Orf Virus Infection in Alaskan Mountain Goats, Dall's Sheep, Muskoxen
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Tryland et al. Acta Vet Scand (2018) 60:12 https://doi.org/10.1186/s13028-018-0366-8 Acta Veterinaria Scandinavica RESEARCH Open Access Orf virus infection in Alaskan mountain goats, Dall’s sheep, muskoxen, caribou and Sitka black‑tailed deer Morten Tryland1* , Kimberlee Beth Beckmen2, Kathleen Ann Burek‑Huntington3, Eva Marie Breines1 and Joern Klein4* Abstract Background: The zoonotic Orf virus (ORFV; genus Parapoxvirus, Poxviridae family) occurs worldwide and is transmit‑ ted between sheep and goats, wildlife and man. Archived tissue samples from 16 Alaskan wildlife cases, representing mountain goat (Oreamnos americanus, n 8), Dall’s sheep (Ovis dalli dalli, n 3), muskox (Ovibos moschatus, n 3), Sitka black-tailed deer (Odocoileus hemionus= sitkensis, n 1) and caribou (Rangifer= tarandus granti, n 1), were analyzed.= = = Results: Clinical signs and pathology were most severe in mountain goats, afecting most mucocutaneous regions, including palpebrae, nares, lips, anus, prepuce or vulva, as well as coronary bands. The proliferative masses were solid and nodular, covered by dark friable crusts. For Dall’s sheep lambs and juveniles, the gross lesions were similar to those of mountain goats, but not as extensive. The muskoxen displayed ulcerative lesions on the legs. The caribou had two ulcerative lesions on the upper lip, as well as lesions on the distal part of the legs, around the main and dew claws. A large hairless spherical mass, with the characteristics of a fbroma, was sampled from a Sitka black-tailed deer, which did not show proliferative lesions typical of an ORFV infection. Polymerase chain reaction analyses for B2L, GIF, vIL-10 and ATI demonstrated ORFV specifc DNA in all cases. Sequences from Dall’s sheep formed a separate cluster, comparable to ORFV from domestic sheep. Sequences from the other species were diferent from the Dall’s sheep sequences, but almost identical to each other. Conclusions: This is the frst major investigation of parapoxvirus infections in large Alaskan game species, and the frst report of parapoxvirus infection in caribou and Sitka black-tailed deer. This study shows that most of the wild ruminant species in Alaska and from most parts of Alaska, can carry and be afected by ORFV. These fndings call for attention to transmission of ORFV from wildlife to livestock and to hunters, subsistence harvesters, and wildlife biologists. Keywords: Alaska, Caribou, Contagious ecthyma, Dall’s sheep, Deer, Mountain goat, Muskox, Parapoxvirus, Virology, Wildlife, Zoonosis Background and Parapoxvirus of red deer in New Zealand [1]. ORFV Orf virus (ORFV) is the type species of the genus Para- is distributed worldwide with domestic sheep and goats, poxvirus of the Poxviridae family, which also includes causing contagious ecthyma (CE) (syn. contagious pustu- Bovine papular stomatitis virus, Pseudocowpox virus lar dermatitis, scabby mouth). ORFV produce lesions in the skin and at muco-cutaneous interface at the muzzle, lips, eyelids, anus, prepuce or vulva, and teats as well as *Correspondence: [email protected]; [email protected] in the oral mucosa, such as the tongue and the gingiva, 1 Department of Arctic and Marine Biology, Arctic Infection Biology, UiT- especially associated with erupting teeth [2]. CE typically Arctic University of Norway, Framstredet 39, 9019 Tromsø, Norway 4 Department of Nursing and Health Sciences‑Vestfold, Faculty starts with a macule at the port of entry, progressing to of Health and Social Sciences, University College of Southeast Norway, a papule and pustule. Since ORFV express the proteins 3603 Kongsberg, Norway homologous to endothelial growth factor and vascular Full list of author information is available at the end of the article © The Author(s) 2018. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/ publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Tryland et al. Acta Vet Scand (2018) 60:12 Page 2 of 11 permeability factor, both being angiogenesis regulators Te aims of this study were to verify the preliminary CE [3], such lesions becomes highly vascularized and pro- diagnoses in 16 wildlife cases by identifying the causative liferative (“caulifower-like”) and are easily exposed to virus. We also wanted to characterize this zoonotic dis- trauma and bleeding, followed by secondary bacterial ease in wild ruminant game species in Alaska, and con- infections. Chronic CE lesions are thus often presented duct a phylogenetic comparison of the circulating viruses as