LETTERS

The molecular masses of the PrPres Prionics AG, and the veterinary services 5. Offi ce Internationale des Epizooties. OIE moieties from the 2 cows were also of the cantons of St. Gallen and Berne rules for the offi cial confi rmation of BSE in bovines (based on an initial reactive clearly distinct from those from for their support. We also thank Jan P.M. result in an approved rapid test) by using controls with L- and H-BSE (Figure). Langeveld for kindly providing antibodies a second rapid test. 2009 [cited 2011 Oct For samples from animals with 9A2 and 94B4. 19]. http://vla.defra.gov.uk/science/docs/ H-BSE, enzymatic deglycosylation sci_tse_oie_bse.pdf This work was fi nanced with 6. Biacabe AG, Jacobs JG, Bencsik A, demonstrated PrPres subtypes, 1 and resources provided by the Swiss Federal Langeveld JP, Baron TG. H-type bovine 2, the latter being a C-terminal PrPres Veterinary Offi ce. spongiform encephalopathy: complex fragment of ≈12–14 kDa (6). To molecular features and similarities with investigate whether the novel PrPres human prion diseases. Prion. 2007;1:61– 8. doi:10.4161/pri.1.1.3828 type corresponds to PrPres subtype Torsten Seuberlich, 2, we compared samples from cow Michaela Gsponer, Cord Drögemüller, Address for correspondence: Torsten 2 with those from the H-BSE control Seuberlich, NeuroCentre, National and OIE res Miroslaw P. Polak, by Western blot. The PrP type from Reference Laboratories for BSE and Scrapie, res Sandra McCutcheon, the 2 cows reported here and PrP University of Berne, Bremgartenstrasse 109a, subtype 2 from the H-BSE control Dagmar Heim, Anna Oevermann, CH- 3001 Berne, Switzerland; email: torsten. were indeed distinct (Figure). [email protected] We report a novel PrPres signature and Andreas Zurbriggen in 2 cows with BSE diagnoses Author affi liations: University of Bern, Bern, determined according to established Switzerland (T. Seuberlich, M. Gsponer, C. criteria. Combining Western blot Drögemüller, A. Oevermann, A. Zurbriggen); analysis with an epitope mapping National Veterinary Research Institute, strategy, we ascertained that these Pulawy, Poland (M.P. Polak); University animals displayed an N terminally of Edinburgh, Edinburgh, Scotland, UK truncated PrPres different from (S. McCutcheon); and Federal Veterinary Hantavirus in Bat, currently classifi ed BSE prions Offi ce, Liebefeld, Switzerland (D. Heim) Sierra Leone (Figure). The interpretation of these DOI: http://dx.doi.org/10.3201/eid1801.111225 fi ndings remains diffi cult because To the Editor: Hantaviruses neuropathologic and systematic References (family Bunyaviridae) are transmitted clinical data for the 2 cases are not from rodent reservoirs to humans. available. Moreover, the tissue 1. Colby DW, Prusiner SB. Prions. Cold These viruses cause life-threatening Spring Harb Perspect Biol. 2011; samples were autolyzed, and the 3:a006833. doi:10.1101/cshperspect.a00 human diseases: hantavirus question of whether this affected 6833 cardiopulmonary syndrome in the the PrPres molecular signature is of 2. Jacobs JG, Langeveld JP, Biacabe AG, Americas and hemorrhagic fever concern. Nonetheless, our fi ndings Acutis PL, Polak MP, Gavier-Widen D, with renal syndrome in Asia and et al. Molecular discrimination of atypi- raise the possibility that these cattle cal bovine spongiform encephalopathy Europe (1). Since 2006, indigenous were affected by a prion disease not strains from a geographical region span- hantaviruses were reported also from previously encountered and distinct ning a wide area in Europe. J Clin Mi- Africa. Sangassou virus was found in from the known types of BSE. To crobiol. 