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Equine veterinary journal2023; doi: 10.1111/evj.14007

Detection of equine herpesvirus-1 (EHV-1) in urine samples during outbreaks of equine herpesvirus myeloencephalopathy.

Abstract: Real-time PCR is the diagnostic technique of choice for the diagnosis and control of equine herpesvirus-1 (EHV-1) in an outbreak setting. The presence of EHV-1 in nasal swabs (NS), whole blood, brain and spinal cord samples has been extensively described; however, there are no reports on the excretion of EHV-1 in urine, its DNA detection patterns, and the role of urine in viral spread during an outbreak. Objective: To determine the presence of EHV-1 DNA in urine during natural infection and to compare the DNA detection patterns of EHV-1 in urine, buffy coat (BC) and NS. Methods: Descriptive study of natural infection. Methods: Urine and whole blood/NS samples were collected at different time points during the hospitalisation of 21 horses involved in two EHV-1 myeloencephalopathy outbreaks in 2021 and 2023 in Spain. Quantitative real-time PCR was performed to compare the viral DNA load between BC-urine samples in 2021 and NS-urine samples in 2023. Sex, age, breed, presence of neurological signs, EHV-1 vaccination status and treatment data were recorded for all horses. Results: A total of 18 hospitalised horses during the 2021 and 2023 outbreaks were positive for EHV-1, and viral DNA was detected in urine samples from a total of 11 horses in both outbreaks. Compared with BC samples, DNA presence was detected in urine samples for longer duration and with slightly higher concentration; however, compared with NS, detection of EHV-1 in urine was similar in duration with lower DNA concentrations. Conclusions: Limited sample size, different sampling times and protocols (BC vs. NS) in two natural infection outbreak settings. Conclusions: EHV-1 was detected in the urine from naturally infected horses. Urine should be considered as complimentary to blood and NS in diagnosis of EHV-1 infection. Unassigned: PCR en tiempo real es la técnica diagnostica de preferencia para el diagnóstico y control del herpes virus equino-1 (EHV-1) en una situación de brote. La presencia de EHV-1 en torulas nasales (TN), muestras de sangre entera, cerebro, y medula espinal ha sido descrita en forma extensa; sin embargo, no hay informes de excreción de EHV-1 en orina, la detección del patrón de ADN, y el rol de la orina en la propagación vírica durante un brote. Objective: Determinar la presencia de ADN de EHV-1 en muestras de orina durante un brote infeccioso natural y comparar los patrones de detección de ADN de EHV-1 en orina, capa leucocitaria (CL) y TN. DISEÑO DEL ESTUDIO: Estudio prospectivo en una infección natural en caballos hospitalizados. MÉTODOS: Muestras de orina y sangre entera/TN fueron recolectadas a distintos tiempos durante la hospitalización de veintiún caballos involucrados en dos brotes de mielo encefalopatía por EHV-1 en 2021 y 2023 en España. PCR a tiempo real cuantitativo fue llevado a cabo para comparar la carga de ADN viral entre muestras de CL-orina en 2021 y muestras TN-orina en 2023. Sexo, edad, raza, presencia de síntomas neurológicos, estatus de vacunación y datos de tratamiento fueron anotados para todos los caballos. Results: Un total de diez y ocho caballos hospitalizados durante los brotes de 2021 y 2023 resultaron positivos a EHV-1, y ADN viral fue detectado en muestras de orina en un total de 11 caballos de ambos brotes. En comparación a muestras de CL, la presencia de AND fue detectado por mas largo tiempo y con una concentración ligeramente mas alta; sin embargo, en comparación a TN, la detección de EHV-1 en orina fue similar en tiempo pero demostró menor concentración de ADN. Unassigned: Tamaño de muestra limitado, tiempos de muestreo diferentes, y de protocolos (CL vs. TN) en dos situaciones de brotes naturales. Conclusions: Se detecto EHV-1 en orina de caballos infectados naturalmente. La recolección, no invasive, de orina debería considerarse como un complemento a las muestras de sangre y TN en el control de caballos infectados en situaciones de brote.
© 2023 The Authors. Equine Veterinary Journal published by John Wiley & Sons Ltd on behalf of EVJ Ltd.
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Publication Date: 2023-09-12 PubMed ID: 37699794DOI: 10.1111/evj.14007Google Scholar: Lookup
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Summary

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This research investigated the presence of equine herpesvirus-1 (EHV-1) in the urine of horses during outbreaks of equine herpesvirus myeloencephalopathy. Using real-time PCR techniques, the researchers discovered EHV-1 DNA in urine samples, pointing to urine as a crucial factor in viral spread during outbreaks.

