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Morten Tryland - One of the best experts on this subject based on the ideXlab platform.
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Orf virus infection in Alaskan mountain goats, Dall’s sheep, muskoxen, caribou and Sitka black-tailed deer
BMC, 2018Co-Authors: Morten Tryland, Kimberlee Beth Beckmen, Kathleen Ann Burek-huntington, Eva Marie Breines, Joern KleinAbstract:Abstract Background The zoonotic Orf virus (ORFV; genus Parapoxvirus, Poxviridae family) occurs worldwide and is transmitted 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, affecting 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 fibroma, 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 specific 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 different from the Dall’s sheep sequences, but almost identical to each other. Conclusions This is the first major investigation of Parapoxvirus infections in large Alaskan game species, and the first 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 affected by ORFV. These findings call for attention to transmission of ORFV from wildlife to livestock and to hunters, subsistence harvesters, and wildlife biologists
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A severe outbreak of contagious ecthyma (orf) in a free-ranging musk ox (Ovibos moschatus) population in Norway.
Veterinary microbiology, 2007Co-Authors: Turid Vikøren, Tord Bretten, Atle Lillehaug, Johan Akerstedt, Magne Haugum, Morten TrylandAbstract:During July-October 2004, 19 (18 calves, 1 yearling) free-ranging musk oxen (Ovibos moschatus) at Dovre, Norway, were observed with contagious echtyma-like lesions, and 16 of them were euthanized. Six musk oxen were subjected to necropsy, histopathological and microbiological examinations. All euthanized animals had lesions consistent with contagious ecthyma presenting as wart-like, scabby lesions on the muzzle, lips, oral mucosa and limbs to a variable extent. The histopathological examination showed pustular dermatitis characterized by epidermal proliferation, reticular degeneration, degenerating keratinocytes with intracytoplasmic eosinophilic inclusion bodies, vesicopustules, microabscesses and multifocal ulcerations in the epidermis which was covered by a serocellular crust. Pathology and bacteriology showed evidence of secondary infections in the skin and draining lymph nodes. Electron microscopy (negative staining) of lesions from four animals detected Parapoxvirus with the typical arrangement of the outer protein filaments. Parapoxvirus DNA was detected in tissue samples from two examined animals by polymerase chain reaction (PCR) with primers from the B2L-gene. A DNA sequence of 326 nucleotides from the amplicon was compared with similar DNA sequences from Parapoxvirus isolated from sheep, reindeer, musk ox and cattle. The outbreak was caused by a virus similar to other circulating orf virus variants in Norway. Antibodies against Parapoxvirus were detected with a virus neutralization test in 3 of 35 musk oxen (8.6%) sampled at Dovre between 2004 and 2006. This is the first report of a severe outbreak of contagious ecthyma in free-ranging musk oxen.
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Characterisation of Parapoxviruses isolated from Norwegian semi-domesticated reindeer (Rangifer tarandus tarandus)
Virology Journal, 2005Co-Authors: Joern Klein, Morten TrylandAbstract:Background Two outbreaks of the disease contagious ecthyma were reported in 1999 and 2000 in Norwegian semi-domesticated reindeer ( Rangifer tarandus tarandus ). Contagious ecthyma is an epidermal disease of sheep and goats worldwide, which is caused by the zoonotic Parapoxvirus orf virus. Characterisation of clinical samples from the two outbreaks in semi-domesticated reindeer in Norway by electron microscopy and PCR (B2L) revealed typical Parapoxvirus particles and partial gene sequences corresponding to Parapoxvirus, respectively. If contagious ecthyma in reindeer is caused by orf virus, the virus may be transferred from sheep and goats, via people, equipment and common use of pastures and corrals, to reindeer. Another possibility is that contagious ecthyma in reindeer is caused by a hitherto unclassified member of the Parapoxvirus genus that circulates among reindeer herds and remains endemic in Norway. Results Genomic comparisons of one standard orf strain (orf NZ2) and the reindeer isolates, employing restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) analysis, demonstrated high similarity between the reindeer viruses and known orf virus strains. Partial DNA sequences of two different viral genes were determined for the different isolates and compared with corresponding Parapoxvirus genebank sequences. The comparison/alignment and construction of phylogenetic trees also point to an affiliation of the reindeer viruses to the species orf virus. Conclusion The results of this work imply that the Parapoxvirus causing contagious ecthyma in Norwegian semi-domesticated reindeer belongs to the species orf virus and that the orf virus crosses the host species barrier from sheep and goat to semi-domesticated reindeer.
