Sero-epizootiological study of racehorses with pyrexia in the training centers of the Japan Racing Association.

Sugiura, Takeo; Matsumura, Tomio; Fukunaga, Yoshio; Hirasawa, Kazunori

doi:10.1292/jvms1939.49.1087

Cited by 25 publications

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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%

“…Thereafter both EHV-1 and -4 spread among various horse populations in Japan. Today, many racehorses at two Training Centers of the Japan Racing Association (JRA) have suffered from respiratory disease with pyrexia by EHV-1 or -4 infection [20]. 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].…”

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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.

et al. 1998

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“…Winter pyrexia caused by EHV-1 epizootics in the training centers of the Japan Racing Association is an important condition that needs to be controlled, because it can delay training schedules and cause horses to be scratched from races (13). The spread of EHV-1 in the training centers may be due to the fact that, in these centers, Ͼ2,000 racehorses are kept in close contact and are subjected to stress from cold weather and hard training.…”

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Successful Control of Winter Pyrexias Caused by Equine Herpesvirus Type 1 in Japanese Training Centers by Achieving High Vaccination Coverage

Bannai

Mae

Ode

et al. 2014

Clin. Vaccine Immunol.

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“…However, many EHV-4-infected racehorses in Japanese breeding and rearing farms have also become infected with EHV-1 in training centers during the winter. Such infection has caused fever and occasional disorders of the central nervous system (5,7). Therefore, it is important to understand the pathogenesis of EHV-4 to prevent infections with both EHV-1 and EHV-4.…”

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Development of an Equine Herpesvirus Type 4-Specific Enzyme-Linked Immunosorbent Assay Using a B-Cell Epitope as an Antigen

Maeda

Mizukoshi²,

Hamano³

et al. 2004

J Clin Microbiol

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Equine herpesvirus type 4 (EHV-4) causes a pyrogenic respiratory disease in young horses that have been bred and reared in farms (3), and EHV-1 is a major cause of respiratory disease among racehorses during the winter in Japan (5,7,8). Since these viruses are serologically cross-reactive, horses infected with EHV-4 have antibodies that function against EHV-1 and are thus somewhat protected from EHV-1 infection. However, many EHV-4-infected racehorses in Japanese breeding and rearing farms have also become infected with EHV-1 in training centers during the winter. Such infection has caused fever and occasional disorders of the central nervous system (5, 7). Therefore, it is important to understand the pathogenesis of EHV-4 to prevent infections with both EHV-1 and EHV-4.The C-terminal regions of EHV-1 and EHV-4 glycoprotein G (gG) are specifically recognized by sera from horses infected with the respective virus (2), and enzyme-linked immunosorbent assays (ELISAs) with type-specific regions as antigens can serologically distinguish infections with these viruses (1). A modification of a type-specific ELISA for Japanese isolates was useful for seroepizootic studies and for the serodiagnosis of infections with both types of EHV (8). This method did not detect any antibody induced by inactivated vaccine against EHV-1 but could distinguish between EHV-1 and EHV-4 infections in vaccinated and nonvaccinated horses (9). It was also reported that levels of immunoglobulin Ga (IgGa) antibody against the EHV-4-specific region increased as much as those of IgGb antibody after EHV-4 infection but decreased more rapidly than those of IgGb (6). Therefore, analysis of the an-* Corresponding author. Mailing address: Department of Veterinary Microbiology, Faculty of Agriculture, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan. Phone: 81-83-933-5887. Fax: 81-83-933-5887. E-mail: kmaeda@ yamaguchi-u.ac.jp. FIG. 1. Expression of the type-specific region and a stretch of the repeat sequence in the type-specific region of EHV-4 TH20p. (A) A type-specific region (GST-egG4P) and a repeat sequence (GSTegG4R) were expressed as GST fusion proteins in E. coli. Whole bacterial proteins were separated by SDS-PAGE and stained with Coomassie brilliant blue. (B) Whole bacterial proteins were separated by SDS-PAGE and transferred to polyvinylidene difluoride membranes. Proteins were detected with diluted serum from a horse experimentally infected with EHV-4 (horse 1) and peroxidase-conjugated anti-horse IgG. Molecular masses are shown at the left.

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Sero-epizootiological study of racehorses with pyrexia in the training centers of the Japan Racing Association.

Cited by 25 publications

References 0 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.

Successful Control of Winter Pyrexias Caused by Equine Herpesvirus Type 1 in Japanese Training Centers by Achieving High Vaccination Coverage

Development of an Equine Herpesvirus Type 4-Specific Enzyme-Linked Immunosorbent Assay Using a B-Cell Epitope as an Antigen

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