Parameterization of the durations of phases of foot-and-mouth disease in pigs.

The global interconnectedness of the pig-production industry and the diversity of foot-and-mouth disease (FMD) viruses (FMDVs) currently circulating, makes modeling disease spread and control in FMD-free areas challenging. However, advances in experimental design and transmission studies create opportunities to strengthen our understanding and ability to model FMD transmission. In the current study, we estimated the duration of defined phases of FMDV infection in pigs by using data from a large collection of controlled in vivo experiments. Because the detection of low-levels of viral RNA does not correspond to infectiousness, an experimentally defined minimum threshold of FMDV RNA shedding in oropharyngeal fluids was used to estimate the onset of infectiousness in experiments in which transmission was not evaluated. Animal-level data were used in Accelerated Failure Time models to assess the effect of experimental design factors in the duration of defined phases of FMDV infection: latent, incubation, pre-clinical infectious, clinical infectious, and total infectious periods. The estimated means of the phases were latent: 25 h (95%CI 21, 29), incubation: 70 h (95%CI 64, 76), pre-clinical infectious: 36 h (95%CI 32, 41), clinical infectious: 265 h (95%CI 258, 272) and total infectious: 282 h (95%CI 273, 290). Virus strains and exposure methods had no significant influence on the duration of latency, incubation, or clinical infectious phases. By contrast, the estimated means of the duration of the pre-clinical infectious and total infectious phases were significantly influenced by virus strains, and the duration of the pre-clinical infectious phase was significantly influenced by exposure methods. This study provides disease parameters based on an estimated threshold of the onset of infectiousness and a probability distribution representing the end of infectiousness. Disease parameters that incorporate experimentally-based quantitative proxies to define phases of FMDV infection may improve planning and preparedness for FMD.

A Vaccine Based on the A/ASIA/G-VII Lineage of Foot-and-Mouth Disease Virus Offers Low Levels of Protection against Circulating Viruses from the A/ASIA/Iran-05 lineage.

The recent emergence and circulation of the A/ASIA/G-VII (A/G-VII) lineage of foot-and-mouth disease virus (FMDV) in the Middle East has resulted in the development of homologous vaccines to ensure susceptible animals are sufficiently protected against clinical disease. However, a second serotype A lineage called A/ASIA/Iran-05 (A/IRN/05) continues to circulate in the region and it is therefore imperative to ensure vaccine strains used will protect against both lineages. In addition, for FMDV vaccine banks that usually hold a limited number of strains, it is necessary to include strains with a broad antigenic coverage. To assess the cross protective ability of an A/G-VII emergency vaccine (formulated at 43 (95% CI 8-230) PD50/dose as determined during homologous challenge), we performed a heterologous potency test according to the European Pharmacopoeia design using a field isolate from the A/IRN/05 lineage as the challenge virus. The estimated heterologous potency in this study was 2.0 (95% CI 0.4-6.0) PD50/dose, which is below the minimum potency recommended by the World Organisation for Animal Health (OIE). Furthermore, the cross-reactive antibody titres against the heterologous challenge virus were poor (≤log10 0.9), even in those cattle that had received the full dose of vaccine. The geometric mean r1-value was 0.2 (95% CI 0.03-0.8), similar to the potency ratio of 0.04 (95% CI 0.004-0.3). Vaccination decreased viraemia and virus excretion compared to the unvaccinated controls. Our results indicate that this A/G-VII vaccine does not provide sufficient protection against viruses belonging to the A/IRN/05 lineage and therefore the A/G-VII vaccine strain cannot replace the A/IRN/05 vaccine strain but could be considered an additional strain for use in vaccines and antigen banks.

Assessment of the Risk of Foot and Mouth Disease among Beef Cattle at Slaughter from East African Production Systems.