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characterisation of Parapoxviruses isolated from norwegian semi domesticated reindeer rangifer tarandus tarandus
Virology Journal, 2005Co-Authors: Joern Klein, Morten TrylandAbstract:Background Two outbreaks of the disease contagious ecthyma were reported in 1999 and 2000 in Norwegian semi-domesticated reindeer (Rangifer tarandus tarandus). Contagious ecthyma is an epidermal disease of sheep and goats worldwide, which is caused by the zoonotic Parapoxvirus orf virus. Characterisation of clinical samples from the two outbreaks in semi-domesticated reindeer in Norway by electron microscopy and PCR (B2L) revealed typical Parapoxvirus particles and partial gene sequences corresponding to Parapoxvirus, respectively. If contagious ecthyma in reindeer is caused by orf virus, the virus may be transferred from sheep and goats, via people, equipment and common use of pastures and corrals, to reindeer. Another possibility is that contagious ecthyma in reindeer is caused by a hitherto unclassified member of the Parapoxvirus genus that circulates among reindeer herds and remains endemic in Norway.
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Isolation and partial characterization of a Parapoxvirus isolated from a skin lesion of a Weddell seal
Virus Research, 2005Co-Authors: Morten Tryland, Joern Klein, Erling S. Nordøy, Arnoldus Schytte BlixAbstract:Abstract A solitary skin lesion was found on the neck of a Weddell seal ( Leptonychotes weddellii ), chemically immobilized in Queen Maud Land (70°09′S, 05°22′E) Antarctica 2001. The lesion was elevated and 3 cm in diameter, consisting of partly fresh and partly necrotic tissue, and proliferative papilloma-like structures were seen. Electron microscopy on a biopsy from the lesion revealed typical Parapoxvirus particles. Polymerase chain reaction (PCR; B2L gene) generated amplicons of approximately 594 base pairs, comparable to Orf-virus, the prototype Parapoxvirus. A comparison of these B2L PCR amplicon DNA sequences with corresponding sequences from other Parapoxviruses, showed that the Weddell seal virus resembled isolates from grey seal ( Halichoerus grypus ) and harbour seal ( Phoca vitulina ) more than Parapoxvirus from red deer ( Cervus elaphus ), sheep, cattle and Japanese serows ( Capricornis crispus ). It is thus concluded that the Weddell seal Parapoxvirus belong to the tentative seal Parapoxvirus species. Since parapox and orthopoxviruses may cause similar clinical diseases, we suggest that the term sealpox should be restricted to the clinical disease, whereas seal Parapoxvirus should be used when caused by a Parapoxvirus, rather than the general term “sealpox virus”. This is the first verified case of Parapoxvirus infection in a Weddell seal, and also the first report of any such infections in the Antarctic.
Joern Klein - One of the best experts on this subject based on the ideXlab platform.
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Orf virus infection in Alaskan mountain goats, Dall’s sheep, muskoxen, caribou and Sitka black-tailed deer
BMC, 2018Co-Authors: Morten Tryland, Kimberlee Beth Beckmen, Kathleen Ann Burek-huntington, Eva Marie Breines, Joern KleinAbstract:Abstract Background The zoonotic Orf virus (ORFV; genus Parapoxvirus, Poxviridae family) occurs worldwide and is transmitted 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, affecting 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 fibroma, 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 specific 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 different from the Dall’s sheep sequences, but almost identical to each other. Conclusions This is the first major investigation of Parapoxvirus infections in large Alaskan game species, and the first 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 affected by ORFV. These findings call for attention to transmission of ORFV from wildlife to livestock and to hunters, subsistence harvesters, and wildlife biologists
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characterisation of Parapoxviruses isolated from norwegian semi domesticated reindeer rangifer tarandus tarandus
Virology Journal, 2005Co-Authors: Joern Klein, Morten TrylandAbstract:Background Two outbreaks of the disease contagious ecthyma were reported in 1999 and 2000 in Norwegian semi-domesticated reindeer (Rangifer tarandus tarandus). Contagious ecthyma is an epidermal disease of sheep and goats worldwide, which is caused by the zoonotic Parapoxvirus orf virus. Characterisation of clinical samples from the two outbreaks in semi-domesticated reindeer in Norway by electron microscopy and PCR (B2L) revealed typical Parapoxvirus particles and partial gene sequences corresponding to Parapoxvirus, respectively. If contagious ecthyma in reindeer is caused by orf virus, the virus may be transferred from sheep and goats, via people, equipment and common use of pastures and corrals, to reindeer. Another possibility is that contagious ecthyma in reindeer is caused by a hitherto unclassified member of the Parapoxvirus genus that circulates among reindeer herds and remains endemic in Norway.
