In 2007, we detected human herpes simplex virus type 1, which caused stomatitis, in a juvenile confiscated eastern lowland gorilla (Gorilla beringei graueri) that had a high degree of direct contact with human caretakers. Our findings confirm that pathogens can transfer between nonhuman primate hosts and humans

Barry, Peter A

Cranfield, Michael R

Gardner-Roberts, David

Gilardi, Kirsten VK

Kinani, Jean-Felix

Lowenstine, Linda J

Oxford, Kristie L

Spelman, Lucy

eScholarship - University of California

UC DavisUC Davis Previously Published WorksTitleHuman Herpes Simplex Virus Type 1 in Confiscated Gorilla - Volume 20, Number 11—November 2014 - Emerging Infectious Diseases journal - CDCPermalinkhttps://escholarship.org/uc/item/2n56p06dJournalEmerging Infectious Diseases, 20(11)ISSN1080-6040AuthorsGilardi, Kirsten VKOxford, Kristie LGardner-Roberts, Davidet al.Publication Date2014-11-01DOI10.3201/eid2011.140075 Peer reviewedeScholarship.org Powered by the California Digital LibraryUniversity of CaliforniaHuman Herpes Simplex Virus Type 1 in Confiscated GorillaKirsten V.K. Gilardi, Kristie L. Oxford,  David Gardner-Roberts,1 Jean-Felix Kinani,  Lucy Spelman,2 Peter A. Barry,  Michael R. Cranfield, and Linda J. LowenstineIn 2007, we detected human herpes simplex virus type 1,	which	caused	stomatitis,	in	a	juvenile	confiscated	eastern	lowland gorilla (Gorilla beringei graueri)	that	had	a	high	de-gree	of	direct	contact	with	human	caretakers.	Our	findings	confirm	that	pathogens	can	transfer	between	nonhuman	pri-mate hosts and humans.Most emerging infectious diseases of humans are of wildlife origin (1). The close genetic relatedness of humans and nonhuman primates (NHPs) makes each uniquely susceptible to pathogens of the other (2). Spill-over of NHP pathogens into humans can lead to human disease pandemics (e.g., HIV-1 from the chimpanzee strain of simian immunodeficiency virus [3]), and some NHP viruses are potentially acutely lethal to humans (e.g., herpes B virus of Asian macaques [4]). The opposite transmission event—human to NHP—is less frequently reported. Although of concern from a wildlife conserva-tion standpoint (5), human-to-NHP transmission events substantiate the fact that pathogen sharing is bidirectional (6). To provide further evidence for the potential for dis-ease emergence at the human–NHP interface, we report a human herpes simplex virus type 1 (HSV-1) infection in an eastern lowland gorilla (Grauer’s gorilla, Gorilla beringei graueri).The StudyA 2-year-old female Grauer’s gorilla, confiscated from poachers in Goma, Democratic Republic of the Congo, in November 2005, was group-housed in a facil-ity with 6 other juvenile Grauer’s gorillas and 1 juvenile mountain gorilla (G.b. beringei). All gorillas had daily direct contact with one another and with human caretak-ers who provided hands-on care. In March 2007, 5 of the Grauer’s gorillas exhibited bouts of lethargy and anorexia and oral lesions. Three of the affected gorillas, including the gorilla from Goma, were chemically immobilized for examination. All 3 gorillas exhibited multiple clear fluid-filled vesicles up to 2 cm in diameter that affected the mu-cosa of the lips and gingiva (Figure 1, panel A). Biopsy and swab samples were obtained from the lesions of the gorilla from Goma. The biopsy sample was fixed in 10% buffered formalin; the swab samples were stored frozen at –80°C, and these specimens were shipped to the United States for diagnostic testing.Histologic examination of the biopsy