Endemic foot and mouth disease (FMD) in East African cattle systems is one factor that limits access to export markets. The probability of FMD transmission associated with export from such systems have never been quantified and there is a need for data and analyses to guide strategies for livestock exports from regions where FMD remains endemic. The probability of infection among animals at slaughter is an important contributor to the risk of FMD transmission associated with the final beef product. In this study, we built a stochastic model to estimate the probability that beef cattle reach slaughter while infected with FMD virus for four production systems in two East African countries (Kenya and Uganda). Input values were derived from the primary literature and expert opinion. We found that the risk that FMD-infected animals reach slaughter under current conditions is high in both countries (median annual probability ranging from 0.05 among cattle from Kenyan feedlots to 0.62 from Ugandan semi-intensive systems). Cattle originating from feedlot and ranching systems in Kenya had the lowest overall probabilities of the eight systems evaluated. The final probabilities among cattle from all systems were sensitive to the likelihood of acquiring new infections en route to slaughter and especially the probability and extent of commingling with other cattle. These results give insight into factors that could be leveraged by potential interventions to lower the probability of FMD among beef cattle at slaughter. Such interventions should be evaluated considering the cost, logistics, and tradeoffs of each, ultimately guiding resource investment that is grounded in the values and capacity of each country.

Self-Reporting of Risk Pathways and Parameter Values for Foot-and-Mouth Disease in Slaughter Cattle from Alternative Production Systems by Kenyan and Ugandan Veterinarians.

Countries in which foot-and-mouth disease (FMD) is endemic may face bans on the export of FMD-susceptible livestock and products because of the associated risk for transmission of FMD virus. Risk assessment is an essential tool for demonstrating the fitness of one's goods for the international marketplace and for improving animal health. However, it is difficult to obtain the necessary data for such risk assessments in many countries where FMD is present. This study bridged the gaps of traditional participatory and expert elicitation approaches by partnering with veterinarians from the National Veterinary Services of Kenya (n = 13) and Uganda (n = 10) enrolled in an extended capacity-building program to systematically collect rich, local knowledge in a format appropriate for formal quantitative analysis. Participants mapped risk pathways and quantified variables that determine the risk of infection among cattle at slaughter originating from each of four beef production systems in each country. Findings highlighted that risk processes differ between management systems, that disease and sale are not always independent events, and that events on the risk pathway are influenced by the actions and motivations of value chain actors. The results provide necessary information for evaluating the risk of FMD among cattle pre-harvest in Kenya and Uganda and provide a framework for similar evaluation in other endemic settings.

Sero-epidemiology of foot-and-mouth disease in Darfur area, Western Sudan.

A total of 367 bovine sera positive to antibodies against non-structural proteins (NSPs) of foot-and-mouth disease (FMD) virus were screened for serotype O, A and SAT2 antibodies using the virus neutralization test (VNT). Sera had been collected in 2016 from North (228) and South (139) Darfur States in Western Sudan, where high and low circulation of FMD virus, respectively, prevailed. Tested sera represented the positive-NSPs portion in a random sample of 669 sera collected from both States. According to standard statistical methods, calculations for serial testing (NSPs ELISA and VNT) were applied to estimate prevalence rates of serotype-specific antibodies in the two States. In each State, approximately 20% of NSPs positive sera failed typing. Prevalence's detected were 49% ± 5% (O), 27% ± 5% (A) and 14% ± 4% (SAT2) in North Darfur State and 27% ± 5% (O), 17% ± 4% (A) and 8.0% ± 3% (SAT2) in South Darfur State. In both States, prevalence rates were significantly higher for serotype O, followed by A then SAT2; the same order that was known in most parts of Sudan. Consistently, estimated prevalence's were statistically significantly higher (P < 0.05) in North Darfur than in South Darfur State. Apart from serotype SAT2, detected prevalence rates were lower or similar to those inside the country in previous occasions. Frequency and pattern of distribution of serotype O prevalence were consistent with its suggested pattern of circulation from the Nile valley to other parts in Sudan and significant within the country's circulation. Alternatively, serotype SAT2 prevalence and distribution in Darfur area were suggestive of sporadic occurrence. However, slightly higher prevalence rates of SAT2 antibodies in Darfur than in neighbouring Kordofan areas in 2013 reflected the wide dissemination of SAT2 ( http://www.wrlfmd.org ) in Sudan in early 2014. Risk of FMD in Darfur seemed to be associated with the movement of animals to the North in the wet season as part of the pastoral system, and with movement related to trade into urban centers more than with pastoralism across the Western borders. Generally, the result presented little evidence to suggest presence of FMD primary endemic foci in Darfur area.