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Characterisation of Parapoxviruses isolated from Norwegian semi-domesticated reindeer (Rangifer tarandus tarandus)
Virology Journal, 2005Co-Authors: Joern Klein, Morten TrylandAbstract:Background Two outbreaks of the disease contagious ecthyma were reported in 1999 and 2000 in Norwegian semi-domesticated reindeer ( Rangifer tarandus tarandus ). Contagious ecthyma is an epidermal disease of sheep and goats worldwide, which is caused by the zoonotic Parapoxvirus orf virus. Characterisation of clinical samples from the two outbreaks in semi-domesticated reindeer in Norway by electron microscopy and PCR (B2L) revealed typical Parapoxvirus particles and partial gene sequences corresponding to Parapoxvirus, respectively. If contagious ecthyma in reindeer is caused by orf virus, the virus may be transferred from sheep and goats, via people, equipment and common use of pastures and corrals, to reindeer. Another possibility is that contagious ecthyma in reindeer is caused by a hitherto unclassified member of the Parapoxvirus genus that circulates among reindeer herds and remains endemic in Norway. Results Genomic comparisons of one standard orf strain (orf NZ2) and the reindeer isolates, employing restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) analysis, demonstrated high similarity between the reindeer viruses and known orf virus strains. Partial DNA sequences of two different viral genes were determined for the different isolates and compared with corresponding Parapoxvirus genebank sequences. The comparison/alignment and construction of phylogenetic trees also point to an affiliation of the reindeer viruses to the species orf virus. Conclusion The results of this work imply that the Parapoxvirus causing contagious ecthyma in Norwegian semi-domesticated reindeer belongs to the species orf virus and that the orf virus crosses the host species barrier from sheep and goat to semi-domesticated reindeer.
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Isolation and partial characterization of a Parapoxvirus isolated from a skin lesion of a Weddell seal
Virus Research, 2005Co-Authors: Morten Tryland, Joern Klein, Erling S. Nordøy, Arnoldus Schytte BlixAbstract:Abstract A solitary skin lesion was found on the neck of a Weddell seal ( Leptonychotes weddellii ), chemically immobilized in Queen Maud Land (70°09′S, 05°22′E) Antarctica 2001. The lesion was elevated and 3 cm in diameter, consisting of partly fresh and partly necrotic tissue, and proliferative papilloma-like structures were seen. Electron microscopy on a biopsy from the lesion revealed typical Parapoxvirus particles. Polymerase chain reaction (PCR; B2L gene) generated amplicons of approximately 594 base pairs, comparable to Orf-virus, the prototype Parapoxvirus. A comparison of these B2L PCR amplicon DNA sequences with corresponding sequences from other Parapoxviruses, showed that the Weddell seal virus resembled isolates from grey seal ( Halichoerus grypus ) and harbour seal ( Phoca vitulina ) more than Parapoxvirus from red deer ( Cervus elaphus ), sheep, cattle and Japanese serows ( Capricornis crispus ). It is thus concluded that the Weddell seal Parapoxvirus belong to the tentative seal Parapoxvirus species. Since parapox and orthopoxviruses may cause similar clinical diseases, we suggest that the term sealpox should be restricted to the clinical disease, whereas seal Parapoxvirus should be used when caused by a Parapoxvirus, rather than the general term “sealpox virus”. This is the first verified case of Parapoxvirus infection in a Weddell seal, and also the first report of any such infections in the Antarctic.
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Isolation and partial characterization of a Parapoxvirus isolated from a skin lesion of a Weddell seal.
