Isolation of Salem virus, a novel equine paramyxovirus, and assessment of its etiologic role in a disease outbreak
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Etiology
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In this study, we evaluated whether five rapid antigen detection kits for human influenza could be used for the diagnosis of equine influenza (EI). Limiting dilution analyses showed that Directigen Flu A+B and ESPLINE INFLUENZA A&B-N had the highest sensitivities to equine-2 influenza viruses (EIVs) among the kits investigated. From the results of virus detection in nasal swabs taken from horses infected with EIV, these two kits could produce positive results in reasonable agreement with those obtained by virus isolation or RT-PCR, suggesting that these kits could be useful for rapid diagnosis of EI in the field. However, from the viewpoint of specificity for EIV, Espline seems to be superior to Directigen.
Equine influenza
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Simulation models were developed to quantify the likelihood of equine influenza virus infection entering pre-movement isolation, persisting through pre- and post-movement isolation periods without being detected by scheduled laboratory testing, and escaping to infect susceptible horses at a destination. The mean probability of escape ranged from 1 in 1,200,000 to 1 in 600,000 depending on lot size. For 95% of iterations the probability of escape was less than 1 in 200,000, regardless of lot size. For a large group of 600 horses processed as multiple separate lots, the mean probability of escape ranged from 1 in 10,000 to 1 in 56,000 depending on lot size. As a result of this analysis, a modified protocol, with two tests during pre-movement isolation and an additional test during post-movement isolation at the Chief Veterinary Officer's discretion, was implemented.
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We developed the TaqMan based real-time EZ RT-PCR assay for the detection of equine-2 influenza viruses (H3N8). Specificity was tested for various wild strains of equine influenza viruses. Our TaqMan EZ RT-PCR could detect all recent European and American strains isolated after 1989, and historical strains except a variant A/equine/Tokyo/71 strain isolated in 1971. Sensitivity was evaluated to detect 10 EID50 viral RNA using the RNA extracted from hen’s egg allantoic fluid infected with A/equine/La Plata/93 (LP93) strain. Detection sensitivity of TaqMan EZ RT-PCR method from the 180 nasal swab samples collected from horses experimentally infected with LP93 strain was compared with the RT-PCR and egg isolation methods. The results suggested that the TaqMan EZ RT-PCR would be the more sensitive method to detect the equine-2 influenza virus than the egg isolation method and as sensitive as RT-PCR method.
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An equine influenza virus was isolated from horses showing typical influenza symptoms in Fuyun County of Xinjiang province in October 2007. Electron microscope observation and sequence analysis confirmed that the virus belonged to H3N8 subtype and was highly homologous with equine influenza virus American isolates.
Equine influenza
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Summary Processing of nasal materials from clinical cases during the 1987 influenza epidemic in Northern and Central India resulted in the isolation of two haemagglutinating agents; one each from donkeys and horses at Bhiwani in Haryana State and Ludhiana in Punjab State, respectively. These were typed as Influenza A/Equi‐2 viruses by haemagglutination inhibition test. The two isolates were designated as A/Equi‐2/Bhiwani/1/87 and A/Equi‐2/Ludhiana/1/87. The Bhiwani/87 isolate was confirmed to have H3N8 antigenic structure and was indistinguishable from the Miami/63 strain of A/Equi‐2 virus. However, the A/Equi‐2 Ludhiana/87 isolate was closely related to the Fontainebleau/79 strain of A/equi‐2 virus.
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This study aimed to determine whether TNF-α is transferred to equine neonates via colostrum and the relationship between TNF-α and IgG concentrations in the equine neonate. Colostrum, presuckle and postsuckle foal serum samples were collected from healthy mares and their foals. Equine TNF-α ELISA and IgG SRID kits were used to determine the concentrations of TNF-α and IgG, respectively. Statistical analysis was performed using the Spearman rank correlation. TNF-α concentrations in all presuckle foal serum were below the limit of detection in 15/16 foals and increased in postsuckle foal serum to a mean concentration of 7.7 x 10(4) pg/ml. TNF-α concentrations in postsuckle foal serum and colostrum showed significant correlation (rho=0.668; P=0.005). However, TNF-α and IgG concentrations in colostrum or postsuckle foal serum did not correlate (rho<-0.016; P>0.05). Ratios of TNF-α/IgG in colostrum or postsuckle foal serum showed significant correlation (rho=0.750; P=0.0008). These results indicate that TNF-α is transferred to the foal via colostrum absorption and may play a role in early immunity.
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