Selective isolation of foot-and-mouth disease virus from coinfected samples containing more than one serotype.

The foot-and-mouth disease virus (FMDV) causes a highly infectious disease of all cloven-footed animals. The RNA genome of the virus continuously evolves, leading to the generation of new strains; this necessitates the selection of new vaccine strains to ensure complete protection. Infection with one FMDV serotype does not provide cross-protection against the other FMDV serotypes. Many of the recovered animals may become carriers of the FMDV, but they still remain susceptible to the other serotypes. Coinfection with multiple FMDV serotypes has been reported and studied to understand the virus evolution. Isolation and characterization of all the involved serotypes in the mixed infection case is essential to understand the molecular evolution of the virus. In this study, two cases of coinfection were studied by selective isolation of each of the FMDV serotypes under the cross-serotype-specific immune pressure. It was estimated that the virus present in a minimum of 10-0.92 TCID50 could be isolated from the mixed population containing other serotypes in infective doses of 100.25 TCID50 or less. All involved serotypes present in the mixed infection cases were isolated, without any cross-contamination. Virus characterization revealed that genotype 2 was of serotype A virus from a sample collected in 1995, which was last reported in 1986, indicating a possible subdued prevalence of the genetic group even after vanishing from the field.

Endemic persistence of a highly contagious pathogen: Foot-and-mouth disease in its wildlife host.

Extremely contagious pathogens are a global biosecurity threat because of their high burden of morbidity and mortality, as well as their capacity for fast-moving epidemics that are difficult to quell. Understanding the mechanisms enabling persistence of highly transmissible pathogens in host populations is thus a central problem in disease ecology. Through a combination of experimental and theoretical approaches, we investigated how highly contagious foot-and-mouth disease viruses persist in the African buffalo, which serves as their wildlife reservoir. We found that viral persistence through transmission among acutely infected hosts alone is unlikely. However, the inclusion of occasional transmission from persistently infected carriers reliably rescues the most infectious viral strain from fade-out. Additional mechanisms such as antigenic shift, loss of immunity, or spillover among host populations may be required for persistence of less transmissible strains.

Effective Diagnosis of Foot-And-Mouth Disease Virus (FMDV) Serotypes O and A Based on Optical and Electrochemical Dual-Modal Detection.

Foot-and-mouth disease virus (FMDV) is a highly contagious disease that affects cloven-hoofed animals. The traditional diagnostic methods for FMDV have several drawbacks such as cross-reactivity, low sensitivity, and low selectivity. To overcome these drawbacks, we present an optical and electrochemical dual-modal approach for the specific detection of FMDV serotypes O and A by utilizing a magnetic nanoparticle labeling technique with resorufin ß-d-glucopyranoside (res-ß-glc) and ß-glucosidase (ß-glc), without the use of typical lateral flow assay or polymerase chain reaction. FMDV serotypes O and A were reacted with pan-FMDV antibodies that recognize all seven FMDV serotypes (O, A, C, Asia 1, SAT 1, SAT 2, and SAT 3). The antigen-antibody complex was then immobilized on magnetic nanoparticles and reacted with ß-glc-conjugated FMDV type O or type A antibodies. Subsequently, the addition of res-ß-glc resulted in the release of fluorescent resorufin and glucose owing to catalytic hydrolysis by ß-glc. The detection limit of fluorescent signals using a fluorescence spectrophotometer was estimated to be log(6.7) and log(5.9) copies/mL for FMDV type O and A, respectively, while that of electrochemical signals using a glucometer was estimated to be log(6.9) and log(6.1) copies/mL for FMDV type O and A, respectively. Compared with a commercially available lateral flow assay diagnostic kit for immunochromatographic detection of FMDV type O and A, this dual-modal detection platform offers approximately four-fold greater sensitivity. This highly sensitive and accurate dual-modal detection method can be used for effective disease diagnosis and treatment, and will find application in the early-stage diagnosis of viral diseases and next-generation diagnostic platforms.