Virus research, 2005Co-Authors: Morten Tryland, Joern Klein, Erling S. Nordøy, Arnoldus Schytte BlixAbstract:A solitary skin lesion was found on the neck of a Weddell seal (Leptonychotes weddellii), chemically immobilized in Queen Maud Land (70 degrees 09'S, 05 degrees 22'E) Antarctica 2001. The lesion was elevated and 3cm in diameter, consisting of partly fresh and partly necrotic tissue, and proliferative papilloma-like structures were seen. Electron microscopy on a biopsy from the lesion revealed typical Parapoxvirus particles. Polymerase chain reaction (PCR; B2L gene) generated amplicons of approximately 594 base pairs, comparable to Orf-virus, the prototype Parapoxvirus. A comparison of these B2L PCR amplicon DNA sequences with corresponding sequences from other Parapoxviruses, showed that the Weddell seal virus resembled isolates from grey seal (Halichoerus grypus) and harbour seal (Phoca vitulina) more than Parapoxvirus from red deer (Cervus elaphus), sheep, cattle and Japanese serows (Capricornis crispus). It is thus concluded that the Weddell seal Parapoxvirus belong to the tentative seal Parapoxvirus species. Since parapox and orthopoxviruses may cause similar clinical diseases, we suggest that the term sealpox should be restricted to the clinical disease, whereas seal Parapoxvirus should be used when caused by a Parapoxvirus, rather than the general term "sealpox virus". This is the first verified case of Parapoxvirus infection in a Weddell seal, and also the first report of any such infections in the Antarctic.
Antonio Lavazza - One of the best experts on this subject based on the ideXlab platform.
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Parapoxvirus infections of red deer, Italy
Emerging Infectious Diseases, 2011Co-Authors: Alessandra Scagliarini, Franco Vaccari, Filippo Turrini, Alessandro Bianchi, Paolo Cordioli, Antonio LavazzaAbstract:The genus Parapoxvirus (family Poxviridae, subfamily Chordopoxvirinae) comprises several members: orf virus (OV), bovine papular stomatitis virus (BPSV), pseudocowpox virus (PCPV), and parapox of red deer in New Zealand virus (PVNZ). PVNZ is responsible for a contagious pustular dermatitis in farmed red deer, with outbreaks reported only in New Zealand (1). Cases of Parapoxvirus (PPV) pustular stomatitis were reported in wild ruminants in Stelvio Park in the Italian Alps during 2008. The affected animals had erosions and ulcers in the mouth, which led to death by starvation, particularly during the winter. Similar cases have also been described during 1992 in Finland and Norway in reindeer (Rangifer tarandus). Recently, the causative viruses of the clinical forms in reindeer were shown to be closely related to OV virus and PCPV, excluding the circulation of PVNZ in these countries (2,3). To characterize the PPV agents causing severe disease in wild ruminants of Stelvio Park, we sequenced and compared the DNA of several isolates. Results showed that the viruses isolated from chamois (Rupicapra rupicapra) and ibex (Capra ibex) were closely related to OV, whereas the isolates from red deer (Cervus elaphus) grouped with PVNZ. Our fi ndings provide new information about the diffusion of PPVs in wild ruminants and evidence that PVNZ is circulating outside New Zealand.
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Parapoxvirus infections of red deer italy
Emerging Infectious Diseases, 2011Co-Authors: Alessandra Scagliarini, Filippo Turrini, Alessandro Bianchi, Paolo Cordioli, Francesca Vaccari, Antonio LavazzaAbstract:To characterize Parapoxviruses causing severe disease in wild ruminants in Stelvio Park, Italy, we sequenced and compared the DNA of several isolates. Results demonstrated that the red deer isolates are closely related to the parapox of red deer in New Zealand virus.
Arnoldus Schytte Blix - One of the best experts on this subject based on the ideXlab platform.