2007;45:1821–9. doi:10.1128/ an African wood mouse (Hylomyscus JCM.00160-07 confi rm this possibility and to assess a 3. Seuberlich T, Heim D, Zurbriggen A. simus) in Guinea (2). Discovery of potential effect on disease control and Atypical transmissible spongiform en- newer African hantaviruses, Tanganya public health, in vivo transmission cephalopathies in ruminants: a challenge virus and recently Azagny virus, was studies using transgenic mouse for disease surveillance and control. even more surprising because they J Vet Diagn Invest. 2010;22:823–42. models and cattle are ongoing. Until doi:10.1177/104063871002200601 were found in shrews (3,4). results of these studies are available, 4. Schaller O, Fatzer R, Stack M, Clark J, The detection of hantaviruses in molecular diagnostic techniques Cooley W, Biffi ger K, et al. Validation of small mammals other than rodents, should be used so that such cases are a Western immunoblotting procedure for such as shrews and also moles (4), bovine PrP(Sc) detection and its use as a not missed. rapid surveillance method for the diagno- increasingly raises questions regarding sis of bovine spongiform encephalopathy the real hantavirus host range. Acknowledgments (BSE). Acta Neuropathol. 1999;98:437– Bats (order Chiroptera) are already We thank the BSE screening 43. doi:10.1007/s004010051106 known to harbor a broad variety of laboratories at the Center of Laboratory emerging pathogens, including other Medicine (ZLM, St. Gallen, Switzerland), bunyaviruses (5). Their ability to fl y

Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 18, No. 1, January 2012 159 LETTERS and social life history enable effi cient bat-borne hantavirus with shrew- from those of other hantaviruses, we pathogen maintenance, evolution, and mole-associated hantaviruses conclude that the bat was infected and spread. Therefore, we conducted seems reasonable. Notably, shrew- with a newly found hantavirus. a study on hantaviruses in bats from associated Thottapalayam virus We propose the putative name Africa. (India) and Imjin virus (South Korea) Magboi virus (MGBV) for the new A total of 525 tissue samples seem to be closer relatives, and virus because it was detected in from 417 bats representing 28 genera African Tanganya virus (Guinea) an animal captured at the Magboi were tested for the presence of and Azagny virus (Côte d’Ivoire) are River in Sierra Leone. The MGBV hantavirus RNA. Samples originated more distantly related. Additional nucleotide sequence is novel and has from different regions in western and sequence data is needed for more not been known or handled before central Africa and were collected conclusive phylogenetic analyses. in our laboratory. Before this study, during 2009 and early 2011. Total RNA Because the new amino acid hantavirus nucleic acid was found in was extracted from tissue samples sequence is at least 22% divergent lung and kidney tissues of bats from and reverse transcribed. cDNA was screened by PCR specifi c for sequences of the large genomic segment across the genus Hantavirus (2). One sample yielded a product of the expected size and was subjected to cloning and sequencing. The positive sample (MGB/1209) was obtained from 1 of 18 investigated slit-faced bats (family Nycteridae). The animal was trapped at the Magboi River within Gola National Park, Sierra Leone (7°50.194′N, 10°38.626′W), and the identifi cation as Nycteris hispida has been verifi ed with the voucher specimen (RCJF529). Histologic examination of organs of the animal showed no obvious pathologic fi ndings. The obtained 414-nt sequence covers a genomic region, which was found to correspond to nt position 2,918–3,332 in the large segment Figure. Maximum-likelihood phylogenetic tree of MGB/1209 virus based on partial open reading frame of prototypic large segment sequence (414 nt) and showing the phylogenetic