Study Design

  • The researchers conducted a descriptive study during two natural outbreaks of EHV-1 myeloencephalopathy in Spain in 2021 and 2023.
  • They collected urine samples and whole blood/nasal swab samples from 21 hospitalized horses at different points in their treatment.
  • Details such as the horses’ sex, age, breed, neurological symptoms, vaccination status against EHV-1, and treatment data were all recorded.

Methodology

  • Quantitative real-time PCR (Polymerase Chain Reaction) was used to compare the viral DNA load between blood and urine samples in 2021, and nasal swab and urine samples in 2023.
  • PCR is a common method used in molecular biology to create many copies of a specific DNA segment, enabling precise detection and quantification of the target DNA.

Results

  • Out of the 21 sampled horses, 18 were positive for EHV-1 during both outbreaks.
  • EHV-1 DNA was detected in the urine samples from 11 of the 18 positive horses.
  • When compared to the blood samples, the presence of DNA was detected in urine samples for a more extended period and slightly higher concentration. However, urine and nasal swab samples showed similar duration of DNA presence, but urine samples had lower DNA concentrations.

Conclusions

  • The researchers noted limitations in their study, such as a relatively small sample size and different sampling times and protocols in the two natural infection outbreak settings.
  • However, they concluded that EHV-1 was indeed present in the urine of naturally infected horses during both studied outbreaks.
  • They also suggested that urine should be considered along with blood and nasal swabs during the diagnosis of EHV-1 infections in outbreak scenarios.

Cite This Article

APA
Velloso Alvarez A, Jose-Cunilleras E, Dorrego-Rodriguez A, Santiago-Llorente I, de la Cuesta-Torrado M, Troya-Portillo L, Rivera B, Vitale V, de Juan L, Cruz-Lopez F. (2023). Detection of equine herpesvirus-1 (EHV-1) in urine samples during outbreaks of equine herpesvirus myeloencephalopathy. Equine Vet J. https://doi.org/10.1111/evj.14007

Publication

Equine veterinary journal
ISSN: 2042-3306
NlmUniqueID: 0173320
Country: United States
Language: English

Researcher Affiliations

Velloso Alvarez, Ana
  • University Cardenal Herrera CEU, Valencia, Spain.
Jose-Cunilleras, E
  • Unitat Equina, Fundació Hospital Clínic Veterinari, Cerdañola del Valles, Spain.
  • Department of Animal Medicine and Surgery, Universitat Autònoma de Barcelona, Cerdañola del Valles, Spain.
Dorrego-Rodriguez, Abel
  • VISAVET Health Surveillance Centre, Universidad Complutense, Madrid, Spain.
Santiago-Llorente, Isabel
  • Hospital Clínico Veterinario Complutense, Universidad Complutense, Madrid, Spain.
de la Cuesta-Torrado, Maria
  • University Cardenal Herrera CEU, Valencia, Spain.
Troya-Portillo, Lucas
  • Unitat Equina, Fundació Hospital Clínic Veterinari, Cerdañola del Valles, Spain.
  • Department of Animal Medicine and Surgery, Universitat Autònoma de Barcelona, Cerdañola del Valles, Spain.
Rivera, Belen
  • VISAVET Health Surveillance Centre, Universidad Complutense, Madrid, Spain.
Vitale, Valentina
  • University Cardenal Herrera CEU, Valencia, Spain.
de Juan, Lucia
  • VISAVET Health Surveillance Centre, Universidad Complutense, Madrid, Spain.
  • Animal Health Department, Facultad de Veterinaria, Universidad Complutense, Madrid, Spain.
Cruz-Lopez, Fatima
  • VISAVET Health Surveillance Centre, Universidad Complutense, Madrid, Spain.

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Citations

This article has been cited 2 times.
  1. Musoles-Cuenca B, Padilla-Blanco M, Vitale V, Lorenzo-Bermejo T, de la Cuesta-Torrado M, Ballester B, Maiques E, Rubio-Guerri C, Velloso Alvarez A. First Molecular Evidence of Equine Herpesvirus Type 1 (EHV-1) in Ocular Swabs of Clinically Affected Horses. Viruses 2025 Jun 18;17(6).
    doi: 10.3390/v17060862pubmed: 40573453google scholar: lookup
  2. de la Cuesta-Torrado M, Velloso Alvarez A, Neira-Egea P, Cuervo-Arango J. Long-term performance of show-jumping horses and relationship with severity of ataxia and complications associated with myeloencephalopathy caused by equine herpes virus-1. J Vet Intern Med 2024 May-Jun;38(3):1799-1807.
    doi: 10.1111/jvim.17070pubmed: 38609161google scholar: lookup
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