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Isolation and partial characterization of a Parapoxvirus isolated from a skin lesion of a Weddell seal
Virus Research, 2005Co-Authors: Morten Tryland, Joern Klein, Erling S. Nordøy, Arnoldus Schytte BlixAbstract:Abstract A solitary skin lesion was found on the neck of a Weddell seal ( Leptonychotes weddellii ), chemically immobilized in Queen Maud Land (70°09′S, 05°22′E) Antarctica 2001. The lesion was elevated and 3 cm in diameter, consisting of partly fresh and partly necrotic tissue, and proliferative papilloma-like structures were seen. Electron microscopy on a biopsy from the lesion revealed typical Parapoxvirus particles. Polymerase chain reaction (PCR; B2L gene) generated amplicons of approximately 594 base pairs, comparable to Orf-virus, the prototype Parapoxvirus. A comparison of these B2L PCR amplicon DNA sequences with corresponding sequences from other Parapoxviruses, showed that the Weddell seal virus resembled isolates from grey seal ( Halichoerus grypus ) and harbour seal ( Phoca vitulina ) more than Parapoxvirus from red deer ( Cervus elaphus ), sheep, cattle and Japanese serows ( Capricornis crispus ). It is thus concluded that the Weddell seal Parapoxvirus belong to the tentative seal Parapoxvirus species. Since parapox and orthopoxviruses may cause similar clinical diseases, we suggest that the term sealpox should be restricted to the clinical disease, whereas seal Parapoxvirus should be used when caused by a Parapoxvirus, rather than the general term “sealpox virus”. This is the first verified case of Parapoxvirus infection in a Weddell seal, and also the first report of any such infections in the Antarctic.
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Isolation and partial characterization of a Parapoxvirus isolated from a skin lesion of a Weddell seal.
Virus research, 2005Co-Authors: Morten Tryland, Joern Klein, Erling S. Nordøy, Arnoldus Schytte BlixAbstract:A solitary skin lesion was found on the neck of a Weddell seal (Leptonychotes weddellii), chemically immobilized in Queen Maud Land (70 degrees 09'S, 05 degrees 22'E) Antarctica 2001. The lesion was elevated and 3cm in diameter, consisting of partly fresh and partly necrotic tissue, and proliferative papilloma-like structures were seen. Electron microscopy on a biopsy from the lesion revealed typical Parapoxvirus particles. Polymerase chain reaction (PCR; B2L gene) generated amplicons of approximately 594 base pairs, comparable to Orf-virus, the prototype Parapoxvirus. A comparison of these B2L PCR amplicon DNA sequences with corresponding sequences from other Parapoxviruses, showed that the Weddell seal virus resembled isolates from grey seal (Halichoerus grypus) and harbour seal (Phoca vitulina) more than Parapoxvirus from red deer (Cervus elaphus), sheep, cattle and Japanese serows (Capricornis crispus). It is thus concluded that the Weddell seal Parapoxvirus belong to the tentative seal Parapoxvirus species. Since parapox and orthopoxviruses may cause similar clinical diseases, we suggest that the term sealpox should be restricted to the clinical disease, whereas seal Parapoxvirus should be used when caused by a Parapoxvirus, rather than the general term "sealpox virus". This is the first verified case of Parapoxvirus infection in a Weddell seal, and also the first report of any such infections in the Antarctic.
Hiroshi Sentsui - One of the best experts on this subject based on the ideXlab platform.
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Persistent Parapoxvirus infection in cattle.
Microbiology and immunology, 2002Co-Authors: Yuko Iketani, Yasuo Inoshima, Shinya Shimizu, Kenji Murakami, Akihiro Asano, Hiroshi SentsuiAbstract:The possibility of persistent Parapoxvirus (PPV) infection was investigated by serologically and genetically using cattle infected with the virus experimentally and naturally. Three cattle were inoculated with the virus subcutaneously at several spots in the lips and abdominal regions. Small papules developed in the inoculated regions, and antibodies to the virus developed and continued persistently. One animal, from which one PPV had been previously isolated, was also subjected to serological and viral detection tests as a naturally infected case. Two of these four cattle were injected with dexamethasone (DM), and one was injected with interferon-γ (IFN-γ). The viral genome was rarely detected from the peripheral blood leukocytes in the ordinary condition, but frequently when the animals were injected with IFN-γ. The viral genome was also detected from the lymph nodes as these PPV infected animals were euthanized. These results indicated that cattle were infected with PPV subclinically and persistently, and the virus was activated in stressed or immunosuppressed animals. The virus would be harbored in the lymphotic tissues of the animals when they show no clinical symptoms.
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Serological survey of Parapoxvirus infection in wild ruminants in Japan in 1996-9.