Amber codon is genetically unstable in generation of premature termination codon (PTC)-harbouring Foot-and-mouth disease virus (FMDV) via genetic code expansion.

The foot-and-mouth disease virus (FMDV) is the causative agent of FMD, a highly infectious and devastating viral disease of domestic and wild cloven-hoofed animals. FMD affects livestock and animal products' national and international trade, causing severe economic losses and social consequences. Currently, inactivated vaccines play a vital role in FMD control, but they have several limitations. The genetic code expansion technology provides powerful strategies for generating premature termination codon (PTC)-harbouring virus as a live but replication-incompetent viral vaccine. However, this technology has not been explored for the design and development of new FMD vaccines. In this study, we first expanded the genetic code of the FMDV genome via a transgenic cell line containing an orthogonal translation machinery. We demonstrated that the transgenic cells stably integrated the orthogonal pyltRNA/pylRS pair into the genome and enabled efficient, homogeneous incorporation of unnatural amino acids into target proteins in mammalian cells. Next, we constructed 129 single-PTC FMDV mutants and four dual-PTC FMDV mutants after considering the tolerance, location, and potential functions of those mutated sites. Amber stop codons individually substituted the selected amino acid codons in four viral proteins (3D, L, VP1, and VP4) of FMDV. We successfully rescued PTC-FMDV mutants, but the amber codon unexpectedly showed a highly degree of mutation rate during PTC-FMDV packaging and replication. Our findings highlight that the genetic code expansion technology for the generation of PTC-FMD vaccines needs to be further improved and that the genetic stability of amber codons during the packaging and replication of FMDV is a concern.

Identification of novel interactions between host and non-structural protein 2C of foot-and-mouth disease virus.

The 2C protein of foot-and-mouth disease virus (FMDV) is reported to play a critical role in the virus replication complex and modulating the host's immune response. However, the underlying molecular intricacies of subversion of cellular machinery remains poorly understood, thus emphasizing the need to study 2C-host interactions. In this study, we identified the host proteins interacting with the 2C using yeast-two hybrid (Y2H) approach, which is one of the most recognized, high-throughput tools to study protein-protein interactions. The FMDV-2C bait was characterized for auto-activation, toxicity, and expression and was found to be suitable for mating with cDNA library. On preliminary screening a total of 32 interacting host proteins were identified which were reduced to 22 on subsequent confirmation with alternative yeast based assays. Amongst these, NMI/2C interaction has been reported earlier by Wang et al. (2012) and remaining 21 are novel interactions. The Reactome analysis has revealed the role of the identified host proteins in cellular pathways exploited by 2C during FMDV replication. We also confirmed interaction of MARCH7, an E3 ubiquitin ligase with 2C using mammalian two-hybrid system and co-immunoprecipitation. This study leads to the identification of novel 2C interacting host proteins which enhance our understanding of 2C-host interface and may provide checkpoints for development of potential therapeutics against FMDV.

Foot-and-mouth disease virus infection in the domestic dog (Canis lupus familiaris), Iran.