sample showed variably thickened, ulcerated, nonkeratinizing, strati-fied squamous oral mucosae. Margins of ulcers showed marked epidermal swelling (intracellular edema, balloon-ing degeneration) and large numbers of epithelial syncy-tial cells, often with marginated chromatin and smudgy intranuclear inclusion bodies filling the nucleus (Figure 1, panel B.). Inclusion bodies were present in individual cells in the stratum basalis and stratum spinosum adjacent to the ulcers. Using transmission electron microscopy, we observed virions consistent with herpesvirus in shape and size within the nucleus of syncytial cells and budding through the nuclear membrane (Figure 1, panel C.). His-topathology and transmission electron microscopy find-ings were essentially identical to findings from human HSV-1 infections (7).DNA was extracted from swab samples from this ani-mal (8) and amplified by PCR using pan-herpesvirus–spe-cific primers designed to the DNA polymerase gene (UL30) (9). A cloned and sequenced 731-nt amplicon (GenBank accession no. KJ396198) showed 99% identities with the DNA and protein sequences of HSV-1 UL30, and 94% and 95% identities (DNA and protein, respectively) to UL30 of HSV-2 (Table). Because sequences from gorilla-specif-ic herpesviruses have not been annotated, we performed a phylogenetic analysis of the amplicon from the gorilla swab sample using the UL30 genes from broadly repre-sentative isolates from the 3 herpesvirus subfamilies. The gorilla-derived DNA and protein (Figure 2) exhibited the strongest phylogenetic relationship with HSV-1. Align-ments of human and NHP alphaherpesvirus representa-tives show that the region amplified with the consensus herpesvirus primers is a highly conserved region of UL30 (Figure 2). The sequence alignments strongly suggested that the herpesvirus DNA in the oral lesion was HSV-1, although the gorilla-derived amplicon was an outlier in comparison with other HSV-1 isolates (data not shown). 	 Emerging	Infectious	Diseases	•	www.cdc.gov/eid	•	Vol.	20,	No.	11,	November	2014	 1883Author	 affiliations:	 University	 of	 California,	 Davis,	 California,	USA	 (K.V.K.	Gilardi,	K.L.	Oxford,	P.A.	Barry,	M.R.	Cranfield,	 L.J.	 Lowenstine);	and	Mountain	Gorilla	Veterinary	Project,	 Inc.,	Davis	(D.	Gardner-Roberts,	J.-F.	Kinani,	L.	Spelman,	M.R.	Cranfield)DOI:	http://dx.doi.org/10.3201/eid2011.1400751Current	 affiliation:	 ARMAC	 Veterinary	 Group,	 Biggar,	 South	Lanarkshire, Scotland, UK.2Current	 affiliation:	 Rhode	 Island	 School	 of	 Design,	 Providence,	Rhode Island, USA.The relative divergence of the amplicon was due primarily to nucleotide changes encoding a 3-aa motif not found in any other annotated sequence (Figure 2). Whether adapta-tion of the putative HSV-1 in the gorilla host represents a compensatory change to replication in the ectopic gorilla host is not known.To confirm our conclusion that the lesion was associ-ated with HSV-1 infection, we used HSV-1–specific prim-ers to amplify a region of UL30 (aa 134–246) outside of the amplicon defined by the pan-herpesvirus–specific prim-ers. Amplification was positive for the oral swab sample DNA with this second primer pair and for 3 human-isolated HSV-1 strains. Sequence analysis of the amplicon demon-strated 100% and 89% sequence identity with HSV-1 and HSV-2, respectively (data not shown).ConclusionOn the basis of the results of the histopathologic fea-tures and molecular screening of the virus, we concluded that the stomatitis in this juvenile female Grauer’s gorilla confiscated from poachers in Goma, Democratic Republic of the Congo, was caused by HSV-1. Alphaherpesvirus DISPATCHES1884	 Emerging	Infectious	Diseases	•	www.cdc.gov/eid	•	Vol.	20,No.	11,	November	2014Figure	1.	