Epidemiology and infection, 2001Co-Authors: Yasuo Inoshima, Shinya Shimizu, Y. Yamamoto, T. Takahashi, M. Shino, A. Katsumi, Hiroshi SentsuiAbstract:The prevalence of Parapoxvirus infection was examined in free-ranging wild ruminants in Japan, Japanese serow (Capricornis crispus) and Japanese deer (Cervus nippon centralis), in 1996-9. We collected a total of 151 serum samples from 101 Japanese serows and 50 Japanese deer and tested for antibodies against Parapoxvirus by an enzyme-linked immunosorbent assay and an agar gel immunodiffusion test. Overall seroprevalences among Japanese serows were 5/25 (20.0%) in 1996, 4/14 (28.6%) in 1997, 5/32 (15.6%) in 1998 and 2/30 (6.7%) in 1999, respectively. The seroprevalence increased with age but was not affected by sex. No antibodies were detected from any of 50 serum samples taken from Japanese deer. Our results in this study suggest that Parapoxvirus infection is widespread among the population of Japanese serows, however, Japanese deer appear to be still free of the disease.
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Detection and diagnosis of Parapoxvirus by the polymerase chain reaction.
Journal of virological methods, 2000Co-Authors: Yasuo Inoshima, Akira Morooka, Hiroshi SentsuiAbstract:The genus Parapoxvirus includes four members, bovine papular stomatitis virus (BPSV), pseudocowpox virus (PCPV), orf virus (ORFV) and Parapoxvirus of red deer in New Zealand (PVNZ). A set of primers for polymerase chain reaction (PCR) was designed to detect viral DNA from cells infected with each of the four Parapoxviruses. The set of primers resulted in the amplification of appropriately sized products from cells infected with BPSV, PCPV, ORFV and PVNZ, respectively. The PCR method was applied for the detection of seven field isolates of Parapoxvirus from cattle, sheep and free-ranging wild Japanese serows. The expected size of DNA was amplified from cells infected with each of the seven isolates. No specific PCR products were detected from vaccinia virus-, fowlpox virus- and mock-infected cells. Moreover, by a semi-nested PCR with an inner primer and Southern blot analysis, viral DNA was detected from lesions of clinically affected cattle, sheep and Japanese serows. These results suggested that the PCR method used in this study was specific for the detection of Parapoxviruses and thus useful for diagnosis of Parapoxvirus infections, especially in discrimination from diseases with similar clinical symptoms.
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Simple preparation of Parapoxvirus genome DNA for endonuclease analysis.
Microbiology and immunology, 2000Co-Authors: Yasuo Inoshima, Kenji Murakami, Akira Morooka, Hiroshi SentsuiAbstract:We compared five methods for improved extraction of very-large Parapoxvirus DNA from infected cells: (i) alkaline-lysis procedure followed by phenol extraction; (ii) modified Hirt procedure, which was a neutral lysis procedure followed by phenol extraction; (iii) Hirt procedure; (iv) method used for extraction of vaccinia virus DNA; and (v) standard procedure using virus purification with an ultracentrifuge and protease-sodium dodecyl sulfate-phenol treatment. The alkaline-lysis procedure was more rapid, inexpensive and simpler than the other methods. Moreover, with this method it is not necessary to prepare any special facilities, reagents and kits. Although the extracted DNA was still crude, we could reproducibly prepare viral DNA from 2 × 106 infected cells in less than 2 hr and it could be readily digested by restriction endonuclease. This method will aid rapid genetic classification of Parapoxvirus.
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Survey on antibody against Parapoxvirus among cattle in Japan.
Microbiology and immunology, 2000Co-Authors: Hiroshi Sentsui, Yasuo Inoshima, Kenji Murakami, Akihiro Minami, Yasunori Yamamoto, Shinya ShimizuAbstract:A seroepidemiological survey was performed on antibody against Parapoxvirus among cattle in Japan using the agar gel immunodiffusion test and enzyme-linked immunosorbent assay. A total 1,819 sera were collected from cattle in various parts of Japan for the survey. The positive rates were in the range of 40 to 98%, and the reactors increased gradually in number with advancing age. These results indicate that Parapoxvirus infection is already prevalent among cattle in Japan. It remains to be elucidated, however, whether the antibodies detected have been produced by infection of the same virus or other viruses that belong to the genus Parapoxvirus.
