Skip to main content
SpringerLink
Log in

Prevalence and Phylogenetic Analysis of Hepatitis B in Captive and Wild-Living Pileated Gibbons (Hylobates pileatus) in Cambodia

  • Published:
International Journal of Primatology Aims and scope Submit manuscript

Abstract

The risk of transmitting novel pathogens from released animals to wild conspecifics is an important consideration for reintroduction initiatives. Apart from infecting humans, hepatitis B virus (HBV) has been detected in nonhuman primates, including captive gibbons. Gibbons that test positive for HBV are removed from reintroduction programs as the presence of infection in wild populations is uncertain. The aim of this study was to determine the prevalence of HBV in wild-living gibbon populations in the southwest and northeast of Cambodia and compare HBV DNA strains collected from these animals with samples from wild-born captive gibbons at the Phnom Tamao Wildlife Rescue Center, Cambodia. We screened HBV infection in blood and stool samples from captive gibbons and stool samples from wild-living gibbons. We sequenced samples with detectable HBV DNA viral load in the HBV reverse transcriptase coding region overlapping with the S-gene. Eleven of twenty-four (46 %) wild-living pileated gibbons (Hylobates pileatus) from the southwest of Cambodia were positive for HBV. Phylogenetic analysis of HBV DNA sequenced from wild and captive gibbons revealed that viral strains were related and different from human strains. The presence of an HBV variant circulating in the wild similar to that found in captive pileated gibbons suggests 1) the strain identified is associated with the species, 2) wild-born captive gibbons positive for this strain became infected prior to captivity, and 3) it might be appropriate to release captive gibbons that test positive for specific serological markers of HBV, provided they are otherwise suitable candidates for reintroduction.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Aiba, N., Nishimura, H., Arakawa, Y., & Abe, K. (2003). Complete nucleotide sequence and phylogenetic analyses of hepatitis B virus isolated from two pileated gibbons. Virus Genes, 27(3), 219–226. https://doi.org/10.1023/A:1026387614162.

    Article  CAS  PubMed  Google Scholar 

  • Ballou, J. D. (1993). Assessing the risks of infectious diseases in captive breeding and reintroduction programs. Journal of Zoo and Wildlife Medicine, 24(3), 327–335.

    Google Scholar 

  • Bancroft, W. H., Snitbhan, R., McNair Scott, R., Tingpalapong, M., Watson, W. T., et al (1977). Transmission of hepatitis B virus to gibbon by exposure to human saliva containing hepatitis B surface antigen. The Journal of Infectious Diseases, 135(1), 79–86.

    Article  CAS  PubMed  Google Scholar 

  • Beck, B., Cooper, M., & Griffith, B. (1993). Infectious disease considerations in reintroduction programs for captive wildlife. Journal of Zoo and Wildlife Medicine, 24(3), 394–397.

    Google Scholar 

  • Brockelman, W., Geissmann, T., Timmins, T., & Traeholt, C. (2008). Hylobates pileatus. The IUCN Red List of Threatened Species. Retrieved from http://www.iucnredlist.org. Accessed 09 Oct 2019.

  • Campbell, C. O., Cheyne, S. M., & Rawson, B. M. (2015). Best practice guidelines for the rehabilitation and translocation of gibbons. Gland, Switzerland: IUCN. https://doi.org/10.2305/iucn.ch.2015.ssc-op.51.en.

    Book  Google Scholar 

  • Cheyne, S. M. (2009). The role of reintroduction in gibbon conservation: Opportunities and challenges. In S. Lappan & D. J. Whittaker (Eds.), The gibbons (pp. 477–496). New York: Springer Science. https://doi.org/10.1007/978-0-387-88604-6.

    Chapter  Google Scholar 

  • Cheyne, S. M., Campbell, C. O., & Payne, K. L. (2012). Proposed guidelines for in situ gibbon rescue , rehabilitation and reintroduction. International Zoo Yearbook, 46, 1–17. https://doi.org/10.1111/j.1748-1090.2011.00149.x.

