OIE Reference Laboratory Reports
Activities in 2011
Name of disease (or topic) for
which you are a designated OIE
Reference Laboratory:
Address of laboratory:
Equine influenza
Irish Equine Centre
Johnstown, Naas, Co. Kildare
IRELAND
Tel.:
(353.45) 86.62.66
Fax:
(353.45) 86.62.73
e-mail address:
acullinane@equine-centre.ie
website:
www.irish-equine-centre.ie
Name (including Title and
Position) of Head of Laboratory
(Responsible Official):
Prof. Ann Cullinane
Name(including Title and
Position) of OIE Reference
Expert:
Prof. Ann Cullinane
Head of Virology
Name (including Title and
Position) of writer of this report
(if different from above):
Annual reports of OIE Reference Centres, 2011
As above
1
Equine influenza
Part I: Summary of general activities related to the disease
1.
2.
Test(s) in use/or available for the specified disease/topic at your laboratory
Test
For
Specificity
Total
HI
Antibody
Influenza subtype
567
SRH
Antibody
Influenza subtype
172
RT-PCR
Virus detection in Nasal Swabs
Influenza type A
130
Embryonated eggs
Virus isolation of RT-PCR positive samples
Influenza
5
Nucleotide Sequencing
Molecular characterisation
Strain
2
Production and distribution of diagnostic reagents
Type of reagent
Antigens for HI tests
Amount supplied nationally
(including for own use)
Amount supplied to other
countries
A/eq/Prague/56 (200mls)
A/eq/Kildare/89 (10mls)
A/eq/Meath/07 (20mls)
5mls to South Africa
1.25mls to South Africa
Antigens for SRH
A/eq/Prague/56 (30mls)
A/eq/Newmarket/2/93 (30mls)
A/eq/Meath/07 (30mls)
A/eq/SouthAfrica/4/03 (40mls)
Control positive serum
15mls
2mls to South Africa
Control negative serum
2mls
1ml to South Africa
Live viruses
1ml to South Africa
A/eq/Meath/07
A/eq/Laois/07
A/eq/Donegal/09
A/eq/Kildare/10
2
1ml to Bulgaria
1ml to Germany
1ml to Bulgaria
1ml to Germany
1ml to Bulgaria
1ml to Germany
Annual reports of OIE Reference Centres, 2011
Equine influenza
Part II: Activities specifically related to the mandate
of OIE Reference Laboratories
3.
International harmonisation and standardisation of methods for diagnostic testing or the
production and testing of vaccines
a)
b)
4.
Establishment and maintenance of a network with other OIE Reference Laboratories
designated for the same pathogen or disease and organisation of regular inter-laboratory
proficiency testing to ensure comparability of results

Interlaboratory comparison of SRH – a panel of equine sera was obtained from Dr Debra Elton of the
OIE reference laboratory at the Animal Health Trust (AHT) in Newmarket. There was good correlation
between the two laboratories.

Agreement was reached with Dr Elton to co-fund the production of strain specific ferret sera at the
AHT. These sera will be used for antigenic characterisation of field isolates.
Organisation of inter-laboratory proficiency testing with laboratories other than OIE
Reference Laboratories for the same pathogens and diseases to ensure equivalence of
results.

Supplied a panel of sera for HI proficiency testing to South Africa.