Vesicular	stomatitis	in	a	wild-caught	juvenile	Grauer’s	gorilla	(Gorilla beringei graueri).	Gross	lesions,	histopathologic	examination,	transmission	electron	microscopy,	and	molecular	screening	confirmed	human	herpesvirus	type	1	(HSV-1)	as	the	etiologic	agent.	A)	Human	HSV-1	lip	 lesions	in	a	wild-caught	 juvenile	Grauer’s	gorilla.	B)	Section	of	oral	mucosa	adjacent	to	a	vesiculo-ulcerative	 lesion	exhibits	epithelial	cell	necrosis,	cytoplasmic	swelling,	nuclear	chromatin	margination	 (sometimes	with	discrete	Cowdry	 type	A	 inclusions),	and	multinucleated	syncytia	typical	of	herpesviral	cytopathic	effects	(hematoxylin	and	eosin	stain).	Original	magnification	×200.	C)	Electron	micrograph of the same lesion demonstrates intranuclear, unenveloped virions ≈100 nm in diameter that are budding through the nuclear membrane to form enveloped virions ≈140	nm	 in	diameter;	morphologic	 features	of	both	are	compatible	with	a	herpesvirus.	Original	magnification	×60,000.	 Table. Comparison of gorilla amplicon to other herpesviruses* Gorilla	amplicon	identity HSV-1 HSV-2 MaHV-1 PaHV-2 HHV-3 HHV-4 HHV-5 HHV-6 HHV-8 EHV-1 DNA,	% 99.3 93.6 85.3 86.0 53.9 61.2 49.3 43.3 48.6 65.1 Protein, % 99.2 94.6 90.1 90.1 65.7 48.8 39.9 45.9 46.0 68.6 *Pairwise	alignment	of	the	gorilla	amplicon	DNA	sequence	(GenBank	accession	no.	KJ396198)	and	predicted	amino	acid	sequence	of the corresponding region	of	HSV-1	(HHV-1;	AFI98948),	HSV-2	(HHV-2;	AGI44412),	MaHV-1	(AAT67222);	PaHV-2 (YP_443877),	HHV-3	(varicella	zoster	virus;	ABF21820),	HHV-4	(Epstein-Barr	virus;	YP_401712),	HHV-5	(human	cytomegalovirus;	AAP37469),	HHV6	(BAF93477),	HHV-8	(Kaposi	sarcoma	virus;	ACY00400),	and	EHV-1	(ADI96155).	DNA–DNA	alignments	were	performed	by	using	the Wilbur-Lipman	Method	of	the	MegAlign	(DNAStar,	Madison,	WI,	USA)	sequence alignment program (Ktuple = 3;	gap	penalty = 3;	window = 20).	Protein–protein alignments were performed by using the Lipman-Pearson method (Ktuple = 2;	gap	penalty = 4;	gap	length	penalty = 12).	EHV,	equid	herpesvirus;	HSV,	herpes	simplex	virus;	HHV,	human	herpesvirus;	MaHV,	Macacine herpesvirus;	PaHV,	Papiine	herpesvirus.  HSV-1	in	Confiscated	Gorillaexposure in wild gorillas in this region has been reported (10), but to our knowledge, human HSV infection in wild-born captive gorillas has not been detected with molecu-lar techniques. Fatal HSV-1 infection in a captive-born, nursery-reared western lowland gorilla (G. gorilla goril-la) (11) has been reported. Serologic tests for HSV-1 on samples collected before the outbreak from other gorillas in the confiscated gorilla’s group showed that 2 clinically unaffected gorillas were seropositive for HSV-1, suggest-ing previous exposure to an alphaherpesvirus (12). How-ever, serologic tests for HSV-1 and -2 exposure in the 5 clinically affected gorillas during this outbreak, includ-ing the gorilla from Goma, were negative, which suggests these animals were experiencing their first HSV infec-tions, and antibodies were not yet detectable. Although the HSV infection status of the poachers who captured and initially held the gorilla from Goma was unknown, as was the human HSV infection status of the caretakers (from whom samples were not collected) who had daily contact with this gorilla and her captive cohort, we can safely assume