placement of the novel Hantaan virus. Bioinformatic analysis sequence from Nycteris spp. bat compared with hantaviruses associated (i) with shrews on the amino acid level showed and moles: Altai virus (ALTV), Artybash virus (ARTV), Asama virus (ASAV), Ash River virus highest degrees of identity to shrew- (ARRV), Azagny virus (AZGV), Camp Ripley virus (RPLV), Cao Bang virus (CBNV), Imjin and mole-associated hantaviruses virus (MJNV), Jemez Springs virus (JMSV), Kenkeme virus (KKMV), Nova virus (NVAV), Oxbow virus (OXBV), Seewis virus (SWSV), Tanganya virus (TGNV), Thottapalayam virus (Thottapalayam virus 73.0%, (TPMV), and (ii) with rodents: Andes virus (ANDV), Choclo virus (CHOV), Dobrava-Belgrade Altai virus 69.7%, Nova and Imjin virus (DOBV), Hantaan virus (HTNV), Laguna Negra virus (LNV), Maporal virus (MAPV), virus 69.3%). On the basis of tree Puumala virus (PUUV), Rio Mamore virus (RIOMV), Sangassou virus (SANGV), Seoul topology of a maximum-likelihood virus (SEOV), Serang virus (SERV), Sin Nombre virus (SNV), Soochong virus (SOOV), phylogenetic tree, the sequence does Tula virus (TULV), Vladivostok virus (VLAV). The list of the accession numbers used in the analysis is available from the authors upon request. The tree was computed by using not cluster with rodent-associated MEGA5 (http://www.megasoftware.net). The Tamura 3-parameter model with gamma- hantaviruses but groups with those distributed rate heterogeneity and a proportion of invariant sites (T92 +G + I) was selected found in shrews and moles (Figure). as the best fi t evolutionary model according to the Baeysian information criterion calculated Considering that bats are with MEGA5. The values at the tree branches are the bootstrap support values calculated more closely related to shrews and from 500 replicates. Scale bar indicates an evolutionary distance of 0.2 substitutions per position in the sequence. The gray areas indicate association of hantaviruses with moles than to rodents (6), a certain reservoir host families. The MGB/1209 partial sequence of the large genomic segment was genetic similarity of a putative deposited in GenBank under accession no. JN037851.

160 Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 18, No. 1, January 2012 LETTERS the genera Eptesicus and Rhinolophus Sabrina Weiss,1 6. Murphy WJ, Eizirik E, Johnson WE, in South Korea. However, nucleotide Peter T. Witkowski,1 Zhang YP, Ryder OA, O’Brien SJ. Molec- ular phylogenetics and the origins of pla- sequencing showed the presence of Brita Auste, Kathrin Nowak, cental mammals. Nature. 2001;409:614– prototypical Hantaan virus indicating Natalie Weber, Jakob Fahr, 8. doi:10.1038/35054550 a spillover infection or laboratory Jean-Vivien Mombouli, 7. Jung YT, Kim GR. Genomic characteriza- contamination (7). Nathan D. Wolfe, tion of M and S RNA segments of hanta- viruses isolated from bats. Acta Virol. Further screening is necessary Jan Felix Drexler, 1995;39:231–3. to confi rm N. hispida as a natural Christian Drosten, 8. Schönrich G, Rang A, Lutteke N, Raf- reservoir host of the virus. Although Boris Klempa, tery MJ, Charbonnel N, Ulrich RG. the presented bat-associated Fabian H. Leendertz, Hantavirus-induced immunity in rodent reservoirs and humans. Immunol Rev. sequence is obviously distinct from and Detlev H. Kruger 2008;225:163–89. doi:10.1111/j.1600- other hantaviruses, which suggests Author affi liations: Robert Koch-Institute, 065X.2008.00694.x association with a novel natural host, Berlin, Germany (S. Weiss, K. Nowak, F.H. 9. Coulaud X, Chouaib E, Georges AJ, Rol- lin P, Gonzalez JP. First human case of a spillover infection from another, Leendertz); Charité School of Medicine, haemorrhagic fever with renal syndrome yet unrecognized host cannot be