Nyctinomops laticaudatus bat-associated Rabies virus causes disease with a shorter clinical period and has lower pathogenic potential than strains isolated from wild canids

  • Natalia Langenfeld Fuoco
  • Elaine Raniero Fernandes
  • Fernanda Guedes
  • Sandriana Dos Ramos Silva
  • Leticia Patricia Guimarães
  • Nayara Ugeda Silva
  • Orlando Garcia Ribeiro
  • Iana Suly Santos KatzEmail author
Original Article


Rabies is a lethal viral disease that can affect a wide range of mammals. Currently, Rabies virus (RABV) in some European and American countries is maintained primarily in wild species. The regulation of viral replication is one of the critical mechanisms involved in RABV pathogenesis. However, the relationship between replication and the pathogenesis of RABV isolated from wild animals remains poorly understood. In the present study, we evaluated the pathogenicity of the street viruses Nyctinomops laticaudatus bat-associated RABV (NYBRV) and Cerdocyon thous canid-associated RABV (CECRV). Infection of mice with NYBRV led to 33% mortality with rapid disease evolution and marked histopathological changes in the CNS. In contrast, infection with CECRV led to 67% mortality and caused mild neuropathological lesions. The proportion of RABV antigen was significantly higher in the cytoplasm of neuronal cells of the cerebral cortex and in the meninges of mice infected with CECRV and NYBRV, respectively. Moreover, the replication rate of NYBRV was significantly higher (p < 0.001) than that of CECRV in neuroblastoma cells. However, CECRV replicated to a significantly higher titer in epithelial cells. Our results indicate that NYBRV infection results in rapid disease progression accompanied by frequent and intense histopathological alterations in the CNS in mice, and in a high replication rate in neuroblastoma cells. Although, CECRV is more pathogenic in mice, it caused milder histopathological changes in the CNS and replicated more efficiently in epithelial cells. Our data point to a correlation between clinical aspects of disease and the replication of RABV in different cell lines.



We would like to thank Karen Myiuki Asano at the Pasteur Institute for providing the mice. This work was supported by Instituto Pasteur, São Paulo, Brazil.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

Animal rights statement

The study protocol was approved by the Committee on the Ethics of Animal Experiments of the Instituto Pasteur of São Paulo. All institutional and national guidelines for the care and use of laboratory animals were followed.

Consent for publication

All authors consent to publication.


