Mouth Disease Virus Serotype Research Articles

Comparative assessment of β-propiolactone, binary ethyleneimine and formaldehyde in inactivating Foot and Mouth Disease virus serotype O

Comparative assessment of β-propiolactone, binary ethyleneimine and formaldehyde in inactivating Foot and Mouth Disease virus serotype O

A reverse transcription-multiplex PCR strategy devised for concomitant detection and differentiation of foot and mouth disease virus serotypes O, A and Asia 1 in India

Serotype identification occupies the central part of foot and mouth disease (FMD) diagnosis workflow and vaccination decision tree. In this study, a reverse transcription-multiplex PCR (RT-mPCR) strategy wherein three assays with unique combinations of serotype specific primers targeting the VP1 region was developed to differentiate FMD virus serotypes O, A and Asia 1 based on differential size of the PCR amplicons on agarose gel. Their diagnostic performance relative to the mPCR assay in use in India was evaluated on 169 clinical samples and 210 cell culture grown virus isolates. The relative diagnostic sensitivity was found to be 99.69%, 98.78% and 99.08% for primer combinations 1, 2 and 3, respectively. These assays proved their worth by detecting serotype in three FMD suspected specimens that went undiagnosed in the existing mPCR and also by identifying multiple serotypes in the same sample. Their detection limits varied from log10 2 to log10 4 viral RNA dilution and from 100 to 0.1 TCID50 virus depending on the serotype. The validated novel mPCR assays show promise to be included in the routine diagnostic tool-box to augment the efficiency of diagnosis of FMD virus serotypes that display extreme genetic diversity and a tendency of transboundary dispersal.

Emergence of a novel genetic lineage ‘A/ASIA/G-18/2019′ of foot and mouth disease virus serotype A in India: A challenge to reckon with

Foot and mouth disease (FMD) has engendered large scale socioeconomic crises on numerous occasions owing to its extreme contagiousness, transboundary nature, complicated epidemiology, negative impact on productivity, trade embargo, and need for intensive surveillance and expensive control measures. Emerging FMD virus variants have been predicted to have originated and spread from endemic Pool 2, native to South Asia, to other parts of the globe. In this study, 26 Indian serotype A isolates sampled between the year 2015 and 2022 were sequenced for the VP1 region. BLAST and maximum likelihood phylogeny suggest emergence of a novel genetic group within genotype 18, named here as ‘A/ASIA/G-18/2019′ lineage, that is restricted so far only to India and its eastern neighbour, Bangladesh. The lineage subsequent to its first appearance in 2019 seems to have displaced all other prevalent strains, in support of the phenomenon of ‘genotype/lineage turnover’. It has diversified into two distinct sub-clusters, reflecting a phase of active evolution. The rate of evolution of the VP1 region for the Indian serotype A dataset was estimated to be 6.747 × 10−3 substitutions/site/year. India is implementing a vaccination centric FMD control programme. The novel lineage showed good antigenic match with the proposed vaccine candidate A IND 27/2011 when tested in virus neutralization test, while the existing vaccine strain A IND 40/2000 showed homology with only 31% of the isolates. Therefore, in order to combat this challenge of antigenic divergence, A IND 27/2011 could be the preferred strain in the Indian vaccine formulations.

Molecular Characterization of Foot and Mouth Disease Virus Serotype A from Mizoram

Background: Foot and mouth disease virus serotype A has been reported from different states of India and is considered to be the third most commonly encountered serotype besides serotype O and Asia1. Serotype A viruses are most diverse in nature and have been globally grouped into 26 different types. Methods: Foot and mouth disease virus serotype A could be detected from an outbreak in Mizoram that occurred in April 2020. The virus could be confirmed by performing a polymerase chain reaction targeting the 5' UTR region and subsequent serotype-specific multiplex PCR. The full-length VP1 capsid protein gene of the virus could be amplified, cloned and sequenced. Result: Sequence analysis revealed the virus belonging to genotype VII of serotype A. Deduce amino acid sequence comparison it has been observed that the 144R-G-D146tripeptidein the G-H loop remains conserved. However, the present sequence (OL419371), when compared with the Indian vaccine strain of serotype-A(HM854025) as well as the 2001 Mizoram outbreak (AF390659) revealed 11 amino acid differences with the Indian vaccine strain and almost 36 amino acid differences with Mizoram 2001 isolate in different amino acid positions. This indicates the diverse nature of the virus within a geographical area.Background: Foot and mouth disease virus serotype A has been reported from different states of India and is considered to be the third most commonly encountered serotype besides serotype O and Asia1. Serotype A viruses are most diverse in nature and have been globally grouped into 26 different types.