most of these persons were representative of the general human population and therefore infected because HSV-1 seropositivity has been documented at a prevalence of >90% in the region (13–15), and HSV in-fections are lifelong.Our findings have major ramifications for the possible reintroduction of these confiscated gorillas into the wild (all remain in captivity) because such a management action might introduce a novel (human) pathogen to a naïve popu-lation. Screening wild Grauer’s gorillas and other gorilla populations for human HSV and other human pathogens will be critical to adequately assess the risk for reintroduc-ing captive gorillas to wild populations of these endangered great apes. From a global health standpoint, our finding of human HSV-1 in a gorilla proves once again that pathogens transfer among human and NHP hosts.AcknowledgmentsSamples were imported into the United States by permission of the Convention on International Trade in Endangered Species. We thank the Institut Congolais pour la Conservation de la Nature and the Rwanda Development Board (Antoine Mudakikwa) for permission to care for confiscated Grauer’s and mountain gorillas and the Dian Fossey Gorilla Fund for orphan gorilla management cooperation. Bob Nordhausen assisted with electron microscopy, and Christopher Polage provided human herpesvirus isolates.	 Emerging	Infectious	Diseases	•	www.cdc.gov/eid	•	Vol.	20,	No.	11,	November	2014	 1885Figure	2.	Phylogenetic	analysis	of	the	nucleotide	sequence	(A)	and	predicted	amino	acid	sequence	(B)	from	the	swab	sample	amplicon	from	the	gorilla	with	the	corresponding	regions	of	HSV-1	(HHV-1;	GenBank	accession	no.	AFI98948);	HSV-2	(HHV-2;	AGI44412);	MaHV-1	(AAT67222);	PaHV-2	(YP_443877);	HHV-3	(varicella	zoster	virus;	ABF21820);	HHV-4	(Epstein-Barr	virus;	YP_401712);	HHV-5	(human	cytomegalovirus;	AAP37469);	HHV-6	(BAF93477);	HHV-8	(Kaposi	sarcoma	virus;	ACY00400),	and	EHV-1	ADI96155).	Sequences	were	aligned	 by	 the	Clustal	W	method	 (http://www.clustal.org)	 by	 using	 the	MegAlign	 (DNAStar,	Madison,	WI,	 USA)	 sequence	 alignment	program	(multiple	alignment	parameters:	gap	penalty	=	15.00;	gap	length	penalty	=	6.66;	delay	divergent	seqs	(%)	=	30;	DNA	transition	weight	=	0.50.	Pairwise	alignment	parameters:	slow-accurate;	gap	penalty	=	15.00;	gap	length	=	6.66).	EHV,	equid	herpesvirus;	HSV,	herpes	simplex	virus;	HHV,	human	herpesvirus;	MaHV,	Macacine	herpesvirus;	PaHV,	Papiine	herpesvirus.This report was made possible in part by National Center for Research Resources and Office of Research Infrastructure Programs support to the California National Primate Research Center, and by the US Agency for International Development Emerging Pandemic Threats PREDICT project; contents do not necessarily reflect the views of USAID or the US Government.Dr Gilardi is co-director of the Karen C. Drayer Wildlife Health Center, School of Veterinary Medicine, University of Cal-ifornia, Davis. Her research interests include NHP medicine and conservation, zoonoses, marine wildlife and ecosystem health, and One Health.References  1. Jones KE, Patel NG, Levy MA, Storeygard A, Balk D, Gittleman JL, et al. Global trends in emerging infectious diseases. Nature. 2008;451:990–3. http://dx.doi.org/10.1038/nature06536  2. Sharp PM, Rayner JC, Hahn BH. Great apes and zoonoses. Science. 2013;340:284–6. http://dx.doi.org/10.1126/science.1236958  3. Gao F, Bailes E, Robertson DL, Chen Y, Rodenburg CM, Michael SF, et al. Origin of HIV-1 in the chimpanzee Pan troglodytes troglodytes. Nature. 