  1. 1.
    Rupprecht CE, Hanlon CA, Hemachudha T (2002) Rabies re-examined. Lancet Infect Dis 2:327–343CrossRefGoogle Scholar
  2. 2.
    Favoretto SR, de Mattos CC, de Mattos CA et al (2013) The emergence of wildlife species as a source of human rabies infection in Brazil. Epidemiol Infect 141:1552–1561. CrossRefGoogle Scholar
  3. 3.
    Messenger SL, Smith JS, Orciari LA et al (2003) Emerging pattern of rabies deaths and increased viral infectivity. Emerg Infect Dis 9(2):151–4. CrossRefGoogle Scholar
  4. 4.
    Escobar LE, Peterson T, Favi M et al (2015) Bat-borne rabies in latin America. Rev Inst Med Trop Sao Paulo 57:63–72. CrossRefGoogle Scholar
  5. 5.
  6. 6.
    Rocha SM, de Oliveira SV, Heinemann MB, Gonçalves VSP (2017) Epidemiological profile of wild rabies in Brazil (2002–2012). Transbound Emerg Dis 64:624–633. CrossRefGoogle Scholar
  7. 7.
    Castilho JG, Achkar SM, de Novaes Oliveira R et al (2018) Analysis of rabies diagnosis in dogs and cats in the state of São Paulo, Brazil. Arch Virol 163:2369–2376. CrossRefGoogle Scholar
  8. 8.
    Appolinário C, Dora S, Ferreira A et al (2015) Fluorescent antibody test, quantitative PCR pattern and clinical aspects of rabies virus strains isolated from main reservoirs in Brazil. Braz J Infect Dis 19(5):479–85. CrossRefGoogle Scholar
  9. 9.
    Cunha EMS, Nassar AFC, Lara MCCSH et al (2010) Pathogenicity of different rabies virus isolates and protection test in vaccinated mice. Rev Inst Med Trop Sao Paulo 52(5):231–236. CrossRefGoogle Scholar
  10. 10.
    Katz ISS, Fuoco NL, Chaves LB et al (2016) Delayed progression of rabies transmitted by a vampire bat. Arch Virol 161:2561–2566. CrossRefGoogle Scholar
  11. 11.
    Fuoco NL, Fernandes ER, Dos Ramos Silva S, Luiz FG, Ribeiro OGSKI (2018) Street rabies virus strains associated with insectivorous bats are less pathogenic than strains isolated from other reservoirs. Antiviral Res 160:94–100. CrossRefGoogle Scholar
  12. 12.
    Dietzschold B, Li J, Faber M, Schnell M (2008) Concepts in the pathogenesis of rabies. Future Virol 3:481–490. CrossRefGoogle Scholar
  13. 13.
    Faber M, Faber M-L, Li J et al (2007) Dominance of a nonpathogenic glycoprotein gene over a pathogenic glycoprotein gene in rabies virus. J Virol 81:7041–7047. CrossRefGoogle Scholar
  14. 14.
    Wang X, Zhang S, Sun C et al (2011) Proteomic profiles of mouse neuro N2a cells infected with variant virulence of rabies viruses. J Microbiol Biotechnol 21:366–373. CrossRefGoogle Scholar
  15. 15.
    Feng YJ (2014) Original article comparative analysis of the pathogenic mechanisms of street rabies virus strains with different virulence levels. Biomed Environ Sci 27:749–762. Google Scholar
  16. 16.
    Castilho JG, Iamamoto K, Yoshitaka J, et al (2007) Padronização e aplicação da técnica de isolamento do vírus da raiva em células de neuroblastoma de camundongo (N2A). Bol Epidemiológico Paul (BEPA) 4:12–18Google Scholar
  17. 17.
    Fuoco NL, dos Ramos Silva S, Fernandes ER et al (2018) Infection of neuroblastoma cells by rabies virus is modulated by the virus titer. Antiviral Res 149:89–94. CrossRefGoogle Scholar
  18. 18.
    Yamada K, Park CH, Noguchi K et al (2012) Serial passage of a street rabies virus in mouse neuroblastoma cells resulted in attenuation: potential role of the additional N-glycosylation of a viral glycoprotein in the reduced pathogenicity of street rabies virus. Virus Res 165:34–45. CrossRefGoogle Scholar
  19. 19.
    Wild TF, Bijlenga G (1981) A rabies virus persistent infection in BHK21 cells. J Gen Virol 57:169–177. CrossRefGoogle Scholar
  20. 20.
    Zhu S, Guo C (2016) Rabies control and treatment: From prophylaxis to strategies with curative potential. Viruses 8:1–23. CrossRefGoogle Scholar
  21. 21.
    Faber M, Faber M-L, Li J et al (2007) Dominance of a nonpathogenic glycoprotein gene over a pathogenic glycoprotein gene in rabies virus. J Virol. Google Scholar
  22. 22.
    Faber M, Faber M, Papaneri A et al (2005) A single amino acid change in rabies virus glycoprotein increases virus spread and enhances virus pathogenicity a single amino acid change in rabies virus glycoprotein increases virus spread and enhances virus pathogenicity. J Virol 79:14141–14148. CrossRefGoogle Scholar
  23. 23.
    Assis and Rosemberg (1984) Human rabies. Neuropathological study of thirty cases. Rev Inst Med Trop Sao Paulo 26:346–352CrossRefGoogle Scholar
  24. 24.
    Fernandes ER, de Andrade HF, Lancellotti CLP et al (2011) In situ apoptosis of adaptive immune cells and the cellular escape of rabies virus in CNS from patients with human rabies transmitted by Desmodus rotundus. Virus Res 156:121–126. CrossRefGoogle Scholar
  25. 25.
    Nogueira YL (2004) Estimate of the validity of a new method for the isolation of rabies virus. Rev Saude Publica 38:315–322CrossRefGoogle Scholar
  26. 26.
    Favoretto SR, De Mattos CC, De Morais NB et al (2006) Rabies virus maintained by dogs in humans and terrestrial wildlife, Ceará State, Brazil. Emerg Infect Dis 12:1978–1981. CrossRefGoogle Scholar
  27. 27.
    Preuss MAR, Faber ML, Tan GS et al (2009) Intravenous inoculation of a bat-associated rabies virus causes lethal encephalopathy in mice through invasion of the brain via neurosecretory hypothalamic fibers. PLoS Pathog 5(6):e1000485. CrossRefGoogle Scholar
  28. 28.
