Subdistrict-Level Reproductive Number for Foot and Mouth Disease in Cattle in Northern Thailand

Foot and mouth disease (FMD) is an important contagious transboundary disease that causes a significant economic loss for several countries. The FMD virus (FMDV) can spread very rapidly by direct and indirect transmission among susceptible animals. The complexity and magnitude of FMDV transmission at the initial stages of the epidemic can be expressed by the basic reproductive number (R0), and furthermore, control strategies can be assessed by the estimation of the effective reproductive number. In this study, we aimed to describe FMD outbreaks among smallholder cattle farms by subdistricts in the northern Thailand and compute the effective reproductive number for outbreaks caused by FMDV serotype O and overall serotypes, including serotype O, serotype A, and unidentified serotype, at the subdistrict level (Rsd) using an epidemic doubling time method. Field data of FMD outbreaks during 2015–2017 that affected 94 subdistricts in northern Thailand were assessed to estimate the Rsd. Results showed that 63.38% (90/142) of the FMD outbreak episodes in cattle were caused by FMDV serotype O. The average doubling time and the Rsd estimated of the outbreaks caused by FMDV serotype O and overall serotype were 2.80 and 4.67 months, and 1.06 and 1.04, respectively. Our results indicated that transmission of FMD in cattle at the subdistrict level in northern Thailand was not controlled (Rsd > 1), which indicates the endemicity of the disease in the region. Although control measures are in place, the results from this study highlighted the need for enhancing FMD monitoring and control strategies in northern Thailand.

A heat-induced Mutation on VP1 of FMDV Serotype O Enhanced Capsid Stability and the Immunogenicity

Foot-and-mouth disease (FMD) is a highly contagious viral disease affecting cloven-hoofed animals that cause a significant economic burden globally. Vaccination is the most effective FMD control strategy. However, FMDV particles are prone to dissociate when appropriate physical or chemical conditions are unavailable, such as an incomplete cold chain. Such degraded vaccines result in compromised herd vaccination. Therefore, thermostable FMD particles are needed for use in vaccines. This study generated thermostable FMDV mutants (M3 and M10) by serial passages at high temperature, subsequent amplification, and purification. Both mutants contained an alanine to threonine mutation at position 13 in VP1 (A1013T), although M3 contained 3 additional mutations. The selected mutants showed improved stability and immunogenicity in neutralizing antibody titers, compared with the wild-type (wt) virus. The sequencing analysis and cryo-electron microscopy showed that the mutation of alanine to threonine at the 13th amino acid (aa) in VP1 protein (A1013T) is critical for the capsid stability of FMDV. Virus-like particles (VLPA1013T) containing A1013T also showed significantly improved stability to heat treatment. This study demonstrated that Thr at the 13th amino acid of VP1 could stabilize the capsid of FMDV. Our findings will facilitate the development of a stable vaccine against FMDV serotype O. IMPORTANCE Foot-and-mouth disease (FMD) serotype O is one of the global epidemic serotypes, and cause significant economic loss. Vaccination plays a key role in the prevention and control of FMD. However, the success of vaccination mainly depends on the quality of the vaccine. Here, the thermostable FMDV mutants (M3 and M10) were selected through thermal-screening at high temperatures with improved stability and immunogenicity compared to the wild-type virus. The results of multi-sequence alignment and Cryo-EM analysis showed that the "Thr" substitution at the 13th amino acid in VP1 protein is critical for the capsid stability of FMDV. For thermolabile type O FMDV, this major discovery will aid the development of its thermostable vaccine.