Characterisation of capsid polypeptide P1 and capsid protein VP1 of the Malaysia foot and mouth disease virus (FMDV) serotype O and A isolates

Foot and mouth disease virus (FMDV) is the cause of foot and mouth disease (FMD) outbreaks in livestock worldwide, which affects domestic and international trade, resulting in significant economic losses and social consequences. For efficient monitoring and prevention of FMD outbreaks, the need for improved strategies to control FMDV and achieve FMD-free status with various control measures including vaccination can be established. In vaccinology, major advances and discoveries in vaccination variations including DNA and protein subunit vaccines proved to be more economical and sustainable. To develop a safe vaccine for animals, possible antigenic genes or antigens need to be identified and characterised. The FMDV is a single-stranded RNA virus consisting of a capsid precursor polypeptide, P1, which encodes for four structural proteins (VP4-1), leading to antigenic variation and VP1 potentially carrying the key epitope for vaccine development. This study aims to identify and characterise the capsid polypeptide, P1 and capsid protein, VP1 of the Malaysian FMDV serotype O and serotype A isolates. The nucleotide and protein sequences were identified based on the FMD outbreaks in Malaysia and the antigenicity of the P1 and VP1 was predicted by Kolaskar and Tongaonkar's semi-empirical method. Subsequently, the P1 and VP1 genes were inserted into pET-28a, respectively, and used for protein expression analysis. The P1 and VP1 were predicted to be antigenic via in silico analysis and successfully expressed and characterised through in vitro analysis. Hence, this study can be exploited as a tool to design a new novel vaccine for vaccine development against FMD in bovines.

Optimization of Size Exclusion Chromatography for the Purification and Quantification of Foot and Mouth Disease Virus Serotype “O”

Optimization of Size Exclusion Chromatography for the Purification and Quantification of Foot and Mouth Disease Virus Serotype “O”

EPIDEMIOLOGICAL STATISTICS OF FMD VIRUS SEROTYPES A, O AND ASIA-1 IN PUNJAB, PAKISTAN DURING 2014-2019

Foot and Mouth Disease is an endemic disease and three serotypes (A, O, Asia-1) are prevalent in Pakistan. The present study shows the frequency of Foot and Mouth Disease Virus (FMDV) serotypes (A, O, Asia-1) in Punjab Province of Pakistan by using antigenic Enzyme-Linked Immunosorbent Assay (ELISA). This study was conducted from 2014-2019 by using a total of 184 epithelial samples from field. Out of 184 samples, 106 samples showed positive results. Out of three serotypes, serotype „A‟ was the most prevalent (39.62%) followed by serotype „O‟ (33.96%) while the least prevalent was serotype Asia-1 (26.41%). It was marked that no other serotype of FMD reported during this study. This study also described the year wise prevalence of three prevailing FMD serotypes (A, O, Asia-1). During year 2014, the serotype „A‟ was more prevalent (95.83%) followed by serotype „Asia-1‟ (4.16%) while no case of serotype „O‟ was found. During year 2015, the prevalence trend was almost the same as in 2014. In this year, serotype „A‟ was 55.55% while serotype „Asia-1‟ 37.03% followed by the least prevalent serotype „O‟ as 7.40%. During year 2016, 19 positive cases were reported from which the maximum prevalence was serotype „O‟ and „Asia-1‟ (42.10%) followed by serotype „A‟ (15.78%) as least prevalent. During year 2017, the positive cases for FMD increased remarkably i.e. 29 in number. This year serotype „O‟ was highest in number with prevalence 75.86%, and serotype „Asia-1‟ 20.68% whereas only one case reported for serotype „A‟ with only 3.44% prevalence. During year 2018 and 2019, only 7 cases described with no evident case ofserotype „A‟. During these two years 4 cases of serotype „O‟ were observed followed by 3 cases of serotype „Asia-1‟.

Isolation and molecular characterization of Foot and Mouth Disease virus serotype O circulated in Kenya during the period 2013-2018

The endemicity of Foot-and-mouth disease (FMD) in Kenya has been recognized for over a century, with the first recorded cases dating back to 1915. Production of effective vaccines against incursions of infection in endemic areas is achieved by evaluating the genetic and antigenic characteristics of the circulating viruses. The present study aimed to isolate, serotype, and molecularly characterize FMDV from Kenya from 2013-2018. Isolation was done from 58 field samples on BHK-21 cells, and serotyping of the isolated viruses was carried out using antigen ELISA. Isolated viruses were also analyzed using reverse transcription PCR, and the PCR products were subjected to sequencing. Based on the quality of obtained sequence spectra, only 51 isolates were aligned using MEGA v11.0.8, employing the ClustalW algorithm. SeaView version 5.0.4 was used to edit the alignment, and MEGA 11.0.8 was used to construct the phylogenetic tree and align it with the commercially used vaccinal strains (OK77/78 and OK82/98). With a few exceptions, isolates collected over the same period and those from the same regions consistently clustered in the same lineage or closer to each other. A total of 50/51 strains belong to the East African-2 (EA-2) topotype together with the vaccine strain OK82/98. However, only one strain (1/51) isolated from Tana River county belongs to the EA-1 topotype together with the current vaccine strain (OK77/78). None of these isolates was found to belong to the EA-and EA-4 topotypes. This study emphasizes the importance of regular surveillance and characterization of circulating virus strains for developing effective vaccines against FMD. It’s proposed that future vaccine candidate strains selection could consider EA-2 topotype strains to control FMDV circulating in Kenya.