    Article  Google Scholar 

  • Cunningham, A. A. (1996). Disease risks of wildlife translocations. Conservation Biology, 10(2), 349–353.

    Article  Google Scholar 

  • Dominguez Souza, F. B. D. C., König, A., Rasche, A., Netto, E. M., Glebe, D., & Drexler, J. F. (2018). A novel hepatitis B virus species discovered in capuchin monkeys sheds new light on the evolution of primate hepadnaviruses. Journal of Hepatology, 68(6), 1114–1122. https://doi.org/10.1016/j.jhep.2018.01.029.

    Article  CAS  Google Scholar 

  • Geissmann, T., Manh Ha, N., Rawson, B., Timmins, R., Traeholt, C. & Walston, J. (2008). Nomascus gabriellae. The IUCN Red List of Threatened Species. Retrieved from http://www.iucnredlist.org. Accessed 09 Oct 2019.

  • Gomes-Gouvea, M. S., Soares, M. C. P., Bensabath, G., de Carvalho-Mello, I. M. V. G., Brito, E. M. F., et al (2009). Hepatitis B virus and hepatitis delta virus genotypes in outbreaks of fulminant hepatitis (Labrea black fever ) in the western Brazilian Amazon region. Journal of General Virology, 90, 2638–2643. https://doi.org/10.1099/vir.0.013615-0.

    Article  CAS  PubMed  Google Scholar 

  • Grethe, S., Heckel, J.-O., Rietschel, W., & Hufert, F. T. (2000). Molecular epidemiology of hepatitis B virus variants in nonhuman primates. Journal of Virology, 74(11), 5377–5381. https://doi.org/10.1128/jvi.74.11.5377-5381.2000.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Heckel, J.-O., Rietschel, W., & Hufert, F. T. (2001). Prevalence of hepatitis B virus infections in nonhuman primates. Journal of Medical Primatology, 30, 14–19. https://doi.org/10.1128/jvi.74.11.5377-5381.2000.

    Article  CAS  PubMed  Google Scholar 

  • Huang, C.-C., Chiang, Y.-C., Chang, C.-D., & Wu, Y.-H. (2009). Prevalence and phylogenetic analysis of hepatitis B virus among nonhuman primates in Taiwan. Journal of Zoo and Wildlife Medicine, 40(3), 519–528. https://doi.org/10.1638/2008-0150.1.

    Article  CAS  PubMed  Google Scholar 

  • IUCN/SCC (1998). Guidelines for re-introductions. Gland, Switzerland and Cambridge, UK: IUCN.

    Google Scholar 

  • IUCN/SCC (2013). Guidelines for reintroductions and other conservation translocations. Version 1.0. Gland, Switzerland: IUCN.

    Google Scholar 

  • Kania, D., Ottomani, L., Meda, N., Peries, M., Dujols, P., et al (2014). Performance of two real-time PCR assays for hepatitis B virus DNA detection and quantitation. Journal of Virological Methods, 201, 24–30. https://doi.org/10.1016/j.jviromet.2014.01.015.

    Article  CAS  PubMed  Google Scholar 

  • Kock, R., Woodford, M. H., & Rossiter, P. B. (2010). Disease risks associated with the translocation of wildlife. Scientific and Technical Review of the Office International des Epizooties, 29(2), 329–350. https://doi.org/10.20506/rst.29.2.1980.

    Article  CAS  Google Scholar 

  • Kumar, S., Stecher, G., & Tamura, K. (2016). MEGA7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets. Society for Molecular Biology and Evolution, 1–11.

  • Kumar, S., Stecher, G., Li, M., Knyaz, C., & Tamura, K. (2018). MEGA X: Molecular evolutionary genetics analysis across computing platforms. Molecular Biology and Evolution, 35(6), 1547–1549. https://doi.org/10.1093/molbev/msy096.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lanford, R. E., Chavez, D., Brasky, K. M., Burns, R. B., & Rico-Hesse, R. (1998). Isolation of a hepadnavirus from the woolly monkey, a New World primate. Proceedings of the National Academy of Sciences of the USA, 95(10), 5757–5761. https://doi.org/10.1073/pnas.95.10.5757.

