Epizootiological Aspects of Type 1 and Type 4 Equine Herpesvirus Infections among Horse Populations.

Matsumura, Tomio; Sugiura, Takeo; Imagawa, Hiroshi; Fukunaga, Yoshio; Kamada, Masanobu

doi:10.1292/jvms.54.207

Cited by 112 publications

(99 citation statements)

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“…ELISA titers against EHV-1 gG significantly rose in 30 (37.5%) out of 80 horses and those against EHV-4 gG rose in 9 (11.25%). These results were consistent with previous reports that EHV-1 is responsible for epizootic respiratory diseases in racehorses in winter [10,20], and revealed that EHV-4 is also sporadically responsible for respiratory disease among racehorses during the period. In addition, sera of two horses (Table 2, M28 and M42) reacted to both antigens.…”

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confidence: 93%

“…Furthermore, foals and yearlings suffering from respiratory disease on breeding and rearing farms also have been diagnosed serologically to be infected with EHV-1 or -4 [8,19]. The virological study revealed that EHV-1 caused epizootics of respiratory disease among racehorses exclusively in the winter season, and that EHV-4 also caused respiratory disease among racehorses sporadically and among foals and rearing horses irrespective of the season [10].…”

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confidence: 99%

“…[1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16] in which a tentative serological diagnosis of EHV-1 or -4 infection was made by complement fixation (CF) test. The result showed that a final diagnosis of EHV-1 infection was made in 8 horses and that diagnosis of EHV-1 infection might be made in another 3 horses (Table 1).…”

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Diagnosis and Sero-Epizootiology of Equine Herpesvirus Type 1 and Type 4 Infections in Japan Using a Type-Specific ELISA.

et al. 1998

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ABSTRACT. Recently, a type-specific ELISA using equine herpesvirus type 1 (EHV-1) and type 4 (EHV-4) glycoprotein Gs (gGs) was developed by Crabb and Studdert [1993]. To investigate the dissemination of EHV-1 and -4 among horses in Japan, we applied their ELISA as suitable for discriminating between EHV-1 and -4 infections serologically. Type-specificity of the ELISA was confirmed by using paired sera of infected horses with either EHV-1 or -4. Application of the ELISA to sera collected before and after the winter season of 1995-1996 from 80 racehorses revealed that 30 horses showed significant antibody responses against EHV-1 and 9 against EHV-4, respectively. The results indicated that this ELISA using paired sera is useful for a diagnosis and an epizootiological study on EHV-1 and -4 infections. -KEY WORDS: diagnosis, epizootiology, equine herpesvirus.J. Vet. Med. Sci. 60(10): 1133-1137, 1998 Virological diagnosis can identify the type of infected virus, but they are time-consuming and laborious. Therefore, it is considered to be very important to diagnose the type of infected virus serologically. The purpose of this study was to develop a type-specific serological test for diagnosis of EHV-1 and -4 infections, which facilitates the elucidation of epizootiological characteristics of both infections among the various horse populations in Japan.The EHV-1 89c25p strain [9] and an EHV-4 strain TH20, the Japanese prototype of EHV-4 [4], were used in this study. Viruses were propagated at low multiplicities of infection in primary fetal horse kidney cells. Infected cells showing a cytopathic effect were treated with 1% sodium dodecyl sulfate (SDS) and proteinase K (0.1 mg/ml) in 0.1 M Tris-HCl (pH 8.0), 0.1M NaCl, 5 mM EDTA at 37°C overnight. Following the DNA extraction with phenol and chloroform-isoamyl alcohol (24:1), ethanol precipitated preparations were dissolved in water.Crabb and Studdert [3] reported that type-specific, continuous epitopes were located in the corresponding Cterminal variable region of EHV-1 gG (amino acids 288 to 350) and EHV-4 gG (amino acids 287 to 382), which show little amino acid identity. To express the type-specific antigen, the bacterial expression plasmid pGEX-5X1 was used. The recombinant pGEX-5X1 plasmids, pGEX-gG1P and pGEX-gG4P, were constructed according to the methods described previously [3] with a minor modification. (i) pGEX-gG1P was constructed by PCR amplification (35 cycles of 92°C for 1 min, 55°C for 1 min, 72°C for 1 min) with the EHV-1 89c25p DNA as the template and the following primers: forward primer, gG1P-F, containing an EcoRI site, 5'-CCGAATTCGGTGACGAAACATACGA-3', and reverse primer, gG1P-R, containing an XhoI site, 5'-AACTCGAGCTGGATGCCGTTCGACG-3'. The amplified fragment was digested with EcoRI and XhoI and cloned into the pGEX-5X1 digested with the same restriction endonucleases. (ii) pGEX-gG4P was constructed by PCR amplification (35 cycles of 92°C for 1 min, 50°C for 1 min,

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Diagnosis and Sero-Epizootiology of Equine Herpesvirus Type 1 and Type 4 Infections in Japan Using a Type-Specific ELISA.

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“…The titration of viruses was conducted by plaque formation using MDBK cells and methylcellulose medium for overlay [17]. The reference viruses used were 89c25 for EHV-1 [18,19] and TH-20 for .Two half-bred weanling horses aged 8 months (H1 and H2) were used for inoculation of EHV-9. Ten ml of virus solution containing 10 7 plaque forming units (pfu) were inoculated intranasally using a nebulizer as described previously [17].…”

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Pathogenicity of a New Neurotropic Equine Herpesvirus 9(Gazelle Herpesvirus 1) in Horses.