Evolutionary Dynamics of Foot and Mouth Disease Virus Serotype A and Its Endemic Sub-Lineage A/ASIA/Iran-05/SIS-13 in Pakistan.

Foot and mouth disease (FMD) causes severe economic losses to the livestock industry of endemic countries, including Pakistan. Pakistan is part of the endemic pool 3 for foot and mouth disease viruses (FMDV), characterized by co-circulating O, A, and Asia 1 serotypes, as designated by the world reference laboratory for FMD (WRL-FMD). FMDV serotype A lineage ASIA/Iran-05 is widespread in buffalos and cattle populations and was first reported in Pakistan in 2006. This lineage has a high turnover, with as many as 10 sub-lineages reported from Pakistan over the years. In this study, we reconstructed the evolutionary, demographic, and spatial history of serotype A and one of its sub-lineages, A/ASIA/Iran-05/SIS-13, prevalent in Pakistan. We sequenced nearly complete genomes of three isolates belonging to sub-lineage A/ASIA/Iran-05/SIS-13. We estimated recombination patterns and natural selection acting on the serotype A genomes. Source and transmission routes in Pakistan were inferred, and the clustering pattern of isolates of the SIS-13 sub-lineage were mapped on a tree. We hereby report nearly complete genome sequences of isolates belonging to sub-lineage A/ASIA/Iran-05/SIS-13, along with purported recombinant genomes, and highlight that complete coding sequences can better elucidate the endemic history and evolutionary pressures acting on long-term co-circulating FMDV strains.

Molecular detection of foot and mouth disease virus serotype A in goats (Capra hircus)

Foot and mouth disease (FMD) is a highly contagious, transboundary, and re-emerging viral disease that mostly affects cloven-hoofed animals specially cattle, goats, swine, sheep, etc. resulting in significant economic losses. Despite regular vaccination, outbreaks of the disease have become a yearly occurrence across the country. RT-PCR was used to determine the prevalence and molecular detection of serotype A of FMDV in clinically affected goats in Goat research farm, BLRI, Savar, Dhaka, Bangladesh during August 2018. A total of 9 samples were taken from 1 to 2 years old Black Bengal goats suspected to have FMD. FMDV was detected in 55.56% (5/9) of the suspected samples using RT-PCR. The serotype of positive samples was also determined using gsRT-PCR. However, FMDV serotype A was prevalent in 100% (5) positive samples. Additionally, considering the age, the prevalence of confirmed FMD outbreak was 40% (2), 40% (2), and 20% (1) at the age of 1, 1.5 and 2 years, respectively. It was found that young goats are more susceptible to FMDV than adults. However, it can be concluded that suspected goats were infected with FMDV serotype A and trivalent FMD vaccine is suggested for prevention and control of FMD outbreak. Asian J. Med. Biol. Res. 2022, 8 (1), 30-36

Genetic variation and evolution of foot–and–mouth disease virus serotype A in relation to vaccine matching

Foot–and–mouth disease (FMD) is a severe, highly contagious viral disease that affects a wide variety of domestic and wild cloven-hoofed animals. FMD vaccines can play a vital role in disease control and are very widely used globally each year. However, due to the diversity of FMDV, the choice of FMD vaccine is still a huge challenge. In this study, 45 FMDV/A isolates were phylogenetically categorized into three topotypes: ASIA (n = 31), AFRICA (n = 10), and EURO–SA (n = 4). Three sera collected from vaccinated cattle with FMDV A22/IRQ/24/64, A/IRN/05, and A/ARG/01 were used to evaluate their antigenic relationship (r1) with the field isolates. The IRQ/24/64 serum demonstrated a 39% (17/44) match (r1 ≥ 0.3) to the field isolates, whereas IRN/05 serum and ARG/01serum showed an 18% (8/44) and a 2% (1/44) match (r1 ≥ 0.3) to the field isolates, respectively. The A22/IRQ/24/64 matched with isolates mainly from topotype ASIA, with limited cross–topotype match with isolates from topotypes AFRICA and EURO–SA. However, the A/IRN/05 did not show a cross–topotype match with topotype AFRICA isolates and A/ARG/01 failed to match any isolates from topotypes ASIA and AFRICA. After analyzing the amino acid variation of the known antigenic sites of 45 strains of FMDV/A, it was found that together antigenic sites 1 and 3 contributed about 71% of the amino acid changes to the vaccine evaluated. Based on the capsid sequences, the FMDV/A evolved unequally among topotypes. The topotypes of ASIA and AFRICA evolves faster than that of EURO–SA. The FMDV/A continues to show a high level of genetic diversity driven by a high substitution rate, purifying selection, and positive selection concentrated on antigenic sites or near antigenic sites. The current research shows the challenges of the FMDV/A vaccine selection and emphasizes the importance of continuous monitoring of antigenic evolution for the selection of effective vaccines.