masses covered by thick crusts (skin) or as prolifera- with known reference viruses of the Parapoxvirus genus. tive masses (mucosa). Histological changes are present in the epithelium as ballooning degeneration, reticular Methods degeneration, marked epidermal proliferation and epi- Clinical cases and sampling dermal micro-abscesses [4]. Fifteen animals (Table 1) that during the period 2002– CE, often referred to as “contagious ecthyma-like” 2012 were hunted, reported dead or euthanized for when lacking a virological confrmation, has been animal welfare reasons were subjected to our investiga- reported in many free ranging animal species [5], such tions. Tey had lesions in the mucocutaneous interface as semi-domesticated reindeer (Rangifer tarandus taran- and/or on coronary bands consistent with or suspicious dus) [6–8], captive and wild muskoxen (Ovibos moscha- of CE. One additional animal (Sitka black-tailed deer) tus) [9–11], and red deer (Cervus elaphus) [12, 13]. had lesions consistent with a fbroma, although CE was In Alaska, CE has been described in the domestic sheep mentioned as a diferential diagnosis in the necropsy and goats, as well as in wild Dall’s sheep (Ovis dalli dalli), report. Te geographical location of the clinical cases is mountain goat (Oreamnos americanus) and muskox [9, depicted in Fig. 1. Te animals were subjected to feld 14]. Severe outbreaks have also been reported in Alaskan dressing, and whole carcasses or body parts were sent to captive muskox [15], and it has been shown experimen- Alaska Department of Fish and Game, Fairbanks, Alaska, tally that also moose (Alces alces) and caribou (Rangifer for diagnostic investigations. Tissue samples from gross tarandus) calves are susceptible to infection with ORFV lesions were obtained during necropsies, using aseptic isolated from sheep [9]. techniques and stored in Whirl Pac (Nasco, Medesto CA, Since ORFV is zoonotic, these infections represent an USA) or cryogenic vials at − 20 °C or colder. A subset of occupational hazard for people handling afected ani- these frozen tissue samples (20 mg each) was obtained mals, such as veterinarians, sheep herders, hunters and from three diferent areas from each case, from areas game managers [16–18]. In the scientifc literature, one showing gross pathological changes consistent with CE, human case has been reported from Alaska, with ORFV using aseptic techniques to avoid contamination between transmitted from a mountain goat [19]. ORFV is prob- samples. Tese subsamples were subjected to further ably one of the most common poxvirus infection in virological investigations. Additional tissue sections were humans, causing lesions mainly on fngers and hands. preserved in 10% neutral bufered formalin. In most patients, a single papule appears which may develop into a large vesicle or bulla, sometimes being Histopathology hemorrhagic, with a black appearance. Te condition can Formalin fxed tissues were processed routinely, and par- be associated with by local lymphadenopathy and mild afn-embedded sections were cut to 5 μm, and stained fever and is usually mildly painful, but ORFV infections with hematoxylin and eosin at Histology Consulting Ser- may also become generalized and severe, especially in vices Inc. (Everson, WA, USA). immunocompromised persons [20]. Humans are infected through direct contact with infected animals, including Polymerase chain reaction (PCR) and sequencing wildlife [16, 19]. ORFV infection is well known among DNA was extracted (Alaska Dept. of Fish and Game, farmers and the disease is listed as one that people of the USA) from the tissue samples using QIAamp ® DNA Mini Arctic should be aware of through contact with infected Kit (QIAGEN, Hilden, Germany), with an average output game species [21]. of 94 µg/mL (29–253 µg/mL). Four diferent PCR proto- A large number of game is harvested in Alaska each cols were performed (UiT-Arctic University of Norway) year, by indigenous people as part of their tradition as specifed in Table 2. Extracted DNA from tissue from and culture, and by local and non-resident hunters. In a sheep with CE and verifed ORFV infection was used as 2015, 15,554 Sitka black-tailed deer, 4332 caribou, 256 positive control. Deionized and fltered water was used as muskoxen, 528 mountain goats, and 11 Dall’s sheep were a negative control. reported harvested in Alaska [22], indicating that people, Te PCRs were run in a DNA Termal Cycler PE9700 to a substantial degree, are in close contact with these (PerkinElmer Norge AS, Kristiansand, Norway). Te large game species. PPP 1 and PPP 4 primers targeting the