Fatal Monkeypox in Wild-Living Sooty Mangabey, Côte d’Ivoire, 2012

Aleksandar Radonić, Sonja Metzger, Piotr Wojtek Dabrowski, Emmanuel Couacy-Hymann, Livia Schuenadel, Andreas Kurth, Kerstin Mätz-Rensing, Christophe Boesch, Fabian H. Leendertz, and Andreas Nitsche

Author affiliations: Robert Koch Institute, Berlin, Germany (A. Radonić, S. Metzger, P.W. Dabrowski, L. Schuenadel, A. Kurth, F.H. Leendertz, A. Nitsche); Laboratoire de la Pathologie Animale, Bingerville, Côte d’Ivoire (E. Couacy-Hymann); Deutsches Primatenzentrum, Göttingen, Germany (K. Mätz-Rensing); Max-Planck-Institute for Evolutionary Anthropology, Leipzig, Germany (C. Boesch)

Abstract

We isolated a monkeypox virus from a wild-living monkey, a sooty mangabey, found dead in Taï National Park, Côte d’Ivoire, in March 2012. The whole-genome sequence obtained from this isolate and directly from clinical specimens showed its close relationship to monkeypox viruses from Western Africa.

Among the poxviruses are several species of orthopoxviruses (OPVs) that are pathogenic to humans, including monkeypox virus (MPXV) and variola virus (VARV). MPXV was first discovered in laboratory captive monkeys in Copenhagen in 1958 (1). After the eradication of VARV during the 1970s, MPXV became the highest pathogenic OPV infection in humans. On the basis of epidemiologic and sequence data, strains of MPXV can be assigned to a West African or a Congo Basin clade; viruses from the Congo Basin clade show more pronounced illness, death, viremia, and human-to-human transmission than do strains from the West African clade (2,3).

Serologic studies showed that monkeys from Africa have OPV antibodies, but no natural case of MPXV has been reported in wild-living primates (4,5). The only MPXV isolate obtained from an animal in the wild was from a Thomas’s rope squirrel (Funisciurus anerythrus) caught in Democratic Republic of the Congo in 1985 (6). Here we describe natural MPXV infection in a sooty mangabey (Cercocebus atys) found dead in Taï National Park (TNP), Côte d’Ivoire.

The Study

During a long-term program to monitor deaths in wildlife, an infant mangabey was found dead in the TNP in March 2012. The body did not show any apparent injuries, and the animal had died relatively recently, as indicated by the presence of blowfly eggs but absence of maggots. Multiple skin lesions typical of MPXV infection occurred as dark red crusts 5–7 mm in diameter, partly confluent, which were disseminated over the body. Extremities were mainly affected; fewer lesions were seen on the belly and none on the back.

A full necropsy was performed under high-level safety measures, and samples of all organs and blood were collected and preserved in liquid nitrogen and 10% buffered formalin (7). Histologic analysis of the skin showed eosinophilic inclusion bodies, suggesting that an OPV infection had caused the ulcers. Severe bacterial secondary infection of the ulcers also was observed, as well as bacteremia that might have contributed to the pathologic changes and death.

DNA was extracted from different tissues, and quantitative PCR for OPV DNA (rpo18) and a cellular target (c-myc) were performed as described (8). We found a high viral DNA load in relation to cellular DNA in all tissues, except muscle, indicating a systemic infection, with particularly high loads in a skin lesion and from a throat swab sample (Table). An immunofluorescence assay performed on MPXV-infected cells showed titers of 320 for IgG and 80 for IgM, indicating an acute OPV infection.

Virus from skin tissue was propagated in HEp-2 cells, infected cells were harvested, and DNA was extracted. We performed library preparation and sequencing on an Ion Torrent PGM with an Ion PGM Sequencing 200 Kit (Life Technologies, Darmstadt, Germany) (average read length = 93 bases). Sequences were analyzed by using Geneious (Biomatters, Auckland, NZ). At first, a de novo assembly from PGM fastq-data was conducted, followed by mapping of the resulting contigs to all 11 MPXV genomes published in the National Center for Biotechnology Information to obtain orientation and to form a consensus sequence. The reads were mapped again to the consensus sequences to identify assembly errors. These efforts resulted in a single 197,571-bp genome. Additionally, DNA preparations from the mangabey’s skin and throat swab specimens were subjected directly to an Illumina HiSeq 1500 (San Diego, CA, USA), sequencing 150 + 150 bases (paired end). The genome could also be assembled to 190,562 bp by using Illumina data only (2.9% of the 285 million Illumina reads could be used for the MPXV assembly, compared with 45% of the 3.5 million reads from the PGM), indicating that the viral genome can be assembled by sequencing specimens of high viral load directly without previous virus propagation.

Dr Radonić is a research scientist at the Robert Koch Institute. His primary research focus is the implementation, development, validation, and diagnostic applications of next-generation sequencing approaches.

Acknowledgment

We thank the Ivorian authorities for long-term support, especially the Ministry of the Environment and Forests, as well as the Ministry of Research, the directorship of the Taï National Park and the Swiss Research Center. We also thank Jule Hinzmann, Ines Müller, Jung-Won Sim-Brandenburg, and Delia Barz for their excellent technical assistance and Ursula Erikli for copy editing.

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Figures

Table

Table. Results of quantitative PCR from tissues of a wild-living sooty mangabey (Cercocebus atys), Taï National Park, Côte d’Ivoire, March 2012

Suggested citation for this article: Radonić A, Metzger S, Dabrowski PW, Couacy-Hymann E, Schuenadel L, Kurth A, et al. Fatal monkeypox in wild-living sooty mangabey, Côte d’Ivoire, 2012. Emerg Infect Dis [Internet]. 2014 Jun [date cited].http://dx.doi.org/10.3201/eid2006.131329