Rapid subtyping of equine herpesvirus 1 with monoclonal antibodies.

Two antigenically similar subtypes of equine herpesvirus 1 (EHV-1) cause disease in horses. A procedure for rapid differentiation of the two EHV-1 subtypes with monoclonal antibodies was developed. Subtype-specific pools of monoclonal antibodies were constructed, characterized, and used in enzyme immunofiltration and indirect immunofluorescence assays to subtype 50 epizootiologically unrelated field isolates of EHV-1. Both assays allowed accurate subtype identification of each EHV-1 isolate with the monoclonal antibody pools. The subtyping procedures were simple and amenable to typing many isolates at one time and permitted unambiguous EHV-1 subtype identification with 3 h after isolation of the virus in tissue culture.

A new field strain of equine abortion virus (equine herpesvirus-1) among Kentucky horses.

From restriction endonuclease characterization of the DNA of 317 isolates of equine abortion virus (equine herpesvirus-1; EHV-1) from 176 epizootically unrelated outbreaks of equine virus abortion occurring over 24 years in Kentucky, an epizootic pattern and variation of the virus have emerged. Two electropherotypes of EHV-1 (1P and 1B) accounted for greater than 90% of the nonvaccine-related abortion isolates examined. From 1960 to 1981, EHV-1 1P was the predominant isolate circulating in the central Kentucky area and the cause of greater than 80% of EHV-1-related abortions. In 1981, the occurrence of isolate 1B-related abortions began to increase and since 1982, 1B has become the most frequently recovered isolate of EHV-1 from aborted fetuses.

Alterations in the equine herpesvirus 1 genome after in vitro and in vivo virus passage.

The effect of in vitro and in vivo serial virus passage on the genetic stability of equine herpesvirus 1 (EHV-1) was investigated by restriction endonuclease analysis of the viral DNA. DNAs of EHV-1 isolates at different passage levels in cultured cells or in Syrian hamsters were compared by electrophoresis of the DNA cleavage fragments produced by restriction endonuclease digestion. No changes were observed in the restriction profile of the DNAs of EHV-1 strains after 100 sequential passages in cultured equine cells. However, serial passage of the virus in hamsters or in cells of non-equine origin quickly gave rise to alterations in the viral DNA. These changes occurring in the restriction endonuclease profiles of EHV-1 DNA during serial virus passage in non-equine cells or animals hosts could be explained by sequence additions or deletions to preexisting restriction fragments resulting in variation in their electrophoretic mobilities.

Application of a chemically inactivated, adjuvanted vaccine to control abortigenic infection of mares by equine herpesvirus I.

A chemically inactivated, adjuvanted vaccine prepared from a virulent strain of Equine herpesvirus I (EHV-I) was used to immunize pregnant Thoroughbred broodmares during a five-year field test designed to determine its safety and efficacy. Each mare in the vaccinated groups received 3 intramuscular injections of vaccine beginning immediately prior to and during the last half of pregnancy. Vaccine was injected at approximately 60-day intervals. The accumulated incidence of EHV-I abortions among vaccinated mares during the field trial period was 1.6/1000 as compared with an incidence of 6.8/1000 in the remainder of the study population.

Serologic and molecular comparisons of several equine herpesvirus type 1 strains.

The molecular and serologic relatedness of 2 recent respiratory tract isolates of equine herpesvirus type 1, designated T1 and T2, were compared with the Army 183, Kentucky-A hamster-adapted (KyA-ha), and L-M cell-adapted (KyA-LM) strains. Electrophoresis in polyacrylamide gels revealed differences in virion structural proteins among 4 purified strains. Seven envelope glycoproteins (molecular weight of 93,000, 65,000, 62,000, 60,000, 36,000, 20,000, and 18,000) corresponding to virion proteins 13, 16, 17, 18, 23, 25, and 26a, respectively, found in both the Army 183 and KyA-ha strains had slightly different molecular weight counterparts in both the T1 and T2 isolates, which had identical structural protein profiles. virion protein 19 (58,000 daltons), a nonglycosylated protein, was present in reduced amounts in the respiratory tract isolates, whereas virion protein 8a (200,000 daltons) was absent. Virion protein 8a, an envelope glycoprotein, was only present in the KyA-ha strain. The T1 and T2 isolates were not neutralized by equine herpesvirus type 2 antiserum and revealed little cross-neutralizatio with the Army 183 and KyA-ha strains in plaque-reduction neutralization tests. Restriction endonuclease cleavage maps of viral DNA revealed a similar, but not identical, number and size of DNA fragments between T1 and T2 isolates. Likewise, DNa profiles of Army 183, KyA-ha, and KyA-LM were also similar to each other, but vastly different from the respiratory tract isolates.

Serologic responses of pregnant thoroughbred mares to vaccination with an inactivated equine herpesvirus 1 vaccine.

The immunogenic potency and safety of a chemically inactivated equine herpesvirus 1 vaccine with added adjuvant was evaluated by testing serum-neutralizing and complement-fixation antibody responses of pregnant Thoroughbred mares. The vaccinated population comprised 321 pregnant mares on 7 farms; 3 in Normandy, France; 1 in Kildare, Ireland; and 3 in central Kentucky. The pattern of antibody response to vaccination was found qualitatively and quantitatively similar to that of pregnant mares previously vaccinated and determined by challenge exposure to be immune to abortigenic infection under experimentally controlled conditions. The safety of the vaccine was demonstrated by the occurrence of only 2 local untoward reactions to the administration of 941 IM injections.

Epidemiological observations on contagious equine metritis in Kentucky, 1978.

Contagious equine metritis, introduced by importation of 2 comtaminated stallions from France, affected 54 Thoroughbred brood mares during the 1978 breeding season in Kentucky. The infection was diagnosed bacteriologically and by the use of a complement fixation test. Although lateral spread to stallions, and probably to a few mares, occurred through human agency in the breeding sheds of 2 stud farms, control measures instituted early in the epidemic confined the disease to brood mares bred by stallion on only these farms.

Replication of equine herpesvirus type 3: kinetics of infectious particle formation and virus nucleic acid synthesis.

The kinetics of equine herpesvirus type 3 (EHV-3) multiplication and of the synthesis of EHV-3 specific DNA and RNA were investigated. A one-step growth curve of EHV-3 in equine epithelial cells from a transitional cell carcinoma was characterized by: (1) a short eclipse period (4 h); (2) an exponential increase in infectious virus between 5 and 10 h post-inoculation; and (3) a slow, inefficient release of newly formed virus into the extracellular fluid. Two hours after infection of cells with EHV-3, the rates of incorporation of specific precursors into total cell RNA or DNA were reduced to 30% and 10%, respectively, of that seen in uninfected cells. With the aid of DNA-RNA hybridization and caesium chloride isopycnic centrifugation techniques, the rates of synthesis of EHV-3 specific nucleic acids at different stages of the virus replication cycle were determined. Virus RNA and DNA synthesis was detectable 2 h after infection and reached maximum levels at an interval (4 to 7 h post-inoculation) corresponding to that period of the virus replication cycle just preceding the time of maximal synthesis of infectious virus.

Immunoglobulins produced by the antigenized equine fetus.

The foal is born without detectable antibody and except for small amounts of IgM is devoid of immunoglobulins. Intrafetal administration of either Venezuelan equine encephalomyelitis virus (VEE-TC83) or ovine erythrocytes elicited IgGa, IgGb and a trace of IgG(T). The fetal blood VEE-TC83 neutralization titre was higher than the neutralization titre elicited by the same preparation in older horses.