Taniguchi

Fukushi

Matsumura

et al. 2000

The Journal of Veterinary Medical Science

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ABSTRACT. Pathogenicity of equine herpesvirus 9 (EHV-9), a new type of equine herpesvirus isolated from Gazella thomsoni, in horses was investigated by intranasal inoculation of EHV-9 (10 7 pfu) to two conventionally reared 8-months old half-bred weanling horses. Fever higher than 39°C was recorded. Virus was recovered from nasal swabs and peripheral blood mononuclear cells. Both horses developed neutralizing antibody to EHV-9. Perivascular infiltration of mononuclear cells and glial reaction were found in the olfactory and limbic systems. The results suggested that EHV-9 has a pathogenicity in horses.-KEY WORDS: equine, gazelle, herpesvirus.J. Vet. Med. Sci. 62(2): 215-218, 2000 describe the experimental infection of horses with EHV-9. The working seed of EHV-9 P19 [11] was prepared by propagation in fetal horse kidney (FHK) cells from the original stock of the third passage in Madin-Darby bovine kidney (MDBK) cells. The titration of viruses was conducted by plaque formation using MDBK cells and methylcellulose medium for overlay [17]. The reference viruses used were 89c25 for EHV-1 [18,19] and TH-20 for .Two half-bred weanling horses aged 8 months (H1 and H2) were used for inoculation of EHV-9. Ten ml of virus solution containing 10 7 plaque forming units (pfu) were inoculated intranasally using a nebulizer as described previously [17]. The inoculated horses were kept in a P3-level specific pathogen free facility of the Japan Racing Association, Tochigi, Japan.Rectal temperatures were recorded twice a day throughout the experiment. Fever higher than 39°C was recorded at 2, 4 and 5, and 1, 6 and 8 days post infection in H1 and H2, respectively. Other clinical symptoms were not observed. Leukocyte counts of both horses were normal during the experiment. Nasal swab and peripheral blood samples were collected daily for the first week and every other day for the second week after inoculation in order to investigate virus secretion and viremia. Viral isolation was conducted immediately after sample collection without freezing as described elsewhere [17]. A nasal swab was suspended in 2.0 ml phosphate buffered saline containing antibiotics. Approximately 10 6 mononuclear cells fractionated using Lymphoprep (Nycomed, Oslo, Norway) and 0.5 ml of nasal suspension were co-cultured with FHK cells in 25-cm 2 flasks (Nalge Nunc International Inc., Tokyo, Japan). The flasks were incubated at 37°C and observed for cytopathic effect for a week. Virus was recovered from nasal swabs from both horses on days 1 to 4, and from peripheral blood mononuclear cells in horse H1 on days 1 and 3 to 6 post infection. The recovered virus was identified as EHV-9 by A new herpesvirus was isolated from the epizootic encephalitis of Thomson's gazelles (Gazella thomsoni) kept at a zoological garden [11,25]. The outbreak occurred in a herd of ten gazelles in the fall of 1993. Seven gazelles died with or without apparent neurological symptoms. All dead gazelles had nonsuppurative encephalitis characterized by necrosis and degeneration of neuron...

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An Equine Herpesvirus Type 1 Recombinant with a Deletion in the gE and gI Genes Is Avirulent in Young Horses

et al. 1998

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The cell culture-adapted KyA strain of equine herpesvirus type 1 (EHV-1) has been found to be attenuated in young horses (Matsumura et al., 1996, Vet. Microbiol. 48, 353-365). The KyA strain lacks at least six genes in its genome, including those encoding glycoproteins gE and gI. To elucidate whether EHV-1 glycoproteins gE and gI play a role in viral virulence, we have constructed an EHV-1 recombinant that has the genes encoding both gE and gI deleted from its genome and its revertant. Growth properties of the deletion mutant virus in vitro were compared with those of the parent and the revertant viruses. Plaque size of the mutant virus in fetal horse kidney (FHK) cells was significantly smaller than those of the parent and the revertant viruses. In one-step growth experiments, however, the yields of infectious virus from FHK cells infected with the deletion mutant, the parent, or the revertant virus were approximately the same. The results suggested that gE and/or gI of EHV-1 promoted cell-to-cell spread of the virus, but that these glycoproteins were not involved in the process of virus maturation and release or in virus attachment and penetration. Subsequently, the virulence of mutant and revertant viruses was examined in young horses. No clinical signs were observed in six horses, including three colostrum-deprived foals inoculated intranasally with the deletion mutant virus, whereas three colostrum-deprived foals inoculated intranasally with the revertant virus manifested clinical signs typical for EHV-1 respiratory infection (i.e., pyrexia, nasal discharge, and swelling of submandibular lymph nodes). The results obtained from in vivo studies revealed that the EHV-1 mutant defective in both gE and gI genes was avirulent in young horses, suggesting that gE and/or gI of the EHV-1 have an important role in EHV-1 virulence. However, the EHV-1 mutant defective in both gE and gI genes induced only a partial protectivity in inoculated foals from manifestation of respiratory symptoms after challenge infection.

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Epizootiological Aspects of Type 1 and Type 4 Equine Herpesvirus Infections among Horse Populations.

Cited by 112 publications

References 13 publications

Diagnosis and Sero-Epizootiology of Equine Herpesvirus Type 1 and Type 4 Infections in Japan Using a Type-Specific ELISA.

Diagnosis and Sero-Epizootiology of Equine Herpesvirus Type 1 and Type 4 Infections in Japan Using a Type-Specific ELISA.

Pathogenicity of a New Neurotropic Equine Herpesvirus 9(Gazelle Herpesvirus 1) in Horses.

An Equine Herpesvirus Type 1 Recombinant with a Deletion in the gE and gI Genes Is Avirulent in Young Horses

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