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[This corrects the article DOI: 10.3389/fimmu.2026.1764863.].

Objective: Equine herpes viruses (EHVs) are considered one of the most important respiratory pathogens in equids, resulting in serious outcomes for equine health worldwide. The objectives of the current research were the detection, molecular characterization, and isolation of EHV-1 and EHV-4 circulating within different equine populations in Egypt, either clinically or in apparently healthy horses. Material and Methods: A total of 120 field samples were collected, and DNA was extracted. Screening and typing of extracted DNA were done by consensus and conventional PCR assays for detection of EHV-1 and EHV-4, followed by sequencing and phylogenetic analysis to confirm the virus identity. Selected positive samples for both EHV-1 and EHV-4 were subjected to Madin-Darby bovine kidney (MDBK) cell lines for virus isolation. Results: The obtained results revealed that 58/120 (48%) samples were positive for EHVs. Typing of positive samples showed that EHV-1 was detected in (48/120) 40% of samples and EHV-4 was detected in (15/120) 12% of samples, while dual infection by both EHV-1 and 4 was detected in five samples. Conclusion: The current study revealed new data on the continuous circulation of EHV-1 and EHV-4 within equine populations in Egypt, and individual horses could be infected by multiple EHVs. In addition, latently infected horses are acting as potential reservoirs for frequent virus reactivation.

Abstract Background Equine herpes virus type 1 (EHV-1) infection in horses is associated with respiratory and neurologic disease, abortion, and neonatal death. Hypothesis Vaccines decrease the occurrence of clinical disease in EHV-1-infected horses. Methods A systematic review was performed searching multiple databases to identify relevant studies. Selection criteria were original peer-reviewed research reports that investigated the in vivo use of vaccines for the prevention of disease caused by EHV-1 in domesticated horses. Main outcomes of interest included pyrexia, abortion, neurologic disease, viremia, and nasal shedding. We evaluated risk of bias, conducted exploratory meta-analyses of incidence data for the main outcomes, and performed a GRADE evaluation of the quality of evidence for each vaccine subtype. Results A total of 1018 unique studies were identified, of which 35 met the inclusion criteria. Experimental studies accounted for 31/35 studies, with the remainder being observational studies. Eight vaccine subclasses were identified including commercial (modified-live, inactivated, mixed) and experimental (modified-live, inactivated, deletion mutant, DNA, recombinant). Risk of bias was generally moderate, often because of underreporting of research methods, and sample sizes were small leading to imprecision in the estimate of the effect size. Several studies reported either no benefit or minimal vaccine efficacy for the primary outcomes of interest. Meta-analyses revealed significant heterogeneity was present, and our confidence in the quality of evidence for most outcomes was low to moderate. Conclusions and Clinical Importance Our review indicates that commercial and experimental vaccines minimally reduce the incidence of clinical disease associated with EHV-1 infection.

Comparative transcriptomics offers a powerful approach to elucidate host–virus interactions across related pathogens, yet systematic evaluations across species-matched cellular systems remain limited. We performed a cross-species RNA sequencing analysis of respective species’ cells infected with three alphaherpesviruses—herpes simplex virus 1 (HSV-1), bovine alphaherpesvirus 1 (BHV-1), and equid alphaherpesvirus 1 (EHV-1)—to dissect conserved and virus-specific transcriptional responses. We show that certain orthologous genes and orthologous pathways are differentially regulated upon infection among the three species like pathways related to translation rRNA processing and TNF-alpha signalling. We find that the earliest sampled timepoint of infection, 2 h post infection (hpi), shows the most commonly enriched pathways among the three species compared to later timepoints. At 6 h and 9 h post infection, BHV-1- and EHV-1 infections have more in common with each other in terms of enriched pathways than with HSV-1 infections. Moreover, we provide a comprehensive analysis of temporal viral gene expression for all three herpesviruses. Together, these findings provide a comparative framework for understanding alphaherpevirus–host interactions and reveal both conserved core responses and species-specific transcriptional signatures. This work establishes a foundation for identifying broadly acting antiviral targets as well as virus-specific vulnerabilities that may inform host-directed therapies and cross-species disease management.