The compilation of a panel of samples for the second ring trial for the detection of equine influenza by
RT-PCR is in progress. This panel will be distributed both to the original participants in laboratories in
Europe, North America and Australia and to other laboratories in Asia and elsewhere, that have
expressed an interest in participating.
Preparation and supply of international reference standards for diagnostic tests or vaccines
We supply reference virus strains including clade 1 and 2 viruses of the Florida sublineage on request. We also
supply positive and negative equine sera on request. See section 2 above.
5.
Research and development of new procedures for diagnosis and control
Control Procedures
Epidemiological investigations were carried out 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. The results
were published in Equine Veterinary Journal (see section 13 below).
The performance of influenza vaccines in the field was independently evaluated. Many racing authorities, sales
companies and equestrian bodies have mandatory vaccination policies for equine influenza (EI). A field study was
carried out to compare the antibody responses of unvaccinated Thoroughbred weanlings following primary
vaccination in accordance with these regulations, with five different vaccines (two inactivated whole virus, two
subunit and a canary pox recombinant). The antibody response of the horses vaccinated with one of the whole
virus vaccines was significantly higher than that of the horses vaccinated with the other four products. A higher
incidence of poor responders was identified than was reported in previous experimental studies relating to these
products. The results were published in Vaccine (see section 13 below). A second field study demonstrated there
was no significant difference between antibody response induced following booster vaccination of racehorses with
any of six vaccines (three inactivated whole virus, two subunit and a canary pox recombinant). There was a
significant correlation between the antibody level at the time of vaccination and the antibody response. The
findings of the study suggest that equine influenza might be better controlled by monitoring antibody levels and
vaccinating strategically rather than by routine annual vaccination. The results were published in Vaccine (see
section 13 below).
Annual reports of OIE Reference Centres, 2011
3
Equine influenza
6.
7.
Collection, analysis and dissemination of epizootiological data relevant to international disease
control

Chair of the OIE Expert Surveillance Panel (ESP) for equine influenza. The panel consisting of OIE and
WHO experts, met at the OIE headquarters on 24 th of January 2011 to review equine influenza activity in
2010 and make recommendations on the composition of equine influenza vaccines. The meeting was also
attended by observers from laboratories other than OIE equine influenza reference laboratories, from Japan,
Australia, USA and France. A representative from OFFLU was also present. Data relating to viruses isolated
in France, Ireland, the UK and a quarantine facility in Japan were considered. With one exception (a clade 1
virus identified in Ireland) all of the viruses characterised in 2010 belonged to clade 2 of the Florida
sublineage. The recommendations of the ESP were that vaccines for the international market should contain
both clade 1 and clade 2 viruses of the Florida sublineage. The conclusions and recommendations were
published in the OIE Bulletin 2011-2.

Throughout 2011 equine influenza was diagnosed, outbreaks investigated and virus strains characterised by
our laboratory as part of the Equine Influenza Surveillance Programme which is funded by the Department of
Agriculture, Food and the Marine under the National Development Programme. Outbreaks were reported to
the International Collating Centre and all data will be presented to the ESP when the vaccine composition is
reviewed in February 2012.
Maintenance of a system of quality assurance, biosafety and biosecurity relevant to the
pathogen and the disease concerned.
The Irish Equine Centre maintains OECD Good Laboratory Practice (GLP) compliance and the single radial
haemolysis test for equine influenza is one of the many tests accredited to ISO 17025.
8.
Provision of consultant expertise to OIE or to OIE Member Countries


