Novel endotheliotropic herpesviruses fatal for Asian and African elephants

Science

Volumn 283, Issue 5405, 1999, Pages 1171-1176

  Richman, Laura K ^a   Montali, Richard J ^a   Garber, Richard L ^a   Kennedy, Melissa A ^a   Lehnhardt, John ^a   Hildebrandt, Thomas ^a   Schmitt, Dennis ^a   Hardy, Douglas ^a   Alcendor, Donald J ^a   Hayward, Gary S ^a  

^a NONE

Author keywords

[No Author keywords available]

Indexed keywords

ARTICLE; CLINICAL FEATURE; CROSS REACTION; DIAGNOSTIC APPROACH ROUTE; DNA SEQUENCE; ELEPHANT; HERPES; HERPES VIRUS; HISTOPATHOLOGY; NONHUMAN; NUCLEOTIDE SEQUENCE; PRIORITY JOURNAL; STRAIN DIFFERENCE; VIRUS ISOLATION;

AFRICA; AMINO ACID SEQUENCE; ANIMALS; ANIMALS, ZOO; ASIA; BASE SEQUENCE; DNA, VIRAL; DNA-DIRECTED DNA POLYMERASE; ELEPHANTS; ENDODEOXYRIBONUCLEASES; ENDOTHELIUM, VASCULAR; FEMALE; GENES, VIRAL; HEMORRHAGE; HERPESVIRIDAE; HERPESVIRIDAE INFECTIONS; INCLUSION BODIES, VIRAL; MALE; MOLECULAR SEQUENCE DATA; PHYLOGENY; POLYMERASE CHAIN REACTION; UNITED STATES; VIRAL PROTEINS;

ANIMALIA; ELEPHANT; ELEPHAS MAXIMUS; HERPES; HERPESVIRIDAE; LOXODONTA;

EID: 0033582783     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.283.5405.1171     Document Type: Article

References (15)

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2. L. K. Richman et al., in preparation.
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7. Initially, consensus primer PCR was performed using a published set of redundant primers for the terminase and DNA polymerase gene regions. Specific single primer sets for the elephant herpesvirus terminase and DNA polymerase gene regions were constructed from the sequence initially obtained from elephant cases 1 and 2: terminase primers for Asian elephants (5′-GTACGTCCTTTCTAGCTCAC-3′ and 5′-GTGTCG-GCTAAATGTTCTTG-3′), terminase primers for African elephants (5′-AATGTGATATCCTACGTATG-3′ and 5′-GTACTATATCTTATCATGTC-3′), and DNA polymerase primers for both elephant species (5′-GTGTCTGGCTATAGCAGAGT-3′ and 5′-CACATC-GATACGGAATCTCT-3′). PCR amplification was performed with nonredundant primers in a 50-μl reaction volume containing PCR SuperMix (Gibco/BRL-Life Technologies, Gaithersburg, MD), 0.3% (v/v) glycerol, and 20 pmol of each primer. Thirty-six cycles of PCR were completed using the following protocol: denaturation at 94°C for 1 min, annealing at 50°C for 1 min, and extension at 72°C for 2 min. The final extension was performed at 72°C for 7 min. The PCR product was visualized on a 1.5% agarose gel stained with ethidium bromide. A second round of PCR was needed to detect the African elephant vulval lymphoid patch product using the same terminase primers under identical conditions. Some PCR products were cloned using the TA Cloning kit (Invitrogen) and sequenced, and others were sequenced directly from the PCR products.
8. DNA was extracted from ∼100 μm of paraffin-embedded, formalin-fixed tissue with the Ex-wax kit (Oncor, Gaithersburg, MD). After precipitation, DNA was resuspended and amplified by PCR. DNA was extracted from 200 μl of whole blood (either fresh or frozen at -80°C) using Gentra System (Minneapolis, MN) capture columns according to the manufacturer's protocol.
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15. Funded by NIH grant No. 1 K08 Al01526-01, the Smithsonian Scholarly Studies Program, the Kumari Elephant Conservation Fund, and Friends of the National Zoo. We thank the following individuals and institutions for their contributions to our study: R. Viscidi, Johns Hopkins School of Medicine, for assistance with phylogenetic analysis; J. Strandberg, F. Hamzeh, and A. Shahkolahi, Johns Hopkins School of Medicine; J. Cohen, NIH; S. Feldman, Anmed/Biosafe Inc.; S. Mikota, Audubon Institute; R. Mirkovic, Southwest Foundation for Biomedical Research; J. d'Offay and R. Eberle, Oklahoma State University; G. Letchworth, University of Wisconsin-Madison; K. E. Steele, B. Connolly, and P. Jahrling, U.S. Army Medical Research Institute of Infectious Diseases; A. Ruebel, Zoo Zürich; P. Ossent, University of Zürich; F. Osorio, University of Nebraska, Lincoln; S. Kania, University of Tennessee, Knoxville; E. Dierenfeld, New York Wildlife Conservation Society; J. Trupkiewicz, L. Munson, and D. Taylor, University of California, Davis; D. Nichols, N. Spangler, K. Clark, J. Sutton, N. Pratt, J. Block, M. Bush, and the elephant keeper staff, National Zoological Park; J. Jenkins and R. V. Ferris, Armed Forces Institute of Pathology; J. Gaskin, University of Florida; D. Olson, African elephant Species Survival Plan (SSP) coordinator M. Keele, Asian elephant SSP coordinator; A. Schanberger, Houston Zoological Gardens; Marine World Africa-USA, Vallejo, CA; L. Bingaman-Lackey, AZA; N. Kriek, University of Pretoria, South Africa; R. Bengis, Kruger National Park, South Africa; and San Diego Zoo and Wild Animal Park, New York Wildlife Conservation Society, Lincoln Park Zoo, Dickerson Park Zoo, African Lion Safari, Tulsa Zoological Park, Fort Worth Zoo, Indianapolis Zoo, Dallas Zoo, Oakland Zoo, and the Center for Reproduction of Endangered Species.