Accelerated vaccination schedule provides protective levels of antibody and complete herd immunity to equine influenza

Reasons for performing study: During the 2007 Australian equine influenza (EI) outbreak, an accelerated primary course 14 day intervaccination schedule was proposed, but not widely implemented. Expert opinion was divided as to the efficacy of such a schedule given the lack of published data. This study determined the level and duration of humoral immunity following administration of a recombinant canarypox‐vectored vaccine (ALVAC‐EIV) with a primary intervaccination interval of 14 days and booster at 105 days. Objectives: To examine whether protective levels of immunity of adequate duration were achieved following a primary course reduced from a minimum interval of 28 to 14 days. Antibody responses to 2 H3N8 American lineage virus strains (including A/equine/Sydney/6085/2007) were assessed and compared to previous challenge studies using ALVAC‐EIV at conventional intervaccination intervals. Methods: Fourteen Thoroughbred horses and 2 ponies from a rural racehorse training property in Victoria, Australia, were vaccinated with ALVAC‐EIV on Days 0, 14 and 105. Serial blood samples were collected over the next 32 weeks and tested with haemagglutination inhibition and single radial haemolysis (SRH) in full assays to evaluate the serological response. Results: All horses and ponies responded to the accelerated ALVAC‐EIV vaccination schedule. Mean SRH antibodies remained above those consistent with clinical protection for the duration of the study period. All vaccinates demonstrated high SRH antibodies 14 days following V2, thereby achieving 100% herd immunity to homologous viral challenge. Conclusions: An accelerated vaccination schedule conferred long‐lasting protective antibody levels despite a >50% reduction in the recommended V1–V2 interval. Potential relevance: High levels of rapidly acquired herd immunity are critical in containing an outbreak of such a highly contagious pathogen as EIV. In a strategic vaccination programme, it is important that horses remain protected for sufficient time to allow control programmes to succeed. An accelerated 14 day primary course intervaccination interval and booster at 105 days achieves both of these objectives.     

Management and environmental factors involved in equine influenza outbreaks in Ireland 2007-2010

Reasons for performing study: Outbreaks of equine influenza (EI) in endemic populations continue to cause economic loss despite widespread vaccination. Hypothesis: To identify the key management and environmental factors that determine the risk of horses contracting EI in an endemic country and to identify control strategies. Methods: Real time‐polymerase chain reaction (RT‐PCR), virus isolation and haemagglutination inhibition were carried out on nasopharyngeal swabs and clotted blood samples collected from horses and ponies showing signs of respiratory disease. On premises where a diagnosis of EI was confirmed, the attending veterinary surgeon was asked to participate in an epidemiological investigation. Results: Between June 2007 and January 2010, EI outbreaks were diagnosed on 28 premises located in 13 of the 32 counties of Ireland. Veterinary advice was sought on average more than 5 days after the first clinical signs were observed. The majority of diagnoses were made by RT‐PCR. Data from 404 horses on 16 premises were used in the epidemiological analysis. In 15 premises, EI was identified following movement of horses. Housing type, teaser stallions or fomites/personnel contributed to virus spread. Vaccination status, number of years vaccination, time since last vaccination and age influenced disease expression. Isolation and vaccination were effective control measures on the premises where they were implemented. Conclusions: Preventative measures include: isolation, clinical monitoring, serological testing and vaccination of new arrivals, booster vaccination of horses at 6 monthly intervals, maintenance of effective boundaries between equine premises and avoidance of stabling in single air spaces. Control measures include: prompt isolation of suspected cases, rapid diagnosis by RT‐PCR, booster vaccination of cohorts and implementation of biosecurity measures to avoid transmission by fomites and personnel. Potential relevance: Implementation of these preventative and control measures should reduce the economic losses associated with outbreaks of EI.     

Poor performance associated with equine gastric ulceration syndrome in four Thoroughbred racehorses

Equine gastric ulceration syndrome (EGUS) is commonly recognised in Thoroughbred racehorses. Although EGUS has previously been associated with reduced athletic performance, no objective studies have been reported. This case report describes 4 racehorses referred for investigation of poor athletic performance where EGUS was the only abnormal finding during a thorough investigation of all body systems. All horses showed considerable improvement in performance following treatment with omeprazole. Therefore, this is the first report in which evidence is presented suggesting a direct link between EGUS and decreased performance, other causes of poor performance having been excluded.     