BACKGROUND: Foot-and-mouth disease (FMD) is a highly infectious viral disease, recognised to affect animals in the order Artiodactyla. The disease is rarely fatal in adult animals, however high mortality is associated with neonatal and juvenile infection. CASE PRESENTATION: Five puppies died after being fed lamb carcases, the lambs having died during an outbreak of FMD in Iran. Following a post-mortem examination, cardiac tissue from one of the dead puppies was subjected to virus isolation, antigen ELISA, real-time RT-PCR, sequencing and confocal microscopy to assess the presence and characteristics of any FMD virus. The virological and microscopic examination of the cardiac tissue provided evidence of FMD virus replication in the canine heart. CONCLUSIONS: The data generated in this study demonstrate for the first time that FMD virus can internalise and replicate in dogs and may represent an epidemiologically significant event in FMD transmission, highlighting the dangers of feeding diseased animal carcases to other species. The reporting of this finding may also focus attention on similar disease presentations in dogs in FMD endemic countries allowing a better understanding of the prevalence of such events.

Challenges in foot-and-mouth disease virus strain selection as an input to attain broad vaccine intraserotype cross-protection.

Introduction: Vaccination against foot-and-mouth disease virus is regarded as the most effective way to prevent disease. Selection of appropriate vaccine strains is challenging due to lack of cross-protection between serotypes and incomplete protection between some strains within a serotype. Vaccine effectiveness can be affected by vaccine formulation, vaccination approaches, and also by emerging field variants. Therefore, a precise evaluation of the protective capacity of the selected vaccine virus is essential.Areas covered: This article discusses the limitations of currently in use in vitro methods to assess the protective capacity of vaccine strains. It includes the assessment of well-established South American vaccine strains, O1/Campos and A24/Cruzeiro, against outbreaks/emergencies in the continent, as well as against recent isolates from East and Southeast Asia.Expert opinion: In vitro methods, and particularly r1 values, used to evaluate the protective capacity of vaccine strains are not conclusive and do not cover the variety of field scenarios. At present, an option when facing emergencies could be to use well-established vaccine strains with broad antigenic/immunogenic coverage, including conditions that lead to increased coverage such as vaccine formulations and vaccination schemes.

ID1 inhibits foot-and-mouth disease virus replication via targeting of interferon pathways.

Inhibitor of DNA-binding 1 (ID1) protein has been studied intensively for its functions in tumorigenesis and maintenance of stem cell-like properties, but its roles in virus infection are less understood. In the present study, we have clearly shown that the foot-and-mouth disease virus (FMDV) promotes ID1 degradation via Cdh1-mediated ubiquitination to facilitate its replication. Mechanistic investigations reveal Forkhead Box O1 (FOXO1) as an ID1 partner, which suppresses interferon regulatory factors 3 expression and interferon (IFN) production. Further investigation identified that ID1 suppresses FOXO1 transcription activity through HDAC4-mediated deacetylation, promoting IFN production and antiviral immune response. These studies establish a prominent role for ID1 in suppressing FDMV replication, which may be extended to other viruses.

Evidence of combined effect of amino acid substitutions within G-H and B-C loops of VP1 conferring serological heterogeneity in foot-and-mouth disease virus serotype A.