    Jackson AC (2003) Rabies virus infection: an update. J Neurovirol 9:253–258. CrossRefGoogle Scholar
  29. 29.
    Jackson AC (2010) Rabies pathogenesis update. Rev Pan-Amazônica Saúde 1:167–172. Google Scholar
  30. 30.
    Carnieli P, Ruthner Batista HBC, de Novaes Oliveira R et al (2013) Phylogeographic dispersion and diversification of rabies virus lineages associated with dogs and crab-eating foxes (Cerdocyon thous) in Brazil. Arch Virol 158:2307–2313. CrossRefGoogle Scholar
  31. 31.
    Carnieli P, Fahl WDO, Castilho JG et al (2008) Characterization of Rabies virus isolated from canids and identification of the main wild canid host in Northeastern Brazil. Virus Res 131:33–46. CrossRefGoogle Scholar
  32. 32.
    Carnieli Jr P, Scheffer KC, Chaves LB et al (2011) Dogs as reservoir and transmitter of the rabies virus.Vincent M. DeGiovine. (Org.). Dogs: Biology, Behavior and Health Disorders. Nova Science Publishers, Inc., New YorkGoogle Scholar
  33. 33.
    Carnieli P, Castilho JG, Fahl WDO et al (2009) Molecular characterization of Rabies Virus isolates from dogs and crab-eating foxes in Northeastern Brazil. Virus Res 141:81–89. CrossRefGoogle Scholar
  34. 34.
    Oliveira RDN, de Souza SP, Lobo RSV et al (2010) Rabies virus in insectivorous bats: Implications of the diversity of the nucleoprotein and glycoprotein genes for molecular epidemiology. Virology 405:352–360. CrossRefGoogle Scholar
  35. 35.
    Castilho JG, Carnieli P, Oliveira RN et al (2010) A comparative study of rabies virus isolates from hematophagous bats in Brazil. J Wildl Dis 46:1335–1339. CrossRefGoogle Scholar
  36. 36.
    Castilho JG, Canello FM, Scheffer KC et al (2008) Antigenic and genetic characterization of the first rabies virus isolated from the bat Eumops perotis in Brazil. Rev Inst Med Trop Sao Paulo 50:95–99. CrossRefGoogle Scholar
  37. 37.
    Menozzi BD, de Novaes Oliveira R, Paiz LM et al (2017) Antigenic and genotypic characterization of rabies virus isolated from bats (Mammalia: Chiroptera) from municipalities in São Paulo State, Southeastern Brazil. Arch Virol 162:1201–1209. CrossRefGoogle Scholar
  38. 38.
    de Souza DN, Carnieli P, Macedo CI et al (2017) Phylogenetic analysis of rabies virus isolated from canids in North and Northeast Brazil. Arch Virol 162:71–77. CrossRefGoogle Scholar
  39. 39.
    Charlton KM, Nadin-Davis S, Casey GA, Wandeler AI (1997) The long incubation period in rabies: Delayed progression of infection in muscle at the site of exposure. Acta Neuropathol 94:73–77. CrossRefGoogle Scholar
  40. 40.
    Healy DM, Brookes SM, Banyard AC et al (2013) Pathobiology of rabies virus and the European bat lyssaviruses in experimentally infected mice. Virus Res 172(1–2):46–53. CrossRefGoogle Scholar
  41. 41.
    Baer GM, Cleary WF, Diaz AM, Perl DF (1977) Characteristics of 11 rabies virus isolates in mice: titers and relative invasiveness of virus, incubation period of infection, and survival of mice with sequelae. J Infect Dis 136:336–345. CrossRefGoogle Scholar
  42. 42.
    Garcia SA, Lebrun A, Kean RB, Craig Hooper D (2018) Clearance of attenuated rabies virus from brain tissues is required for long-term protection against CNS challenge with a pathogenic variant. J Neurovirol 24:606–615. CrossRefGoogle Scholar
  43. 43.
    Roy A, Phares TW, Koprowski H, Hooper DC (2006) Failure to open the blood-brain barrier and deliver immune effectors to central nervous system tissues leads to the lethal outcome of silver-haired bat rabies virus infection. J Virol 81:1110–1118. CrossRefGoogle Scholar
  44. 44.
    Wang ZW, Sarmento L, Wang Y, Li XQ, Dhingra V, Tseggai T, Jiang BFZ (2005) Attenuated rabies virus activates, while pathogenic rabies virus evades, the host innate immune responses in the central nervous system. J Virol 79:12554–12565. CrossRefGoogle Scholar
  45. 45.
    Brzozka K, Finke S, Conzelmann K-K (2006) Inhibition of interferon signaling by rabies virus phosphoprotein P: activation-dependent binding of STAT1 and STAT2. J Virol 80:2675–2683. CrossRefGoogle Scholar
  46. 46.
    Luco S, Delmas O, Vidalain PO et al (2012) RelAp43, a member of the NF-κB family involved in innate immune response against lyssavirus infection. PLoS Pathog 8(12):e1003060. CrossRefGoogle Scholar
  47. 47.
    Masatani T, Ito N, Shimizu K, Ito Y, Nakagawa K, Sawaki Y, Koyama HSM (2010) Rabies virus nucleoprotein functions to evade activation of the RIG-I-mediated antiviral response. J Virol 84:4002–4012. CrossRefGoogle Scholar
  48. 48.
    Kopitar-Jerala N (2017) The role of interferons in inflammation and inflammasome activation. Front Immunol 8:873. CrossRefGoogle Scholar
  49. 49.
    Jackson AC (2016) Diabolical effects of rabies encephalitis. J Neurovirol 22:8–13. CrossRefGoogle Scholar
  50. 50.
    Katz ISS, Guedes F, Fernandes ER, dos Ramos Silva S (2017) Immunological aspects of rabies: a literature review. Arch Virol 162:3251–3268. CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  • Natalia Langenfeld Fuoco
    • 1
  • Elaine Raniero Fernandes
    • 1
  • Fernanda Guedes
    • 1
  • Sandriana Dos Ramos Silva
    • 1
  • Leticia Patricia Guimarães
    • 1
  • Nayara Ugeda Silva
    • 1
  • Orlando Garcia Ribeiro
    • 2
  • Iana Suly Santos Katz
    • 1
    Email author
  1. 1.Pasteur InstituteSão PauloBrazil
  2. 2.Laboratory of ImmunogeneticsButantan InstituteSão PauloBrazil

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