Development of an indirect chemiluminescence immunoassay using multi-epitope recombinant protein to specifically detect antibodies against foot-and-mouth disease virus serotype O in swine

Foot-and-mouth disease virus (FMDV) has led to serious losses in animal husbandry worldwide. Seromonitoring of FMDV post-vaccination is important for the control and eradication of foot-and-mouth disease (FMD) in vaccinated regions and countries. However, many commercial kits present high false-positive rates. In this study, a multi-epitope-based indirect chemiluminescence immunoassay (ME-CLIA) was developed for specifically detecting antibodies against FMDV serotype O in swine sera. The developed method presented high diagnostic sensitivity, excellent diagnostic specificity, and could detect a broad spectrum of antibodies against FMDV serotype O. The diagnostic performance, accuracy rate, and analytical sensitivity of ME-CLIA were compared with those of three commercial kits. The immune protection value of multiple-epitope recombinant vaccine detected using ME-CLIA was preliminarily determined by observation of clinical symptoms post-immunization challenge, the results of which indicated that the ME-CLIA can be employed as a matching detection method for evaluating multiple-epitope recombinant vaccine. The percent positive values of ME-CLIA determined using swine vaccinated with inactivated vaccine were significantly positively correlated with the titers of liquid-phase blocking ELISA (r = 0.8361, p < 0.0001). These results indicated that ME-CLIA is suitable for detection of antibodies against FMDV serotype O in swine and for potency evaluation of multiple-epitope and inactivated vaccines.

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. Keywords: Cattle, Foot-and-mouth disease, Ind2001BD1, Lineage, PanAsia.

Genome Sequences of Four Foot-and-Mouth Disease Virus SAT 1 Topotype X Isolates from Cameroon

We report the genomes of four foot-and-mouth disease virus (FMDV) serotype SAT 1 topotype X isolates from Cameroon. The viruses were isolated from bovine epithelium collected during an outbreak in 2016. These novel sequences update knowledge of FMDV diversity in Central Africa and contribute to regional FMDV molecular epidemiology.

Implication of Broadly Neutralizing Bovine Monoclonal Antibodies in the Development of an Enzyme-Linked Immunosorbent Assay for Detecting Neutralizing Antibodies against Foot-and-Mouth Disease Virus Serotype O

ABSTRACT Vaccination with inactivated vaccines is still the main measure to control foot-and-mouth disease (FMD) in areas where the disease is endemic, and the level of neutralizing antibody in vaccinated animals is directly related to their protection against virus challenge. Currently, neutralizing antibody is mainly detected using the virus neutralization test (VNT) based on cell culture, which is laborious and time-consuming and requires restrictive biocontainment facilities. In this study, two broadly neutralizing antibodies (bnAbs), E46 and F128, were successfully produced using techniques for the isolation of single B cells from peripheral blood mononuclear cells (PBMCs) from bovines sequentially immunized with three topotypes of foot-and-mouth disease virus (FMDV) serotype O. Based on these bnAbs, a blocking enzyme-linked immunosorbent assay (ELISA) for detecting neutralizing antibodies (NA-ELISA) against FMDV serotype O was developed. The specificity and sensitivity of the test were estimated to be 99.21% and 100%, respectively. A significant correlation (P < 0.01) was observed between the NA-ELISA titers and the VNT titers for all sera from vaccinated animals and for all tested strains, suggesting that the NA-ELISA could detect neutralizing antibodies against FMDV serotype O strains of wide antigenic and molecular diversity and could be used for the evaluation of protective immunity.

First Complete Genome Sequence of the Novel Lineage G-IX (BD-18) of FMDV Serotype Asia1

ABSTRACTFoot-and-Mouth Disease (FMD) has been a major threat to livestock worldwide, and is caused by FMD Virus (FMDV) existing as seven serotypes (A, O, C, Asia1 and SAT 1-3), each having variability into topotypes, genetic lineages, sublineages, and strains. Three serotypes are circulating in Indian subcontinent with the widespread distribution of serotype O, whereas serotype A and Asia1 are restricted to certain geographical regions. During 2017-2018, the Sindh-08 lineages of Asia1 serotype was reported from Pakistan, however, in 2018, a novel circulatory foot-and-mouth disease virus serotype Asia1 BD-18 (G-IX) lineage containing a unique mutation has emerged in Bangladesh. The first complete genome of the Asia1/ASIA/G-IX novel lineage strain, isolated from Bangladesh, is reported here. Amino acid substitutions at critical antigenic sites of capsid were identified compared to genome of existing vaccine strain (IND/63/72), and contemporary FMDV serotype A isolate of Bangladesh.