Foot-and-mouth disease virus (FMDV) serotype A exhibits a higher degree of genetic and antigenic diversity resulting in frequent vaccine failure due to serological mismatch between the vaccine and heterologous strains. Currently, knowledge on the molecular basis of antigenic relationships among the FMDVs is limited; nevertheless, intratype antigenic variation due to mutation(s) is widely considered as the main hurdle to appropriate FMD vaccine development. Here, we studied genetic and antigenic variations of four FMDV serotype A isolates, BAN/GA/Sa-197/2013 (BAN-197), BAN/CH/Sa-304/2016 (BAN-304), BAN/DH/Sa-307/2016 (BAN-307) and BAN/DH/Sa-310/2017 (BAN-310) circulating in Bangladesh during 2013-2017. Initially, antigenic relationships (r1 -values) of the field isolates were evaluated by the two-dimensional microneutralization test (2D-MNT) using the hyperimmune antisera raised in cattle against the vaccine strain, BAN-304. Interesingly, the results showed protective serological cross-reactivity (r1 -values > 0.4) between the vaccine strain and the field isolates, BAN-307 and BAN-310, except BAN-197 that substantially mismatched (r1  = 0.129 ± 0.043) with the BAN-304. Although VP1-based phylogeny grouped all the isolates within the same sublineage C (a subgroup of VP3Δ59 variant) under the lineage A/ASIA/G-VII, strikingly, computational analyses of the viral capsid proteins demonstrated significant deviation at the VP1 G-H loop of BAN-197 from the vaccine strain, while VP(2-4) of both isolates were structurally conserved. To bridge the gap of how the distortion of the G-H loop and consequent antigenic hetergeneity occurred in BAN-197, we performed in silico combinatorial substitutions of the VP1 mutant amino acids (aa) of BAN-197 with the respective residues in BAN-304. Remarkably, our analyses revealed that two substitutions of distantly located aa at B-C (T48I:threonine â†’ isoleucine) and G-H (A143V:alanine â†’ valine) loops, in combination, distorted the VP1 G-H loop. Overall, this work contributes to understanding the molecular basis of antigenic relationships operating in serotype A FMDVs and the selection of suitable vaccine strain(s) for effective prophylaxis of FMD based on VP1-based analyses.

Detection of an emerging novel sublineage Ind2001BD1 and lineage PanAsia of foot-and-mouth disease virus serotype O in cattle in Manikgonj district of Bangladesh, 2018.

Background: Foot-and-mouth disease (FMD) is an endemic disease of cloven-hoofed animals in Bangladesh and multiple outbreaks occur every year because of the FMD virus (FMDV). Aim: The aim of the present investigation was to determine the molecular characterization of the VP1 coding region of FMDV serotype O outbreak in cattle. Methods: A total of four tongue epithelial specimens were collected from clinically FMD-positive cattle during June 2018 in Manikgonj district of Bangladesh. Results: All four isolates were recorded positive for FMDV serotype O. The phylogenetic analysis showed that two isolates were clustered within an emerging novel sublineage Ind2001BD1 under lineage Ind2001 of FMDV serotype O, which was identified during 2012-2016 in Bangladesh. One isolate was clustered within the lineage PanAsia of FMDV serotype O and was closely related to an isolate identified in Nepal in 2009. The phylogenetic reconstruction revealed that all the four isolates belong to the Middle East-South Asia topotype. Conclusion: Therefore, multiple lineages of the FMDV serotype O are circulating among the cattle in the outbreak area, which make it more complex for the FMD control program in Bangladesh. A comprehensive study on the genetic characteristics of FMDV across the country is required for effective FMD prevention and control strategy.

A Virus-Like Particle-Based Solid-Phase Competition Enzyme-Linked Immunosorbent Assay for Antibody Detection of Foot-and-Mouth Disease Virus Serotype O.

Foot-and-mouth disease (FMD) is caused by FMD virus (FMDV) is a highly contagious disease of ruminants, which is primarily controlled by vaccination. The monitoring of antisera after vaccination is currently depending on liquid-phase blocking ELISA (LPBE). Recently, bacterium-original FMD virus-like particle (VLP) showed the potential as vaccine candidates. In this study, to minimize the risk of live virus involvement, the Escherichia coli original VLP of FMDV serotype O were used as the immunogen for monoclonal antibodies (Mabs) production and the capture antigen in the development of a solid-phase competition ELISA (SPCE). The samples with a percentage inhibition of >50% were considered positive in the SPCE assay. The concordance rate of the Mab-based SPCE compared with the LPBE for clinical serum samples test was 93.4%, and with a high agreement (kappa = 0.892) with LPBE in antibody duration monitoring. Results indicated that the VLP-based SPCE had high specificity and sensitivity, which provides an alternative method for postimmunization antibody evaluation of FMDV serotype O.

