Outbreak of equine herpesvirus 4 (EHV-4) in Denmark: tracing patient zero and viral characterization.
Abstract: Equine herpesvirus 4 (EHV-4) causes respiratory disease in horses, and the virus is considered endemic in the global equine population. However, outbreaks can occur when several horses are gathered in relation to shows, competitions, breeding units and at hospitals. In the spring year 2022, an EHV-4 outbreak occurred at the Large Animal Teaching Hospital, University of Copenhagen, Denmark. Nine horses were tested EHV-4 positive during the outbreak, which lasted approx. seven weeks. In addition, a tenth horse "Eq10" tested EHV-4 positive almost three weeks after the last of the outbreak horses tested positive. Detailed clinical registrations were obtained from all ten horses as well as their location and movement during hospitalization. Nasal swabs were obtained throughout the outbreak and tested by qPCR for EHV-4. Additionally, pre- and post-infection sera were tested for the presence of EHV-4 antibodies. Selected samples were characterized by partial and full genome sequencing. Results: The most common clinical signs of the EHV-4 infected horses during this outbreak were pyrexia, nasal discharge, mandibular lymphadenopathy and increased lung sounds upon auscultation. Based on the locations of the horses, EHV-4 detection and antibody responses the most likely "patient zero" was identified as being "Eq1". Partial genome sequencing revealed that Eq10 was infected by another wild type EHV-4 strain, suggesting that the hospital was able to eliminate the outbreak by testing and reinforcing biosecurity measures. The complete genome sequence of the outbreak strain was obtained and revealed a closer relation to Australian and Japanese EHV-4 strains rather than to other European EHV-4 strains, however, very limited sequence data are available from Europe. Conclusions: The study illustrated the transmission of EHV-4 within an equine facility/hospital and provided new insights into the viral shedding, antibody responses and clinical signs related to EHV-4 infections. Finally, sequencing proved a useful tool in understanding the transmission within the hospital, and in characterizing of the outbreak strain.
© 2024. The Author(s).
Publication Date: 2024-07-03 PubMed ID: 38961400PubMed Central: PMC11221098DOI: 10.1186/s12917-024-04149-xGoogle Scholar: Lookup
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- Journal Article
Summary
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Overview
- This research examines an outbreak of equine herpesvirus 4 (EHV-4) at a Danish veterinary hospital in 2022, identifying the initial infected horse and characterizing the virus through clinical, molecular, and genomic analyses.
Background and Context
- EHV-4 is a virus that causes respiratory illness in horses worldwide and is commonly present (endemic) in horse populations.
- Outbreaks typically occur when horses congregate in settings such as shows, breeding centers, competitions, and hospitals.
- The study focuses on an outbreak at the Large Animal Teaching Hospital at the University of Copenhagen, Denmark, in spring 2022.
Outbreak Description and Sample Collection
- The outbreak lasted approximately seven weeks and initially involved nine horses testing positive for EHV-4 via nasal swab qPCR testing.
- A tenth horse (“Eq10”) tested positive about three weeks after the initial group’s last positive case, raising questions about ongoing transmission.
- Clinical data were recorded for all ten horses, including symptoms and movement within the hospital.
- Repeated nasal swab samples were taken during the outbreak to monitor viral presence and shedding using quantitative PCR.
- Blood samples were collected before and after infection to measure antibody responses to EHV-4, providing insight into immunity development.
Clinical Findings
- The infected horses commonly showed:
- Pyrexia (fever)
- Nasal discharge
- Mandibular lymphadenopathy (swollen lymph nodes under the jaw)
- Increased lung sounds on auscultation, indicating respiratory involvement
- These symptoms are consistent with respiratory disease caused by EHV-4.
Tracing and Identification of Patient Zero
- By correlating horse locations, timing of EHV-4 detection, and antibody responses, the researchers identified horse “Eq1” as the probable source of the outbreak (“patient zero”).
Viral Characterization and Whole Genome Sequencing
- Partial genome sequencing of the virus from horse Eq10 revealed infection by a different EHV-4 strain compared to the outbreak strain affecting the initial nine horses.
- This suggests that the hospital successfully contained the outbreak through testing and enhanced biosecurity measures, preventing the spread of the original virus strain.
- The researchers sequenced the complete genome of the outbreak strain, revealing:
- Close genetic similarity to EHV-4 strains from Australia and Japan.
- A distant relationship to other European EHV-4 strains, but with limited sequence data available from Europe, making comprehensive comparisons difficult.
Conclusions and Implications
- The study demonstrates how EHV-4 can spread within a veterinary hospital setting and highlights the value of detailed clinical monitoring combined with molecular diagnostics.
- Sequencing played a critical role in:
- Identifying transmission chains during the outbreak.
- Differentiating new cases due to different viral strains from the initial outbreak cases.
- Characterizing the genetic makeup of the circulating virus.
- Findings from this study can inform future outbreak control strategies in equine facilities by emphasizing the importance of testing, biosecurity, and molecular surveillance.
