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Detecting Herpesvirus By Negative Contrast Electon Microscopy in Tissue and Cell Culture From Mule Deer with Infectious Keratoconjunctivitis
Published online by Cambridge University Press: 02 July 2020
Abstract
Mule deer are infected with a herpesvirus associated with acute infectious keratoconjunctivitis (IKC) or “pink eye”. in December 2000, many young male mule deer in Wyoming had clinical signs of IKC including corneal edema, mucopuvulent conjuctivitis, blepharospasm, and blindness.
Conjunctival swab samples from 5 mule deer with IKC were processed for negative contrast electron microscopy (NCEM) at the Wyoming State Veterinary Laboratory (WSVL). Homogenized samples from each deer were centrifuged for 20 minutes at 3440g in a Beckman Avanti J-25 centrifuge (Beckman Instruments Inc., Fullerton, CA, USA). The supernatant from each sample was subsequently centrifuged for 60 minutes at 39,800g. The final pellet was resuspended in 210 μl of sterile deionized microfiltered water. 105 μ1 of the virus suspension was mixed with sodium phosphotungstic acid stain containing 84 μl of 4% aqueous phoshotungstic acid, pH 6.8 with NaOH, 420 μl sterile, deionized microfiltered water, and 21 μl 0.1% bovine serum albumin.
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- Microbiology
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- Copyright © Microscopy Society of America 2001