This study investigates the production of volatile organic compounds (VOCs) in RK-13 cells infected with three equine viruses representing different families: equine arteritis virus (EAV) ( ), equine herpesvirus 1 (EHV-1) ( ), and equine rhinitis B virus (ERBV) ( ). VOCs, which are byproducts of cellular metabolism and potential non-invasive diagnostic markers, were analyzed using headspace solid-phase microextraction (HS-SPME) and gas chromatography-mass spectrometry (GC-MS). Since viruses do not possess intrinsic metabolic activity, the observed changes in the VOC profiles were attributed to host responses, such as metabolic reprogramming, oxidative stress, and apoptosis. We hypothesized that each viral infection induces distinct metabolic changes, resulting in characteristic VOC signatures that mirror the virus type, replication kinetics, and cytopathic effects. Notably, viruses with rapid cytopathic effects (e.g., EHV-1) were anticipated to trigger more pronounced VOC alterations. In our experimental design, RK-13 cells were infected at a multiplicity of infection of 1 and incubated for 24 h, 48 h, or 72 h. Distinct VOC profiles emerged, with significant elevations in compounds like 2-ethyl-1-hexanol, particularly in EHV-1 infections, and selective increases in acetophenone and benzaldehyde. Principal component analysis (PCA) of the VOC concentration data showed the clear separation of data from viruses from different families. These findings support the potential of VOC profiling as a rapid diagnostic tool for viral infections.

The present study investigated the molecular detection and genetic characteristics of equine herpesvirus 1 (EHV‐1) and EHV‐4 circulating within Egyptian horse populations during 2019–2022. A total of 79 animals were sampled (54 nasal swabs and 25 aborted fetal tissues). PCR assays revealed that 24 (30.3%) and 7 (8.8%) samples were positive for EHV‐1 and EHV‐4, respectively. Additionally, 5 (6.3%) samples were concurrently infected with both viruses. Four EHV‐1 and three EHV‐4 isolates were genetically characterized based on partial sequencing of gB gene. The four EHV‐1 strains displayed 100% nucleotide identity to one another and to EHV‐1 reference strains reported in Egypt and other countries. The three EHV‐4 strains were phylogenetically classified into two distinct clusters based on their nucleotide sequences (76%–100% identity). Meanwhile, their deduced amino acid sequences differed by only one amino acid substitution. Our results underscore the critical importance of EHV‐1 and EHV‐4 as primary contributors of abortion and respiratory illness in horses and highlight the need for further large‐scale surveillance and in‐depth characterization studies to improve our understanding of these viruses’ epidemiology in Egypt and to develop a robust control strategy.

Equine herpesvirus type 1 (EHV-1) is a major veterinary pathogen causing significant economic losses in the livestock industry. Despite its impact, effective vaccines and targeted antiviral strategies remain limited, largely due to an incomplete understanding of host factors regulating viral replication and pathogenesis. To systematically identify host genes essential for EHV-1 infection, we established a BHK-21 cell line stably expressing Cas9 and performed a genome-wide CRISPR/Cas9 knockout screen using a pooled lentiviral single-guide RNA library. Significantly enriched candidate genes from positive selection were validated by generating knockout cell lines. Viral replication and protein expression were assessed using quantitative polymerase chain reaction and Western blot analysis. Pathway enrichment and protein interaction network analyses were subsequently conducted. Genome-wide CRISPR/Cas9 screening identified multiple host factors critical for EHV-1 replication. Pathway enrichment analysis revealed that these genes were involved in key cellular signaling and regulatory networks associated with viral infection. Functional validation demonstrated that knockout of selected host genes significantly suppressed EHV-1 replication and viral protein synthesis. These findings highlight essential host determinants required for EHV-1 replication and suggest that targeting host factors may represent a promising strategy for antiviral intervention. This study provides a foundation for the development of host-directed immunotherapeutic and antiviral approaches against EHV-1 infection.