Myofibroblastic fibrosarcoma with multifocal osseous metaplasia at the site of equine influenza vaccination

We describe a fibrosarcoma in a 12-year-old Quarterhorse × Arabian gelding as a sequela to equine influenza vaccination. Shortly after the second vaccination, swelling at the site was noticed by the owner and it continued to increase in size over the following 6 months. Biopsy of the mass indicated a fibrosarcoma had developed at the vaccination site. It was approximately 20 cm in diameter and elevated well above the level of the skin. There was no clinical evidence of metastases to the lungs or local lymph nodes. Surgical resection of the mass was performed and the wound healed by first and second intention. Histopathological examination and immunohistochemical staining confirmed a myofibroblastic fibrosarcoma with multifocal osseous metaplasia. To the authors' knowledge, this is the first equine case of a vaccine-associated fibrosarcoma.     

Facing the threat of equine influenza

Despite the availability of vaccines, equine influenza virus (EIV) continues to pose a threat to the racing industry. The virus spreads rapidly in unprotected populations and large scale outbreaks, such as those in South Africa in 2003 and Australia in 2007, can cost billions of pounds. Like other influenza viruses, EIV undergoes antigenic variation, enabling it to evade antibodies generated against previous infection or vaccination. The UK has an active surveillance programme to monitor antigenic drift and participates in an international collaboration with other countries in Europe, Japan and the USA to select suitable vaccine strains. Selection is primarily based upon characterisation of the viral haemagglutinin (HA), the surface protein that induces a protective antibody response; this protein is an important component of commercial vaccines. In recent years vaccine technology has improved and diagnostic methods have become increasingly sensitive, both play a crucial part in facilitating the international movement of horses. Mathematical modelling techniques have been applied to study the risk factors involved in outbreaks and provide valuable information about the impact of vaccination. Other factors, such as pathogenicity, are poorly understood for EIV yet may play an important role in the spread of a particular virus. They may also affect the ability of the virus to cross the species barrier, as seen with the transfer to dogs in the USA. Severity of infection is likely to be influenced by more than one gene, but differences in the NS1 protein are believed to influence the cytokine response in the horse and have been manipulated to produce potential vaccine strains.     

Antibody responses after vaccination against equine influenza in the Republic of Korea in 2013

In this study, antibody responses after equine influenza vaccination were investigated among 1,098 horses in Korea using the hemagglutination inhibition (HI) assay. The equine influenza viruses, A/equine/South Africa/4/03 (H3N8) and A/equine/Wildeshausen/1/08 (H3N8), were used as antigens in the HI assay. The mean seropositive rates were 91.7% (geometric mean antibody levels (GMT), 56.8) and 93.6% (GMT, 105.2) for A/equine/South Africa/4/03 and A/equine/Wildeshausen/1/08, respectively. Yearlings and two-year-olds in training exhibited lower positive rates (68.1% (GMT, 14) and 61.7% (GMT, 11.9), respectively, with different antigens) than average. Horses two years old or younger may require more attention in vaccination against equine influenza according to the vaccination regime, because they could be a target of the equine influenza virus.     

湖北株H3N8亚型马流感病毒HA基因的序列测定及其HA蛋白遗传特性分析

2008年从湖北省分离到1株H3N8亚型马流感病毒A/equine/Hubei/6/08。以2002年美国KENTUKY株为模板设计HA基因测序引物,进行RT-PCR,然后测定该分离株的HA基因核苷酸序列。经NCBI上Blast同源性比较发现,与A/equine/Newmarket/5/2003(H3N8)同源性较高为98.7%。HA蛋白遗传进化分析表明该毒株隶属于H3N8亚型马流感病毒中的美洲系福罗里达亚系。该株与OIE现在推荐的疫苗候选株A/equine/Kentuck-y/5/2002(H3N8)HA1蛋白氨基酸序列比对发现有3处氨基酸替换位点;与OIE以往推荐的疫苗候选株A/e-quine/Kentucky/1/1994(H3N8)比对发现有11处氨基酸替换位点。研究结果表明该分离株可作为中国研制马流感疫苗的候选株。     