    Article  CAS  PubMed  Google Scholar 

  • Lanford, R. E., Chavez, D., Rico-Hesse, R., & Mootnick, A. (2000). Hepadnavirus infection in captive gibbons. Journal of Virology, 74(6), 2955–2959. https://doi.org/10.1128/jvi.74.6.2955-2959.2000.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lyons, S., Sharp, C., LeBreton, M., Djoko, C. F., Kiyang, J. A., et al (2012). Species association of hepatitis b virus (HBV) in non-human apes; Evidence for recombination between gorilla and chimpanzee variants. PLoS ONE, 7(3), 1–9. https://doi.org/10.1371/journal.pone.0033430.

    Article  CAS  Google Scholar 

  • MacDonald, D. M., Holmes, E. C., Lewis, J. C. M., & Simmonds, P. (2000). Detection ofhepatitis B virus infection in wild-born chimpanzees (Pan troglodytes verus): Phylogenetic relationships with human and other primate genotypes. Journal of Virology, 74(9), 4253–4257. https://doi.org/10.1128/jvi.74.9.4253-4257.2000.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Makuwa, M., Souquière, S., Telfer, P., Leroy, E., Bourry, O., et al (2003). Occurrence of hepatitis viruses in wild-born non-human primates: A 3 year (1998–2001) epidemiological survey in Gabon. Journal of Medical Primatology, 32, 307–314.

    Article  CAS  PubMed  Google Scholar 

  • Makuwa, M., Souquière, S., Clifford, S. L., Mouinga-Ondeme, A., Bawe-Johnson, M., et al (2005). Identification of hepatitis B virus genome in faecal sample from wild living chimpanzee (Pan troglodytes troglodytes) in Gabon. Journal of Clinical Virology, 34(1), 83–88. https://doi.org/10.1016/S1386-6532(05)80016-2.

    Article  CAS  Google Scholar 

  • Makuwa, M., Souquière, S., Bourry, O., Rouquet, P., Telfer, P., et al (2007). Complete-genome analysis of hepatitis B virus from wild-born chimpanzees in central Africa demonstrates a strain-specific geographical cluster. Journal of General Virology, 88, 2679–2685. https://doi.org/10.1099/vir.0.82992-0.

    Article  CAS  PubMed  Google Scholar 

  • Mathews, F., Moro, D., Strachan, R., Gelling, M., & Buller, N. (2006). Health surveillance in wildlife reintroductions. Biological Conservation, 131(2), 338–347. https://doi.org/10.1016/j.biocon.2006.04.011.

    Article  Google Scholar 

  • Mimms, L. T., Solomon, L. R., Ebert, J. W., & Fields, H. (1993). Unique pre-S sequence in a gibbon-derived hepatitis B virus variant. Biochemical and Biophysical Research Communications, 195(1), 186–191.

    Article  CAS  PubMed  Google Scholar 

  • Noppornpanth, S., Haagmans, B. L., Bhattarakosol, P., Ratanakorn, P., Niesters, H. G. M., et al (2003). Molecular epidemiology of gibbon hepatitis B virus transmision. Journal of General Virology, 84(1), 147–155. https://doi.org/10.1099/vir.0.18531-0.

    Article  CAS  PubMed  Google Scholar 

  • Norder, H., Ebert, J. W., Fields, H. A., Mushahwar, I. K., & Magnius, L. O. (1996). Complete sequencing of a gibbon hepatitis B virus genome reveals a unique genotype distantly related to the chimpanzee hepatitis B virus. Virology, 218(1), 214–223. https://doi.org/10.1006/viro.1996.0181.