9.
Amended (based on the comments of OIE Members) the chapter on Equine Influenza for the OIE Manual of
Diagnostic Tests and Vaccines for Terrestrial Animals: updated for the seventh edition.
Attended One Flu Strategic Retreat at invitation of CDC, Atlanta. The aim of the meeting was to foster
transdisciplinary collaboration on influenza viruses.
Provision of scientific and technical training to personnel from other OIE Member Countries
An OIE twinning project between the Irish Equine Centre and Harbin Veterinary Research institute, China
commenced in February 2011. The ultimate aim of the project is that Harbin Veterinary Research Institute will
achieve OIE Reference status and implement an effective equine influenza surveillance programme. There is
currently no OIE reference laboratory for equine influenza outside of Europe and North America. In October two
scientists from Harbin Veterinary Research Institute visited the Equine Centre for training in serology and virus
isolation. Techniques included haemagglutination inhibition, single radial haemolysis, virus isolation in eggs and
cell culture, haemagglutination and virus typing. There was also preliminary training in the two quality systems in
place at the Centre, GLP and ISO17025.
In November a veterinary scientist from the Guangzhou Animal Health Inspection Institute, in the Equine Diseasefree Zone in China, visited the laboratory for a two month training programme. Training included
haemagglutination inhibition, single radial haemolysis, virus isolation in eggs and quantification by
haemagglutination, in addition to an introduction to GLP and ISO17025.
10. Provision of diagnostic testing facilities to other OIE Member Countries
Diagnostic testing was provided to Israel, Turkey and France.
4
Annual reports of OIE Reference Centres, 2011
Equine influenza
11. Organisation of international scientific meetings on behalf of OIE or other international bodies
Organised and chaired meeting of the OIE Expert Surveillance Panel (ESP) for equine influenza and the
International Federation for Animal Health (IFAH) to discuss the basis for the recommendations made by the ESP,
the methods employed to assess vaccine efficacy and ways to expedite the updating of the virus strains in the
vaccines. The meeting was held at the OIE on the 9th of December 2011. It was attended by Rod Daniels (WHO,
Mill Hill), Frédéric Descamps (Pfizer), Debra Elton (OIE expert, AHT), Keith Hamilton (OFFLU), Wim
Hesselink (MSD), David John (IFAH), Gounalan Pavade (OIE) and Saskia Van De Zande (MSD). Ely Bénére
(Pfizer), Tom Chambers (OIE expert, Gluck Centre, Kentucky) Jacques Lechenet (Merial) and Jules Minke
(Merial) joined by phone. The meeting was organised with assistance from David John, Keith Hamilton and
Gounalan Pavade. It was proposed that the IFAH would assist the ESP with the evaluation of the potential role of
equine sera in the antigenic characterisation of virus isolates, that the ESP would increase the emphasis on the
investigation of vaccination breakdown in the field and that greater communication between the ESP, the IFAH
and the regulatory authorities might assist with the updating of vaccines in a more timely manner. The minutes of
the meeting are being compiled for circulation by David John (IFAH).
12. Participation in international scientific collaborative studies
Collaborative equine influenza modelling study involving analysis of viral load and interferon response, with the
Department of Mathematics and Statistics, Oakland University, Rochester, Michigan, USA (coordinator Dr Libin
Rong) and the Department of Theoretical Biology and Biophysics, Los Alamos National Laboratory, Los Alamos,
New 11 Mexico, USA. A mathematical model including both innate and adaptive immune responses was
developed to study the within-host dynamics of equine influenza virus infection in horses. The results have
recently been submitted for publication.
13. Publication and dissemination of information relevant to the work of OIE (including list of
scientific publications, internet publishing activities, presentations at international conferences)

Presentations at international conferences and meetings
Presentations at :

Harbin Veterinary Research Institute, China (Ann Cullinane and Sarah Gildea, February 2011)

AVTRW Nottingham UK (Sarah Gildea, April 2011)
Gildea S, Arkins S and Cullinane A (2011). A comparative antibody study of the potential susceptibility of
Thoroughbred and non-Thoroughbred horse populations in Ireland to equine influenza virus. Proceedinngs of
the British Society of Animal Science and the Association of Veterinary Teaching and Research Work. April
2011, Volume 2 Part 1 pp 24. Cambridge University Press.

Palermo Racetrack, Buenos Aires, Argentina. (Ann Cullinane ,October 2011)

BEVA meeting, Lisburn, Northern Ireland. UK. (Ann Cullinane , November 2011)

Scientific publications in peer-reviewed journals

Gildea S, Arkins S, Walsh C, Cullinane A. (2011) A comparison of antibody responses to commercial equine
influenza vaccines following annual booster vaccination of National Hunt Horses-a randomised blind study
Vaccine 29 (22), 3917-39122.

Gildea S, Arkins S, and Cullinane A. (2011) Management and environmental factors involved in equine
influenza outbreaks in Ireland 2007-2010. Equine Veterinary Journal 43 (5), 608-617

Gildea S, Quinlivan M, Arkins S, and Cullinane A. (2011) The molecular epidemiology of equine influenza in
Ireland from 2007-2010 and its international significance. Equine Veterinary Journal (Epub ahead of
print)
Annual reports of OIE Reference Centres, 2011
5
Equine influenza

Gildea S, Arkins S, Walsh C, Cullinane A. (2011) A comparison of antibody responses to commercial equine
influenza vaccines following primary vaccination of Thoroughbred weanlings- A randomised blind study
Vaccine 29 (49), 9214-9223.

Other communications
Recommendations re vaccination of competition horses with updated equine influenza vaccines for discussion and
distribution by Fédération Équestre Internationale (commonly known as the FEI), or in English - the International
Federation for Equestrian Sports.
_______________
6
Annual reports of OIE Reference Centres, 2011