Cite This Article
APA
Ryt-Hansen P, Johansen VK, Cuicani MM, Larsen LE, Hansen S.
(2024).
Outbreak of equine herpesvirus 4 (EHV-4) in Denmark: tracing patient zero and viral characterization.
BMC Vet Res, 20(1), 287.
https://doi.org/10.1186/s12917-024-04149-x Publication
Researcher Affiliations
- Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Grønnegårdsvej 2, Frederiksberg C, DK-1870, Denmark. piarh@sund.ku.dk.
- Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Grønnegårdsvej 2, Frederiksberg C, DK-1870, Denmark.
- Statens Serum Institut, Artillerivej 5, Copenhagen S, 2300, Denmark.
- Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Grønnegårdsvej 2, Frederiksberg C, DK-1870, Denmark.
- Department of Veterinary Clinical Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Taastrup, Denmark.
MeSH Terms
- Animals
- Horses
- Horse Diseases / virology
- Horse Diseases / epidemiology
- Disease Outbreaks / veterinary
- Denmark / epidemiology
- Herpesviridae Infections / veterinary
- Herpesviridae Infections / epidemiology
- Herpesviridae Infections / virology
- Herpesvirus 4, Equid / isolation & purification
- Male
- Female
- Antibodies, Viral / blood
- Hospitals, Animal
Conflict of Interest Statement
The authors declare no competing interests.
References
This article includes 28 references
- Azab W, Kato K, Abdel-Gawad A, Tohya Y, Akashi H. Equine herpesvirus 4: recent advances using BAC technology. Vet Microbiol. 2011;150(1–2):1–14. doi: 10.1016/j.vetmic.2011.01.002.
- Badenhorst M, Page P, Ganswindt A, Laver P, Guthrie A, Schulman M. Detection of equine herpesvirus-4 and physiological stress patterns in young thoroughbreds consigned to a South African auction sale. BMC Vet Res. 2015 [cited 2023 Mar 29];11(1). /pmc/articles/PMC4450643/
- Vaz PK, Horsington J, Hartley CA, Browning GF, Ficorilli NP, Studdert MJ et al. Evidence of widespread natural recombination among field isolates of equine herpesvirus 4 but not among field isolates of equine herpesvirus 1. J Gen Virol. 2016 [cited 2023 Mar 29];97(Pt 3):747. /pmc/articles/PMC5381393/
- Pavulraj S, Eschke K, Theisen J, Westhoff S, Reimers G, Andreotti S et al. Equine herpesvirus type 4 (Ehv-4) outbreak in germany: virological, serological, and molecular investigations. Pathogens. 2021 [cited 2023 Mar 29];10(7). /pmc/articles/PMC8308676/
- Izume S, Kirisawa R, Ohya K, Ohnuma A, Kimura T, Omatsu T et al. The full genome sequences of 8 equine herpesvirus type 4 isolates from horses in Japan. J Vet Med Sci. 2017 [cited 2023 Mar 29];79(1):206. /pmc/articles/PMC5289262/
- Pusterla N, James K, Barnum S, Bain F, Barnett DC, Chappell D et al. Frequency of detection and prevalence factors associated with common respiratory pathogens in equids with acute onset of fever and/or respiratory signs (2008–2021). Pathogens. 2022 [cited 2023 Mar 29];11(7). /pmc/articles/PMC9317490/
- Ma G, Azab W, Osterrieder N. Equine herpesviruses type 1 (EHV-1) and 4 (EHV-4)--masters of co-evolution and a constant threat to equids and beyond. Vet Microbiol. 2013 [cited 2023 Mar 29];167(1–2):123–34. https://pubmed.ncbi.nlm.nih.gov/23890672/
- Pusterla N, Leutenegger CM, Wilson WD, Watson JL, Ferraro GL, Madigan JE. Equine herpesvirus-4 kinetics in peripheral blood leukocytes and nasopharyngeal secretions in foals using quantitative real-time TaqMan PCR. J Vet Diagn Invest. 2005 [cited 2023 Mar 29];17(6):578–81. https://pubmed.ncbi.nlm.nih.gov/16475518/
- El-Hage C, Mekuria Z, Dynon K, Hartley C, McBride K, Gilkerson J. Association of equine herpesvirus 5 with mild respiratory disease in a survey of EHV1, -2, -4 and – 5 in 407 Australian horses. Anim an open access J from MDPI. 2021 [cited 2023 Mar 29];11(12). https://pubmed.ncbi.nlm.nih.gov/34944194/
- Pusterla N, Mapes S, David Wilson W. Prevalence of latent alpha-herpesviruses in thoroughbred racing horses. Vet J. 2012 [cited 2023 Mar 29];193(2):579–82. https://pubmed.ncbi.nlm.nih.gov/22405721/
- Cuxson JL, Hartley CA, Ficorilli NP, Symes SJ, Devlin JM, Gilkerson JR. Comparing the genetic diversity of ORF30 of Australian isolates of 3 equid alphaherpesviruses. Vet Microbiol. 2014;169(1–2):50–7. doi: 10.1016/j.vetmic.2013.12.007.