Emergence of foot-and-mouth disease virus serotype Asia1 group IX in India.

Foot-and-mouth disease virus (FMDV) serotype Asia1 is prevalent in India and is responsible for a minor proportion of FMD outbreaks. Globally, serotype Asia1 is grouped into nine different groups (GI-IX) based on genetic analysis. In India, only Asia1/G-III and Asia1/G-VIII have been documented so far. Phylogenetic analysis of recent serotype Asia1 isolates from India revealed the emergence of Asia1/G-IX. The Asia1/G-IX lineage shares recent common ancestry with Asia1/G-VIII dating to 2016. The root state posterior probabilities of Asia1/G-VIII are inclusive and there may have been either an incursion of the virus from Bangladesh, where it was first identified, or in situ evolution of the virus within India, which is an intriguing possibility.

Transmission network reconstruction for foot-and-mouth disease outbreaks incorporating farm-level covariates.

Transmission network modelling to infer 'who infected whom' in infectious disease outbreaks is a highly active area of research. Outbreaks of foot-and-mouth disease have been a key focus of transmission network models that integrate genomic and epidemiological data. The aim of this study was to extend Lau's systematic Bayesian inference framework to incorporate additional parameters representing predominant species and numbers of animals held on a farm. Lau's Bayesian Markov chain Monte Carlo algorithm was reformulated, verified and pseudo-validated on 100 simulated outbreaks populated with demographic data Japan and Australia. The modified model was then implemented on genomic and epidemiological data from the 2010 outbreak of foot-and-mouth disease in Japan, and outputs compared to those from the SCOTTI model implemented in BEAST2. The modified model achieved improvements in overall accuracy when tested on the simulated outbreaks. When implemented on the actual outbreak data from Japan, infected farms that held predominantly pigs were estimated to have five times the transmissibility of infected cattle farms and be 49% less susceptible. The farm-level incubation period was 1 day shorter than the latent period, the timing of the seeding of the outbreak in Japan was inferred, as were key linkages between clusters and features of farms involved in widespread dissemination of this outbreak. To improve accessibility the modified model has been implemented as the R package 'BORIS' for use in future outbreaks.

Shifting to foot-and-mouth disease-free status without vaccination: Application of the PROMETHEE method to assist in the development of a foot-and-mouth national program in Uruguay.

Foot-and-mouth disease (FMD) is highly contagious, and the introduction of FMD virus in countries free of the disease can result in large epidemics, similar to those observed in the United Kingdom, Japan, and Uruguay. Many countries or regions of South America are recognized as "FMD-free with vaccination" or "FMD-free without vaccination." Uruguay has been certified as FMD-free with vaccination, and the transition to the status of FMD-free without vaccination has been discussed among the stakeholders of the Ministry of Livestock Agriculture and Fisheries (MGAP in Spanish). This study illustrates how the Preference Ranking Organization Method for Enrichment Evaluation (PROMETHEE) can identify and rank alternative actions to be used in the national FMD program in Uruguay. It uses multiple-criteria decision analysis (MCDA) to structure the problem comprehensively. This helps to identify critical issues and potential solutions when planning a national FMD program for a country with a disease-free without vaccination status. The analysis highlighted the following perspectives: i) the importance of communication strategies as the country changes to "FMD-free without vaccination" status, as well as potential challenges such as a lack of trust in government; ii) the importance of enhancing epidemiological capacity and data management to allocate resources better and provide feedback from the surveillance system; iii) the need to re-evaluate the animal health compensation strategy to stimulate biosecurity and disease reporting; and iv) the importance of assisting the backyard pig sector given the risks it carries and the complexity of dealing with because of its informality, the educational status of its owners, and the lack of formal assistance by veterinary services. The results suggests that the MCDA approach could be a useful tool for animal health agencies when planning a program to eliminate FMD (as well as other diseases) in a structured way because it helps to identify which alternatives would yield more effective results and it helps to address future challenges.