and the in Argentina, Uruguay, to diverged Chile, coati from gui˜na and (P´erez 2009-2017 from variant between CPV-2a collected (Castillo unrelated strains variant phylogenetically dog 2c a by of replaced predominance being Gallo-Calder´on a is have variant different Argentina this was Uruguay and associated residue Chile the South-America, but In strains FPV position three NS1 of in 582 protein tropism the NS1 tissue in (Garigliany the neurological Changes (L582S) in 3). with (Table changes associated L582F to CHL-guigna been lead and has high polymorphism E661K nucleotide showed CHL-71 (Buonavoglia V596A, single sequences amino- CHL-17 Three strains showed dog samples, gui˜na 2). sequence 2c (Table the Chilean 426E of gui˜na other and and region The with 300G VP2 1. the associated Table of changes in analysis acid sequence shown domestic The are from (99.8-99.9%). gui˜na seven sequences identity the and from genetic these one regions, of coding Details CPV found the dogs. those of sequences to complete similar eight obtained were We case this haemor- in (Allison The Rehabilitation efficiently. collected species Bucafusco for host other tissues 2019; the Center in infected in the infections the viremia protoparvovirus to of established in arrival characteristic and and infected the virus diarrhoea at the rhagic PCR that by suggesting positive and diagnosis CPV Fauna. gui˜na was the Wild of of sample blood The laoRi,fralterspoti rvdn hi aiiis etakD.FacsoEpn oe for Lopez Espina Francisco Dr. thank We facilities. their providing in support their all for Ruiz, Alvaro ´ tal et ,21)and 2014) ., tal., et tal et Post-mortem tal et , ateatigris Panthera tal et ,21) hlgntcaayi hwdta h u˜aproiu lsee ihCPV-2c with gui˜na clustered parvovirus the that showed analysis Phylogenetic 2015). ., 09 Oosthuizen 2019; tal et ,21;Wang 2019; ., 2016). , etiuu serval Leptailurus ,20;Nakamura 2000; ., tal et to,soahadsle ape eeas oiie ofimn h CPV the confirming positive, also were samples spleen and stomach stool, (Steinel ,2019). ., tal et tal et tal et ,21;Viscardi 2019; ., tal et 21) u eut,sget httegin iu sarecent a is gui˜na virus the that suggests results, Our (2018). . (Oosthuizen ,21) otrprsol opr ihsqecsavailable sequences with compare only reports Most 2019). ., tal., et ,2000), ., 4 01 Allison 2001; tal et uaconcolor Puma tal et tal et ,21;Allison 2013; ., tal et ei bengalensis Felis ,2019). ., ,21) l fte hwdhg identity high showed them of All 2019). ., ,20;Decaro 2001; ., tal., et , yxrufus Lynx 03 Allison 2013; tal et ,21;Oosthuizen 2014; ., (Nakamura tal et (Allison tal et tal et ,21) including 2012), ., ,22) u in but 2020), ., tal et tal et ,21) For 2014). ., tal et tal et ,2001), ., ,2019). ., ,2013; ., ,2014; ., tal et tal et tal et ., ., Posted on Authorea 22 May 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159018071.14311072 — This a preprint and has not been peer reviewed. Data may be preliminary. n isesmlswr ae ntencos fe h death. the after procedure necropsy clinical the a References owners in during the taken taken was were from gui˜na sample The samples obtained de individually. not tissue Universidad identified Veterinarias, was and not Ciencias participation were animals for de Facultad the consent received. because la informed been de written page, has was Bioetica guidelines approval Comit´e samples de author Concepci´on. by committee Dog journal’s approved review the and ethical on reviewed appropriate noted Was as the journal, and the to of numbers adhered policies Accession been ethical have the at that GenBank confirm authors in The available openly MT458230. are and study Statement MT458229 Ethics this MT458228, of MT458227, MT458226, findings MT458225, the MT458224, support MT458223, that data The Statement Availability Data Dcr,N,&Boaola .(02.Cnn avvrsarve feieilgcland epidemiological of review parvovirus—a Canine (2012). C. 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Torre, Microbiology, La Veterinary https://doi.org/10.1093/nar/gkh340 1792-1797. 