1999;397:436–41. http://dx.doi.org/10.1038/17130  4. Huff JL, Barry PA. B-virus (Cercopithecine herpesvirus 1) infec-tion in humans and macaques: potential for zoonotic disease. Emerg Infect Dis. 2003;9:246–50. http://dx.doi.org/10.3201/eid0902.020272  5. Palacios G, Lowenstine LJ, Cranfield MR, Gilardi KVK, Spelman L, Lukasik-Braum M, et al. Human metapneumovirus infection in wild mountain gorillas in Rwanda. Emerg Infect Dis. 2011;17:711–3. http://dx.doi.org/10.3201/eid1704.100883  6. Chen EC, Yagi S, Kelly KR, Mendoza SP, Maninger N, Rosenthal A, et al. Cross-species transmission of a novel adenovirus associated with a fulminant pneumonia outbreak in a New World monkey colony. PLoS Pathog. 2011;7:e1002155. Erratum in: PLoS Pathog. 2011;7. http://dx.doi.org/10.1371/journal.ppat.1002155  7. Chisholm C, Lopez L. Cutaneous infections caused by Herpes-viridae: a review. Arch Pathol Lab Med. 2011;135:1357–62. http://dx.doi.org/10.5858/arpa.2010-0156-RS  8. Oxford KL, Eberhardt MK, Yang KW, Strelow L, Kelly S, Zhou SS, et al. Protein coding content of the ULb′ region of wild-type rhesus cytomegalovirus. Virology. 2008;373:181–8. http://dx.doi.org/10.1016/j.virol.2007.10.040  9. Ehlers B, Borchers K, Grund C, Frolich K, Ludwig H, Buhk HJ. Detection of new DNA polymerase genes of known and potentially novel herpesviruses by PCR with degenerate and deoxyinosine-substituted primers. Virus Genes. 1999;18:211–20. http://dx.doi.org/10.1023/A:100806411805710. Eberle R. Evidence for an alpha-herpesvirus indigenous to mountain gorillas. J Med Primatol. 1992;21:246–51.11. Heldstab A, Ruedi D, Sonnabend W, Deinhardt F. Spontaneous gen-eralized Herpesvirus hominis infection of a lowland gorilla (Gorilla gorilla gorilla). J Med Primatol. 1981;10:129–35.12. Whittier C, Nutter FB, Cranfield MR. Seroprevalence of infec-tious agents in free-living mountain gorillas (Gorilla beringei spp). Proceedings of the Wildlife Disease Association International Con-ference 2005, Cairns, Australia. Abstract 174, p. 291.13. Wagner HU, Van Dyck E, Roggen E, Nunn AJ, Kamali A, Schmid DS, et al. Seroprevalence and incidence of sexually-transmitted diseases in a rural Ugandan population. Int J STD AIDS. 1994;5:332–7.14. Kasubi MJ, Nilson A, Marsden HS, Bergstrom T, Langeland N, Haarr L. Prevalence of antibodies against herpes simplex virus types 1 and 2 in children and young people in an urban region in Tan-zania. J Clin Microbiol. 2006;44:2801–7. http://dx.doi.org/10.1128/JCM.00180-0615. Mountain Gorilla Veterinary Project 2002 Employee Health Group. Risk of disease transmission between conservation personnel and the mountain gorillas: results from an employee health program in Rwanda. EcoHealth. 2004;1:351–61. http://dx.doi.org/10.1007/s10393-004-0116-4Address for correspondence: Kirsten V.K. Gilardi, Wildlife Health Center, University of California, 1089 Veterinary Medicine Dr, Davis, CA 95614, USA; email: kvgilardi@ucdavis.eduDISPATCHES1886	 Emerging	Infectious	Diseases	•	www.cdc.gov/eid	•	Vol.	20,No.	11,	November	2014Online subscription: wwwnc.cdc.gov/eid/subscribe.htm

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Human Herpes Simplex Virus Type 1 in Confiscated Gorilla - Volume 20, Number 11—November 2014 - Emerging Infectious Diseases journal - CDC

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Human herpes simplex virus type 1 in confiscated gorilla.

Human herpes simplex virus type 1 in confiscated gorilla.

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