- Radalj A, Milic N, Stevanovic O, Nisavic J. The detection and phylogenetic analysis of equine herpesviruses 1, 4 and 5 identified in nasal swab samples of asymptomatic horses from Serbia and Bosnia and Herzegovina. Vet Ital. 2021 [cited 2023 Mar 29];57(4):265–74. https://pubmed.ncbi.nlm.nih.gov/35593499/
- Pusterla N, Bain F, James K, Mapes S, Kenelty K, Barnett DC et al. Frequency of molecular detection of equine herpesvirus-4 in nasal secretions of 3028 horses with upper airway infection. Vet Rec. 2017 [cited 2023 Apr 3];180(24):593. https://pubmed.ncbi.nlm.nih.gov/28386031/
- Sellon DC, Long MT. Equine infectious diseases. 2013.
- Kydd JH, Townsend HGG, Hannant D. The equine immune response to equine herpesvirus-1: the virus and its vaccines. [cited 2023 Sep 13]; Available from: www.elsevier.com/locate/vetimm
- QIAGEN. QIAamp DNA mini and blood mini handbook. 2016. p. 72. https://www.qiagen.com/us/resources/resourcedetail?id=62a200d6-faf4-469b-b50f-2b59cf738962&lang=en
- Indical bioscience, Handbook, SVANOVIR EHV1/EHV4-Ab. 2021. https://shop.indical.com/index.php?cl=details&anid=2b05cf3ab758820ec001dd7d8813152b&force_admin_sid=o5pe7kkq49cvinffcq4tqhorg3&stoken=93014DE9&shp=1&preview=e7d04d86fd89153f68a4a3f88edf4b41
- Chen S, Zhou Y, Chen Y, Gu J. fastp: an ultra-fast all-in-one FASTQ preprocessor. Bioinformatics. 2018 [cited 2023 Aug 4];34(17):i884–90. 10.1093/bioinformatics/bty560
- Pusterla N, Rice M, Henry T, Barnum S, James K. Investigation of the shedding of selected respiratory pathogens in healthy horses presented for routine dental care. 10.1177/0898756420949135. 2020 [cited 2023 Apr 4];37(2):88–93. https://journals.sagepub.com/doi/10.1177/0898756420949135
- AAEP. General biosecurity guidelines. Guideline. 2022 [cited 2022 Apr 1]. pp. 1–17. https://aaep.org/document/general-biosecurity-guidelines
- Li H, Durbin R. Fast and accurate short read alignment with Burrows-Wheeler transform. Bioinformatics. 2009 [cited 2023 Aug 4];25(14):1754–60. https://pubmed.ncbi.nlm.nih.gov/19451168/
- Li H. Aligning sequence reads, clone sequences and assembly contigs with BWA-MEM. 2013 [cited 2023 Aug 4]; https://arxiv.org/abs/1303.3997v2
- Danecek P, Bonfield JK, Liddle J, Marshall J, Ohan V, Pollard MO et al. Twelve years of SAMtools and BCFtools. Gigascience. 2021 [cited 2023 Aug 4];10(2):1–4. 10.1093/gigascience/giab008
- Katoh K, Standley DM. MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Mol Biol Evol. 2013 [cited 2023 Aug 4];30(4):772. /pmc/articles/PMC3603318/
- Capella-Gutiérrez S, Silla-Martínez JM, Gabaldón T. trimAl: a tool for automated alignment trimming in large-scale phylogenetic analyses. Bioinformatics. 2009 [cited 2023 Aug 4];25(15):1972. /pmc/articles/PMC2712344/
- Nguyen LT, Schmidt HA, Von Haeseler A, Minh BQ. IQ-TREE: a fast and effective stochastic algorithm for estimating maximum-likelihood phylogenies. Mol Biol Evol. 2015 [cited 2023 Aug 4];32(1):268–74. 10.1093/molbev/msu300
- Kalyaanamoorthy S, Minh BQ, Wong TKF, Von Haeseler A, Jermiin LS. ModelFinder: fast model selection for accurate phylogenetic estimates. Nat Methods 2017 146. 2017 [cited 2023 Aug 4];14(6):587–9. https://www.nature.com/articles/nmeth.4285
- Edgar CR. MUSCLE: multiple sequence alignment with high accuracy and high throughput. 2013 [cited 2019 Jun 27]; https://findit.dtu.dk/en/catalog/2354275506
Citations
This article has been cited 2 times.- Tallmadge RL, Laverack M, Lejeune M, Crossley B, Diel DG. A multiplex real-time PCR assay for detection of equid herpesvirus 1 and 4.. Sci Rep 2025 Oct 31;15(1):38201.
- Kutumbetov L, Myrzakhmetova B, Tussipova A, Zhapparova G, Tlenchiyeva T, Bissenbayeva K, Nurabayev S, Kerimbayev A. Development and Preclinical Evaluation of a Lyophilized Vaccine Against Equine Herpesvirus Type 4 (EHV-4).. Vaccines (Basel) 2025 May 31;13(6).
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