Recent spread of a new strain (A-Iran-05) of foot-and-mouth disease virus type A in the Middle East.

This report describes the characterization of a new genotype of foot-and-mouth disease virus (FMDV) type A responsible for recent FMD outbreaks in the Middle East. Initially identified in samples collected in 2003 from Iran, during 2005 and 2006 this FMDV lineage (proposed to be named A-Iran-05) spread into Saudi Arabia and Jordan and then further west into Turkey reaching European Thrace in January 2007. Most recently A-Iran-05 has been found in Bahrain. To the east of Iran, it has been recognized in Afghanistan (2004-07) and Pakistan (2006-07). Throughout the region, this lineage is now the predominant genotype of FMDV serotype A sampled, and has appeared to have replaced the A-Iran-96 and A-Iran-99 strains which were previously encountered. In August 2007, a new A-Iran-05 sub-lineage (which we have called A-Iran-05(ARD-07)) was identified in Ardahan, Turkey, close to the border with Georgia. This new sub-lineage appeared to predominate in Turkey in 2008, but has, so far, not been identified in any other country. Vaccine matching tests revealed that the A-Iran-05 viruses are antigenically different to A-Iran-96 and more like A(22). These findings emphasize the importance of undertaking continued surveillance in the Middle East and Central Asia in order to detect and monitor the emergence and spread of new FMDV strains.

Evaluation of a monoclonal antibody-based approach for the selection of foot-and-mouth disease (FMD) vaccine strains.

Foot-and-mouth disease (FMD) virus exists as seven serotypes within which are numerous variants necessitating careful selection of vaccine strains. Currently, a serological assay system based on the use of polyclonal vaccine antisera is widely used for this selection. However, inherent variability in the matching antisera used makes the tests poorly reproducible and difficult to interpret. In this study, we have explored the possibility of replacing or supplementing the polyclonal antibody (PAb)-based method with one based on use of monoclonal antibodies (MAb). Panels of MAbs raised against two serotype O vaccine strains were examined for reactivity with 22 field viruses, isolated over a 10-year period between 1991 and 2001. Antigenic site 2 was found to comprise more than one epitope. The sequence variation in capsid protein VP2 harbouring antigenic site 2 was analysed and the amino acid residues at positions 79 and 134 appeared to greatly influence the binding of site 2 MAbs. Prediction of antigenic match based on MAb reactivity did not correlate closely with the results of a PAb-based "gold-standard" method and it was concluded that a wider panel of MAbs are needed that recognise all protective epitopes present on the surface of FMD virus together with a better understanding of those epitopes which are important in conferring protection.

Interferon-gamma production in vitro from whole blood of foot-and-mouth disease virus (FMDV) vaccinated and infected cattle after incubation with inactivated FMDV.

Studies were performed to determine whether a rapid method to detect cell mediated immune responses to foot-and-mouth disease virus (FMDV) could be used either as a diagnostic test or provide a correlate of protection in animals post-vaccination. Using protocols based on the BOVIGAM assay for tuberculosis, whole blood samples from FMDV vaccinated or control animals, before and after challenge infection, were stimulated overnight with inactivated FMDV antigen. The quantity of interferon gamma (IFN-gamma) produced in the supernatants was measured using an ELISA. Specific induction of IFN-gamma was detected in samples from vaccinated, infected and vaccinated-then-infected cattle. Further development of this assay may provide a useful tool for the diagnosis of FMDV immune animals, including the identification of vaccinated animals that have been subsequently infected with FMDV. In these studies, combining the results of the IFN-gamma assay with virus neutralising antibody titre, in groups of vaccinated animals, provided a correlation with the capacity to control virus replication after subsequent challenge.

Evidence that high potency foot-and-mouth disease vaccine inhibits local virus replication and prevents the "carrier" state in sheep.

The ability of a single administration of a high, medium and low potency foot-and-mouth disease (FMD) vaccine to decrease or inhibit local virus replication and excretion in the oropharynx of sheep following aerosol challenge with homologous live virus 14 days later was examined. Unvaccinated sheep showed signs of clinical FMD, whereas all of the vaccinated sheep, regardless of antigen payload, were protected against clinical disease and development of viraemia. Virological and serological results confirmed that there had been no local virus replication in the oropharynx of sheep from the high potency vaccine group in contrast to moderate or substantial virus replication in the oropharynx of the low potency vaccinated or unvaccinated sheep respectively. The vaccines showed no evidence of promoting a local mucosal antibody response at the time of virus challenge, but were capable of stimulating a systemic gamma interferon response, the level of which was related to the antigen payload. This suggests that the systemic gamma interferon response could be a useful indicator of the ability of a FMD vaccine to elicit a sterile immunity and indicates that further work is warranted to investigate the role of systemic gamma interferon in this immunity. This is the first experiment to clearly show that high potency, high payload, FMD vaccines are capable of inhibiting local virus replication and consequently persistence and the carrier state in this target species.