Equine herpesvirus-1 is the cause of respiratory disease, abortion, and equine herpesvirus myeloencephalopathy (EHM) in horses worldwide. EHM affects as many as 14% of infected horses and a cell-associated viremia is thought to be central for EHM pathogenesis. While EHM is infrequent in younger horses, up to 70% of aged horses develop EHM. The aging immune system likely contributes to EHM pathogenesis; however, little is known about the host factors associated with clinical EHM. Here, we used the “old mare model” to induce EHM following EHV-1 infection. Peripheral blood mononuclear cells (PBMCs) of horses prior to infection and during viremia were collected and RNA sequencing with differential gene expression was used to compare the transcriptome of horses that did (EHM group) and did not (non-EHM group) develop clinical EHM. Interestingly, horses exhibiting EHM did not show respiratory disease, while non-EHM horses showed significant respiratory disease starting on day 2 post infection. Multiple immune pathways differed in EHM horses in response to EHV-1. These included an upregulation of IL-6 gene expression, a dysregulation of T-cell activation through AP-1 and responses skewed towards a T-helper 2 phenotype. Further, a dysregulation of coagulation and an upregulation of elements in the progesterone response were observed in EHM horses.

Varicellovirus equidalpha 1, formerly known as Equid alphaherpesvirus 1 (EHV-1), is highly prevalent and can lead to various problems, such as respiratory problems, abortion, neonatal foal death, and neurological disorders. The latter is known as equine herpes myeloencephalopathy (EHM). Cases of EHM have significantly increased since the beginning of the twenty-first century. The genomic sequences of five isolates associated with the fatal neurological outbreak in Valencia, Spain, in 2021 were analyzed and documented. The genome and replication kinetics of the Belgian EHM isolate 21P40, associated with the Valencia outbreak, and the well-characterized abortigenic strain 97P70 were compared. Both strains exhibited a nucleotide identity of 99.96%, with only seven genetic mutations in ORFs 13, 24, 30, 32, 40, 65, and 71. Isoleucine and asparagine at loci 291 and 207 of ORF30 (DNA polymerase) and ORF65 (ICP22), respectively, were unique to isolates from the Valencia outbreak. The replication kinetics of these two genetically closely related strains were determined in rabbit kidney (RK-13), equine respiratory, and vaginal mucosal explant cells, as well as equine blood monocytes (CD172a + ). Both strains replicated equally well in RK-13 cells. The neuropathogenic isolate 21P40 exhibited a more extensive infection in respiratory explants and blood monocytes, as demonstrated by more plaques and single infected leukocytes, and a higher percentage of infected monocytes. In contrast, vaginal explants infected with the abortigenic strain 97P70 demonstrated more plaques and single infected leukocytes. In conclusion, 21P40 replicated significantly different compared to 97P70 but shared similarities with the Belgian well-studied neuropathogenic EHV-1 strain 03P37.

Abstract Background Equine herpesvirus myeloencephalopathy (EHM) is a persistent threat to horses, with unclear risk factors and disease severity. Objectives To evaluate risk factors, effective reproduction rate (Rt), and long-term athletic outcomes of an EHM outbreak. Methods Retrospective study of the 2021 EHM outbreak in Valencia, Spain, examining associations between risk factors (sex, age, breed, country of origin, and vaccination status) and case fatality rate, EHM development, and odds of returning to competition using odds ratios [95% CI] and Rt via the Robert Kochs Institute method. Results Among 191 horses, 38 (20%) were clinically normal, 13 (7%) were subclinical, and 140 (73%) presented clinical signs (89 EHM, 64%). One hundred sixty horses were isolated at the show, while 47 were treated in hospitals. The mean age was 9.8 ± 3.0 years; 85 (45%) were mares, 79 (41%) geldings, and 27 (14%) stallions. The EHM case fatality rate was 11/89 (12%). Vaccination was associated with EHM development (4.54[2.23–9.27]; OR[95% CI]; p < 0.001) and case fatality rate (3.9[1.1–14.4]; OR[95% CI]; p < 0.043). EHV-1-infected horses without EHM were more likely to return to competition (54/61; 89%) than those recovering from EHM (65/89; 73%; p = 0.024). It was initially 4.2 and decreased to < 1 within 2 weeks of the outbreak. Conclusions During the 2021 EHV-1 outbreak in Valencia, vaccination status appears to be associated with EHM development. Horses recovering from EHM had slightly lower chances of returning to competition than those shedding EHV-1 without EHM. The high Rt value underscores the contagious nature of EHV-1.