2c. 32(5), research, acids type Nucleic on emphasis with ps://doi.org/10.1016/j.vetmic.2011.09.007. aspects, diagnostic 6 Posted on Authorea 22 May 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159018071.14311072 — This a preprint and has not been peer reviewed. Data may be preliminary. otdfile Hosted leopardus-guigna origin-of-carnivore-protoparvovirus-1-infection-in-a-rescued-free-ranging-gui%C3%B1a- Table_2.docx file Hosted leopardus-guigna origin-of-carnivore-protoparvovirus-1-infection-in-a-rescued-free-ranging-gui%C3%B1a- colors. different in domestic represented Table_1.docx Chilean are clades. strains main CPV the species tree file in other phylogenetic Hosted indicated and The are dog, substitution nucleotide values GenBank. domestic GTR+I+G4 reference Bootstrap in on available guina, replicates. based dog, sequences RaxML bootstrap in CPV 1000 method with all likelihood model and maximum the study using this inferred was in obtained sequences the 1 Figure 3. Table 2. Table included. were type virus city, collection, 1. Table Legends Table and Figure Wn,K,D,S,Wn,Y,Wn,S,Lo . hn,Y,...&H,G 21) slto and Isolation (2019). G. Hu, & . . . Y., Zhang, X., Luo, canine Fatal S., (2019). Wang, T. W. Y., Li Wang, & , S., . . . Du, H., K., C. Wu Wang, Y., T. 38. Chen H., L. Wu S., G. M. Fusco, Lee C., & Y. Tu L., L., M. S. Wang Colaianni, 37. N., Decaro, L., Cozzolino, T., M. Clausi, M., selected to Santoro, Exposure M., (2012). A. Viscardi, J. Pereira 36. V., del H. Ferreyra A., C. Marull, V., involved M. parvovirus Rago, feline canine M., of M. and Evolution Uhart, (1996). parvovirus R. C. 35. canine Parrish, & of E., Vieler, ranges F., fe- J. host Evermann, U., feline of Truyen, and characterization 34. Canine Genetic (1992). R. (2000). C. U. Parrish, Truyen, & & U., Truyen, M., 33. large Vuuren, of van post-analysis and L., analysis Munson, phylogenetic for A., tool Steinel, a as- 8: 32. evolution version viral RAxML of (2014). rate High A. (2005). Stamatakis, C. 31. E. Holmes, & U., Truyen, R., C. Parrish, A., L. Shackelton, 30. dnicto ftgrproiu ncpiesbra iesadpyoeei nlsso P gene. VP2 of analysis phylogenetic and https://doi.org/10.1016/j.meegid.2019.103957 tigers 103957. siberian 75, captive Evolution, and in Genetics parvovirus Infection, tiger https://doi.org/10.1111/tbed.13469 of 30. identification Dec 2019 1864-1872. pen- pentadactyla Disease. 66(5), (Manis Emerging pangolin Taiwanese and diseases, Transboundary free-ranging rescued tadactyla). emerging a in and infection (CPV-2) Transboundary parvovirus-2 Eurasian Italy. free-ranging in southern parvoviruses in carnivore https://doi.org/10.1111/tbed.13212 lutra) of characterization (Lutra and otters detection Argentina. Molecular central (2019). https://doi.org/10.7589/2011-05-137. from 909. carnivores 899- domestic and 48(4), cats Disease, Geoffroy’s Wildlife in of pathogens Journal selected to 186-189. in 215(2), pathogens Virology, range. virology, host of feline Journal of vivo. gain in and and loss vitro in 345-350. virus each 81(2), of Virology, tropisms 5399-5408. cell 66(9), General host distinct of virus: panleukopenia Journal carnivores. various from https://doi.org/10.1099/0022-1317-81-2-345 sequences parvovirus line 10.1093/bioinformatics/btu033 https://doi.org/1312-1313. 30(9), Bioinformatics, phylogenies. Sciences, of Academy National the of Proceedings https://doi.org/10.1073/pnas.0406765102 parvovirus. 379-384. carnivore 102(2), of emergence the with sociated mn cdvrain nN1i ape nlsdi hsstudy. this in analysed samples in study. NS1 this in in variations analysed acid samples Amino in VP2 in variations acid Amino hlgntcaayi fteCVcmlt oigrgo.Tedtbs a osrce using constructed was database The region. coding complete CPV the of analysis Phylogenetic . apeietfiainue nti td.Ietfiaino ceso ubr tannm,date name, strain number, accession of Identification study. this in used identification Sample vial at available at available https://authorea.com/users/325377/articles/453323-domestic-dog- https://authorea.com/users/325377/articles/453323-domestic-dog- 7 Posted on Authorea 22 May 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159018071.14311072 — This a preprint and has not been peer reviewed. Data may be preliminary. leopardus-guigna origin-of-carnivore-protoparvovirus-1-infection-in-a-rescued-free-ranging-gui%C3%B1a- Table_3.docx vial at available https://authorea.com/users/325377/articles/453323-domestic-dog- 8