Longevity of antibody and cytokine responses following vaccination with high potency emergency FMD vaccines.

The ability of high potency emergency foot-and-mouth disease (FMD) vaccines to promote sustainable immune responses in sheep and pigs following a single application was examined. All vaccine formulations induced a rapid seroconversion in both species, as expected, which was maintained at near peak titres for up to 6 months in sheep and 7 months in pigs. The Montanide ISA 206 formulation gave the best results in sheep. Vaccinated pigs challenged with homologous FMDV were protected from disease at 7 months post vaccination. Systemic levels of cytokines IL-6, IL-8, and in some pigs IL-12, increased following vaccination and were often maintained at an increased level for the duration of the trials. These initial results suggest that high potency vaccines may promote longer lasting immunity than the conventional lower potency vaccines in ruminants and a comparable response in pigs. Results indicate that in an outbreak situation, should emergency vaccination be done with these high potency vaccines, protection should be conferred for a long enough period for the outbreak to be brought under control without the need to revaccinate. Given the increased interval for re-vaccination the use of high potency vaccines for routine prophylactic campaigns could provide a more cost-effective and efficient means of maintaining herd immunity and is an area thus worthy of further examination.

Stratified and cryogenically stored (SACS) vaccines, a new concept in emergency foot-and-mouth disease vaccine formulation and storage.

Strategic reserves of foot-and-mouth disease (FMD) antigen have become an integral part of FMD control policy for many countries. They are based on two principles, ready formulated vaccine stored at +4 degrees C, or concentrated antigen preparations held at ultra-low temperature for later formulation. However, the latter is more economical, since ready formulated vaccine, based on oil or aluminium hydroxide/saponin adjuvants, requires regular replacement. This is primarily the result of the vaccine's limited shelf-life, nominally 18 months at +4 degrees C. Unfortunately, lowering the temperature of storage, in a bid to extend its shelf-life, has a detrimental effect on the vaccine's potency. Montanide ISA 206 and 25, two 'ready-to-formulate' oil adjuvants which can be used in all target species, are ideal for emergency vaccination. Their potential is enhanced by the ease in which they are formulated into oil emulsion vaccines. Here we describe a novel approach of layering the individual components of FMD vaccine in the same primary container and then storing the product at ultra-low temperature. This avoids the detrimental effect on potency, normally observed with frozen formulated FMD vaccine, and could substantially extend the products shelf-life. The implications of this approach for emergency vaccination strategy are discussed.

The suitability of the 'emergency' foot-and-mouth disease antigens held by the International Vaccine Bank within a global context.

The International Vaccine Bank (IVB) based at the Institute of Animal Health (IAH) in Pirbright, United Kingdom (UK), routinely monitors the suitability of the currently held strains of foot-and-mouth disease (FMD) vaccine virus, in anticipation that vaccine may be required to control FMD outbreaks that pose a threat to member countries. Using primarily the two-dimensional micro-neutralisation test (VNT), bovine polyclonal sera raised against each of the seven current 'emergency' antigens were utilised to measure the relationship of IVB stocks to selected field isolates. The 'O' serotypes, Manisa and Lausanne, exhibited adequate levels of cross-protection against most of the type 'O' field isolates examined. A(22) Iraq 24/64 showed the broadest spectrum of reactivity against the type 'A' field isolates examined and was supplemented by A(15) Thailand 1/60. Some type 'Asia1' field isolates, particularly those from South East Asia, showed antigenic difference to the Asia1 India 8/79 vaccine strain by VNT, but in-vivo testing in the guinea pig model indicated this to be insignificant. The only 'C' serotype representative, C(1) Oberbayern, may be one of the least antigenically diverse of the current portfolio of bank antigens. Comparison of the serological and sequence data shows that despite significant genetic variation between the field isolates examined the antigens held by the IVB should still prove efficacious in the field.