Association of Two Newly Recognized Herpesviruses with Interstitial Pneumonia in Donkeys (<i>Equus Asinus</i>)

Kleiboeker, Steven B.; Schommer, Susan K.; Johnson, Philip J.; Ehlers, Bernhard; Turnquist, Susan E.; Boucher, Magalie; Kreeger, John M.

doi:10.1177/104063870201400401

Cited by 60 publications

(91 citation statements)

References 39 publications

(44 reference statements)

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“…128 Other investigators have successfully used this consensus PCR assay to identify novel herpesviruses. Kleiboeker et al 59 used FFPE lung tissues to investigate the presence of herpesvirus infections in donkeys with interstitial pneumonia. Williams et al 144 investigated whether a herpesvirus is associated with equine multinodular pulmonary fibrosis (EMPF), a newly described disease in horses.…”

Section: Herpesvirusesmentioning

confidence: 99%

Beyond H&E

et al. 2013

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Veterinary pathology of infectious, particularly viral, and neoplastic diseases has advanced significantly with the advent of newer molecular methodologies that can detect nucleic acid of infectious agents within microscopic lesions, differentiate neoplastic from nonneoplastic cells, or determine the suitability of a targeted therapy by detecting specific mutations in certain cancers. Polymerase chain reaction-based amplification of DNA or RNA and in situ hybridization are currently the most commonly used methods for nucleic acid detection. In contrast, the main methodology used for protein detection within microscopic lesions is immunohistochemistry. Other methods that allow for analysis of nucleic acids within a particular cell type or individual cells, such as laser capture microdissection, are also available in some laboratories. This review gives an overview of the factors that influence the accurate analysis of nucleic acids in formalin-fixed tissues, as well as of different approaches to detect such targets. Keywords cancer, ISH, IHC, nucleic acid analysis, molecular pathology, PCR, virology Technical ConsiderationsTissue Handling, Fixation, and Embedding Fixation of tissues in 10% neutral buffered formalin, followed by paraffin embedding (FFPE), has long been the standard approach of tissue preparation for microscopic evaluation. The chemical principle of formalin fixation is based on the crosslinking of proteins. Formalin has been used extensively because it is inexpensive and excellently preserves morphological detail in histological sections. However, the relatively recent development and increasing use of both protein and nucleic acid detection methods have revealed some significant shortcomings associated with the formalin-based fixation process. The effects of formalin fixation on DNA are chemical modification, DNA trapping, and potentially DNA fragmentation. Chemical modification, considered the main effect, results from the direct cross-linking of proteins to DNA. 68 In addition, extensive protein-protein cross-linking and the formation of protein networks result in DNA trapping. The combination of these effects makes it much more difficult to extract DNA from FFPE tissue than from unfixed tissue, especially when methods used for extraction from fresh tissues are used on FFPE tissue without any modifications. 60,83 In addition to interfering with the ability to extract nucleic acid, formalin fixation also causes DNA fragmentation, especially when the formalin used is inadequately buffered, which then leads to formic acid formation during the fixation process. This, in turn, results in depurination of the DNA, which then becomes susceptible to cleavage by hydroxyl ions and results in the formation of single-stranded breaks.60 Similar to DNA, RNA is also degraded and chemically modified as the result of formalin fixation. The most important chemical effect of formalin on RNA is the formation of monomethylol groups with purine bases. 77 This has a significant effect on reverse transcription ...

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Section: Herpesvirusesmentioning

confidence: 99%

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et al. 2013

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“…Gammaherpesviruses have also been detected in donkeys. 5,21 The first gammaherpesvirus of donkeys to be detected, designated asinine herpesvirus 2 (AsHV2), was isolated from the blood of a healthy donkey. 5 In contrast, asinine herpesviruses 4 (AsHV4) and asinine herpesvirus 5 (AsHV5), both of which are most similar genomically to gammaherpesviruses of equids, were detected in lung tissue of donkeys in which acute and typically fatal respiratory disease was a prominent feature of the clinical history.…”

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Detection and Nucleotide Sequencing of a DNA-Packaging Protein Gene of Equine Gammaherpesviruses

et al. 2004

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“…3,6,9 Asinine herpesviruses 4, 5, and 6 were designated as novel gammaherpesviruses based on sequences obtained from donkeys with interstitial pneumonia. 7,8 The latter viruses were highly similar over ,165 bp in the DNA polymerase gene and ,380 bp in the terminase gene and appeared to represent closely related gammaherpesviruses. Overall, paucity of sequence information for AsHV-1-3, and limited sequence availability for AsHV-4-6, constrains comparison and classification of equine herpesviruses.…”

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confidence: 99%

“…5 Asinine herpesviruses previously identified include AsHV-1 associated with coital exanthema, AsHV-2 isolated from leukocytes of healthy animals, AsHV-3 obtained from the nasal cavity of immunosuppressed animals, 2 and AsHV-4, -5, and -6 from animals with interstitial pneumonia. 7,8 Asinine herpesvirus 1 was classified as an alphaherpesvirus, but had little similarity with EHV-3 by restriction endonuclease and Southern blot analysis.…”

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confidence: 99%

“…These sequences had previously been identified in donkeys with interstitial pneumonia, and comparison with other herpesviruses grouped them into the subfamily Gammaherpesvirinae. 7 To further investigate the presence of a herpesvirus in this donkey with neurological disease, a second deep pharyngeal swab was obtained, and DNA was extracted again. Since longer sequences of asinine herpesviruses were not available in the NCBI database, polymerase gene sequences of EHV-1, -2, -4, -5, and -7 were aligned, f and conserved regions were chosen for primer design.…”

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Herpesvirus-Associated Neurological Disease in a Donkey

2008

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Abstract. A 4-year-old donkey was evaluated for progressive neurological abnormalities consisting of depression, stupor, weakness, and recumbency. Diagnostic evaluation for viral involvement identified an asinine herpesvirus in DNA extracted from deep pharyngeal swabs. Specific primers were designed based on comparison with equine herpesviral DNA polymerase sequences and yielded an 875-base pair product from the donkey. This sequence had complete identity with short sequences of asinine herpesvirus previously identified in donkeys with interstitial pneumonia. Amino acid analysis of the entire sequence indicated high similarity with Equid herpesvirus 7 (91%), Zebra herpesvirus 1 (90%), and Equid herpesvirus 2 (89%). With supportive treatment and physical therapy, the donkey gradually recovered over 5 days of hospitalization and returned to normal function. The current case illustrates the potential of a novel asinine herpesvirus to induce neurological disease in donkeys and provides a large viral sequence allowing confident assignment of this virus to the subfamily Gammaherpesvirinae.

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Association of Two Newly Recognized Herpesviruses with Interstitial Pneumonia in Donkeys (Equus Asinus)

Cited by 60 publications

References 39 publications

Beyond H&E

Beyond H&E

Detection and Nucleotide Sequencing of a DNA-Packaging Protein Gene of Equine Gammaherpesviruses

Herpesvirus-Associated Neurological Disease in a Donkey

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