Detection of clade 2 equine influenza virus in an adult horse recently imported to the USA

A 4‐year‐old Warmblood mare presented to the William R. Pritchard Veterinary Medical Teaching Hospital, School of Veterinary Medicine, University of California at Davis with bilateral mucoid nasal discharge and pyrexia. The mare had recently been imported from Germany, arriving at a quarantine holding facility 72 h prior to presentation. Based on clinical presentation and culture results of tracheal fluid, the mare was diagnosed with bacterial bronchopneumonia secondary to equine influenza. The equine influenza virus (EIV) identified in the imported mare displayed 99.1% nucleotide homology of the HA1 gene to the prototype Florida sublineage clade 2 isolate A/equine/Richmond/1/2007 (H3N8). This case illustrates the risk of introducing a clade 2 EIV in North America.     

Comparison of two modern vaccines and previous influenza infection against challenge with an equine influenza virus from the Australian 2007 outbreak

During 2007, large outbreaks of equine influenza (EI) caused by Florida sublineage Clade 1 viruses affected horse populations in Japan and Australia. The likely protection that would be provided by two modern vaccines commercially available in the European Union (an ISCOM-based and a canarypox-based vaccine) at the time of the outbreaks was determined. Vaccinated ponies were challenged with a representative outbreak isolate (A/eq/Sydney/2888-8/07) and levels of protection were compared. A group of ponies infected 18 months previously with a phylogenetically-related isolate from 2003 (A/eq/South Africa/4/03) was also challenged with the 2007 outbreak virus. After experimental infection with A/eq/Sydney/2888-8/07, unvaccinated control ponies all showed clinical signs of infection together with virus shedding. Protection achieved by both vaccination or long-term immunity induced by previous exposure to equine influenza virus (EIV) was characterised by minor signs of disease and reduced virus shedding when compared with unvaccinated control ponies. The three different methods of virus titration in embryonated hens’ eggs, EIV NP-ELISA and quantitative RT-PCR were used to monitor EIV shedding and results were compared. Though the majority of previously infected ponies had low antibody levels at the time of challenge, they demonstrated good clinical protection and limited virus shedding. In summary, we demonstrate that vaccination with current EIV vaccines would partially protect against infection with A/eq/Sydney/2888-8/07-like strains and would help to limit the spread of disease in our vaccinated horse population.     

Risk factors for equine influenza serum antibody titres in young thoroughbred racehorses given an inactivated vaccine

Young Thoroughbred racehorses (222 yearlings entering training and 246 2-year-old horses already in training) from eight flat-training yards in Newmarket, UK were used to monitor serological responses to vaccination with an inactivated influenza virus vaccine. Blood samples taken prior to and after vaccination were tested by single radial haemolysis (SRH) to determine antibody titres (expressed as area of haemolysis in mm(2)). Prior to vaccination, yearlings had mean antibody titres (64+/-4 mm(2)) that were approximately half of those of 2-year-olds (115+/-3 mm(2)) and 89% of yearlings and 73% of 2-year-olds had SRH titres <140 mm(2). Extrapolation from experimental and field studies suggests that these levels would not protect against homologous influenza virus infection. Both age-groups showed anamnestic responses to vaccination resulting in similar peak mean titres ( approximately 160+/-2mm(2)) with 67% of yearlings and 73% of 2-year-olds achieving levels > or =140 mm(2). A second dose of vaccine administered a month after the first in yearlings did not increase the mean titre but 75% of horses had levels of antibody > or =140 mm(2). The vaccination history in the official passport of yearlings showed that 23% had no record of previous vaccination and were probably fully susceptible to infection. For yearlings entering training, the important predictors from multiple-regression analyses of SRH titres prior to vaccination were "Time since last vaccination," "Total number of previous vaccines" and "Age at first vaccination." In 2-year-olds and following two doses of vaccine in yearlings, there was no significant relationship between these factors and SRH titre.