    Article  CAS  PubMed  Google Scholar 

  • Payne, K. L., Monaghan, C. L., Warren, K. S., Fenwick, S., Robertson, I., & Smith, D. (2003). Hepatitis B virus vaccination of silvery gibbons (Hylobates moloch) at the Perth Zoo. In 2003 Proceedings of the American Association of Zoo Veterinarians (pp. 187–191).

  • Riley, E. P., Mackinnon, K. C., Fernandez-Duque, E., Setchell, J. M., & Garber, P. A. (2014). Code of best practices for field primatology. International Primatological Society and American Society of Primatologists Steering Committee. doi:https://doi.org/10.13140/2.1.2889.1847

  • Roos, C., Booratana, R., Supriatna, J., Fellowes, J. R., Rylands, A. B., & Mittermeier, R. A. (2013). An updated taxonomy of primates in Vietnam, Laos, Cambodia and China. Vietnamese Journal of Primatology, 2(2), 13–26.

    Google Scholar 

  • Sall, A. A., Starkman, S., Reynes, J. M., Lay, S., Nhim, T., et al (2005). Frequent infection of Hylobates pileatus (pileated gibbon) with species-associated variants of hepatitis B virus in Cambodia. Journal of General Virology, 86(2), 333–337. https://doi.org/10.1099/vir.0.80274-0.

    Article  CAS  PubMed  Google Scholar 

  • Scott, R. M., Snitbhan, R., Bancroft, W. H., Alter, H. J., & Tingpalapong, M. (1980). Experimental transmission of hepatitis B virus by semen and saliva. The Journal of Infectious Diseases, 142(1), 67–71.

    Article  CAS  PubMed  Google Scholar 

  • Sitnik, R., Pinho, J. R. R., Bertolini, D. A., Bernardini, A. P., da Silva, L. C., & Carrilho, F. J. (2004). Hepatitis B virus genotypes and precore and core mutants in Brazilian patients. Journal of Clinical Microbiology, 42(6), 2455–2460. https://doi.org/10.1128/JCM.42.6.2455.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Starkman, S. E., MacDonald, D. M., Lewis, J. C. M., Holmes, E. C., & Simmonds, P. (2003). Geographic and species association of hepatitis B virus genotypes in non-human primates. Virology, 314, 381–393. https://doi.org/10.1016/S0042-6822(03)00430-6.

    Article  CAS  PubMed  Google Scholar 

  • Thinh, V. N., Mootnick, A. R., Geissmann, T., Li, M., Ziegler, T., et al (2010a). Mitochondrial evidence for multiple radiations in the evolutionary history of small apes. BMC Evolutionary Biology, 10(1), 74.

    Article  PubMed  PubMed Central  Google Scholar 

  • Thinh, V. N., Mootnick, A. R., Thanh, V. N., Nadler, T., & Roos, C. (2010b). A new species of crested gibbon, from the central Annamite mountain range. Vietnamese Journal of Primatology, 4, 1–12.

    Google Scholar 

  • Thornton, S. M., Walker, S., & Zuckerman, J. N. (2001). Management of hepatitis B virus infections in two gibbons and a western lowland gorilla in a zoological collection. Veterinary Record, 149, 113–115.

    Article  CAS  PubMed  Google Scholar 

  • Traeholt, C., Bunthoeun, R., Rawson, B., Samuth, M., Virak, C., & Vuthin, S. (2005). Status review of pileated gibbon, hylobates pileatus, and yellow-cheeked crested gibbon, Nomascus gabriellae, Cambodia. Phnom Penh: Fauna & Flora International – Indochina Programme.

    Google Scholar 

  • Travis, D. A., Hungerford, L., Engel, G. A., & Jones-Engel, L. (2006). Diseases risk analysis: A tool for primate conservation planning and decision making. American Journal of Primatology, 68, 855–867. https://doi.org/10.1002/ajp.

    Article  CAS  PubMed  Google Scholar 

  • Viggers, K. L., Lindenmayer, D. B., & Spratt, D. M. (1993). The importance of disease in reintroduction programmes. Wildlife Research, 20(5), 687–698. https://doi.org/10.1071/WR9930687.