Berlin (P.T. Witkowski, B. Auste, B. Klempa, in the Central African Republic. Trans ruled out. However, detection of the D.H. Kruger); Ulm University, Ulm, Germany R Soc Trop Med Hyg. 1987;81:686. virus exclusively in 1 organ (lung (N. Weber); University of Braunschweig, doi:10.1016/0035-9203(87)90455-X 10. Klempa B, Koivogui L, Sylla O, Koulemou but not in liver, kidney, and spleen; Braunschweig, Germany (J. Fahr); K, Auste B, Kruger DH, et al. Serologi- data not shown) suggests a persistent Laboratoire National de Santé Publique, cal evidence of human hantavirus infec- infection that is typically observed in Brazzaville, Republic of the Congo (J.-V. tions in Guinea, West Africa. J Infect Dis. natural hosts of hantaviruses (8). Mombouli); Stanford University Program in 2010;201:1031–4. doi:10.1086/651169 To date, only a few reports exist Human Biology, Stanford, California, USA on cases of hemorrhagic fever with (N.D. Wolfe); University of Bonn Medical Address for correspondence: Detlev H. Kruger, renal syndrome in Africa (9,10). Centre, Bonn, Germany (J.F. Drexler, C. Institute of Medical Virology, Helmut-Ruska- However, underreporting must be Drosten); and Slovak Academy of Sciences, Haus, Charité Medical School, Charitéplatz assumed because the symptoms Bratislava, Slovakia (B. Klempa) 1, D-10117 Berlin, Germany; email: detlev. resemble those of many other febrile [email protected] DOI: http://dx. doi.org/10.3201/eid1801.111026 infections. Moreover, in cases of infections by non–rodent-associated References hantaviruses, cross-reactivity with routinely used rodent-borne virus 1. Krüger DH, Schonrich G, Klempa B. antigens should be limited and may Human pathogenic hantaviruses and hamper human serodiagnostics prevention of infection. Hum Vaccin. 2011;7:685–93. doi:10.4161/hv.7.6.15197 (1). The results suggest that bats, 2. Klempa B, Fichet-Calvet E, Lecompte Outbreak of which are hosts of many emerging E, Auste B, Aniskin V, Meisel H, et al. pathogens (5), may act as natural Hantavirus in African wood mouse, Guin- Porcine Epidemic reservoirs for hantavirus. The effect ea. Emerg Infect Dis. 2006;12:838–40. 3. Klempa B, Fichet-Calvet E, Lecompte E, Diarrhea in of this virus on public health remains Auste B, Aniskin V, Meisel H, et al. Novel Suckling Piglets, to be determined. hantavirus sequences in shrew, Guin- ea. Emerg Infect Dis. 2007;13:520–2. China doi:10.3201/eid1303.061198 This study was supported by 4. Kang HJ, Kadjo B, Dubey S, Jacquet F, To the Editor: Beginning in Deutsche Forschungsgemeinschaft (grant Yanagihara R. Molecular evolution of October 2010, porcine epidemic Azagny virus, a newfound hantavirus har- diarrhea (PED), caused by a coronaviral KR1293/9-1). Work in Sierra Leone was bored by the West African pygmy shrew supported by the Gola Forest Project. (Crocidura obscurior) in Cote d’Ivoire. infection affecting pigs, emerged in National and local authorities in Sierra Virol J. 2011;8:373. doi:10.1186/1743- China in an outbreak characterized by Leone, Liberia, Mali, Senegal, and 422X-8-373 high mortality rates among suckling 5. Calisher CH, Childs JE, Field HE, Holmes piglets. The outbreak overwhelmed Republic of Congo kindly granted research KV, Schountz T. Bats: important reservoir and export permits. N.D.W. is supported hosts of emerging viruses. Clin Micro- >10 provinces in southern China, by an award from the National Institutes biol Rev. 2006;19:531–45. doi:10.1128/ and >1,000,000 piglets died. This of Health Director’s Pioneer Award (grant CMR.00017-06 outbreak was distinguished by ≈100% illness among piglets after birth DP1-OD000370). 1These authors contributed equally to this (predominantly within 7 days and article.

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