    Article  Google Scholar 

  • Warren, K. S., Heeney, J. L., Swan, R. A., & Verschoor, E. J. (1999). A new group of hepadnaviruses naturally infecting orangutans (Pongo pygmaeus). Journal of Virology, 73(9), 7860–7865 http://jvi.asm.org/.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Woodford, M. H. (2000). Quarantine and health screening protocols for wildlife prior to translocation and release into the wild. Other Publications in Zoonotics and Wildlife Disease, 32, 7–27.

    Google Scholar 

  • Zheng, J. S., Chen, M. M., Yang, H. F., Zhou, X. T., Liu, Y. Y., Li, J., & Bin (2015). Stool vs. serum hepatitis B virus DNA in patients with chronic hepatitis B. Medical Science Monitor, 21, 3840–3846. https://doi.org/10.12659/MSM.896243.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

The authors would like to thank the following organizations and individuals for their support and assistance in this research: Wildlife Reserves Singapore (WRS) for providing the total funding in both years of research into this important study; Dr Sonja Luz for her support and particularly for her understanding when confronted with the unexpected; the Aspinall Foundation, which conducted a similar research and whose support and advice contributed greatly to this study; Dr. Jane Hopper, whose unpublished paper on HBV in wild gibbons, implemented by the Aspinall Foundation, gave guidance; Mr. Ernie Thetford, for his advice and expertise on all things primate; Mr. Nhek Ratanapich, Director of Phnom Tamao Wildlife Rescue Centre, for his agreement and support for this and many other projects concerning the welfare of his animals; Wildlife Alliance staff involved at Phnom Tamao Wildlife Rescue Centre and at our Wildlife Release Station in Koh Kong province, who would otherwise have gone unnoticed, particularly vets Oung Chenda, Ruos San and Oung Chamroeun and head keeper Try Sitheng at PTWRC and Chhean Soeun, Toeur Savat and Kong Vang in Koh Kong; and the Cambodian Forestry Administration, Ministry of Agriculture Forestry and Fisheries for its support. We also thank the editor and reviewers for all their thoughtful comments and assistance with improving our report.

Author information

Author notes

  1. Nicole Leroux and Janin Nouhin contributed equally to this work.

Authors and Affiliations

  1. Wildlife Rescue and Care Programs, Wildlife Alliance, Phnom Penh, Cambodia

    Nicole Leroux, Bunthoeun Roth & Nick Marx

  2. Virology Unit, Institut Pasteur du Cambodge, Institut Pasteur International Network, Phnom Penh, Cambodia

    Janin Nouhin, Sophearot Prak, François Rouet & Philippe Dussart

  3. Institute of Forest and Wildlife Research, Forestry Administration, Phnom Penh, Cambodia

    Bunthoeun Roth

Author notes

  1. François Rouet is deceased. This paper is dedicated to his memory.

    • François Rouet
Authors
  1. Nicole Leroux
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Janin Nouhin
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sophearot Prak
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Bunthoeun Roth
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. François Rouet
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Philippe Dussart
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Nick Marx
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

NM and RB formulated the idea. NM, JN, and RB developed methodology and experiment design. NL, RB, and NM organized and oversaw field work to collect samples. JN, SP, FR, and PD developed methodology for and preformed laboratory analysis of samples. NL, JN, PD and NM wrote the manuscript and the other authors provided input and editorial advice.

Corresponding author

Correspondence to Nicole Leroux.

Additional information

Handling Editor: Joanna M. Setchell

Electronic supplementary material

ESM 1

(DOCX 21.4 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Leroux, N., Nouhin, J., Prak, S. et al. Prevalence and Phylogenetic Analysis of Hepatitis B in Captive and Wild-Living Pileated Gibbons (Hylobates pileatus) in Cambodia. Int J Primatol 41, 634–653 (2020). https://doi.org/10.1007/s10764-020-00168-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10764-020-00168-1

Keywords

Search

Navigation