Differentiation of Foot and Mouth Disease Virus Serotypes in Animals in High-Risk Zones of Georgia

Foot and Mouth Disease (FMD) is the most important economic threat to the livestock industry. Outbreaks of FMD can have a devastating impact on livestock production and trade, resulting in significant economic losses in the agricultural sector. As a result, vaccination and containment programs have been implemented internationally to minimize the spread of FMDV. The national vaccination program has been implemented in Georgia since 2012, vaccinating Large Ruminants (LR) and Small Ruminants (SR) with trivalent (A, O, Asia1) vaccine twice annually. However, active seromonitoring surveillance still shows a high seroprevalence of the disease, indicating virus circulation. In this study we attempted to estimate the prevalence of different FMDV serotypes in various risk zones within Georgia. A total of 4991 small and large ruminants were tested for the presence of FMDV nonstructural proteins (NSP) in the blood, and the exact serotypes of positive animals were further investigated through structural protein (SP) based assays. The results show that significant percentages (6.6%) of vaccinated animals were affected by FMD, and those positive animals are usually affected by more than one FMDV serotype. As such, our data call upon a stricter vaccination and monitoring program for FMDV in Georgia, especially considering that due to the geographic location of Georgia, the presence of FMD can have significant impact on transit and can be a threat for other countries as well.

Sero-detection of Foot and Mouth Disease Virus Serotypes A and O in One-humped Camels (Camelus dromedarius) in the Middle of Iraq

Sero-detection of Foot and Mouth Disease Virus Serotypes A and O in One-humped Camels (Camelus dromedarius) in the Middle of Iraq

Differential antibody responses to the major antigenic sites of FMD virus serotype O after primo-vaccination, multiply-vaccination and after natural exposure

Foot and mouth disease (FMD) virus serotype O is the predominant cause of FMD outbreaks in several regions of the world including India. Five independent neutralizing antigenic sites have been identified on the capsid surface of FMD virus serotype O. The relative importance of these neutralizing sites in eliciting antibody responses in the polyclonal sera collected from un-infected vaccinated (both primo and multiply-vaccinated) and naturally infected cattle populations were determined through a combination of reverse genetics and serology. The known critical amino acid residues present on the five antigenic sites of FMD virus serotype O Indian vaccine strain O IND R2/1975 were mutated through site-directed mutagenesis. The mutant viruses were rescued in cell-culture and analyzed through virus-neutralization assays along with parent virus using the polyclonal sera collected from three groups of cattle. In the polyclonal sera from primo-vaccinated cattle, significantly higher level of antibodies were directed towards antigenic site 2. In contrast, in polyclonal sera from multiply vaccinated animals, both antigenic sites 1 and 2 were equally important. In case of naturally infected animals, antibody responses were elicited against all the five antigenic sites. Although a drop in neutralization titres was observed for all the mutants, in one instance, increase in titre was noticed for a site 3 mutant. The findings from this study extend our knowledge on the antibody immunodominace following FMDV vaccination and infection, and may improve our strategies for vaccine strain selection and rational vaccine design.

English

English

Molecular diagnosis and genetic relationship of foot and mouth disease virus serotype Asia1/Basne/Sul/2015

Foot and Mouth Disease (FMD) is the most economically important viral-induced livestock disease worldwide. From April to May of 2015, tongue epithelial tissue samples were collected from 36 cattle in six villages, which share the border with Iran. Samples were screened using RT-PCR to amplify a conserved region in the VP1 gene, and phylogenetic tree analysis was performed based on the VP1 nucleotide sequence results. Furthermore, the nucleotide sequence was converted to an amino acid sequence in order to detect similarities between the studied samples and those previously published in GenBank (NCBI). Epidemically, based on the amino acid residues, genetic similarity, and amino acid substitutions, the VP1 nucleotide sequences were determined to be close to a novel group, group VII, with 94% identity. The VP1 amino acid sequence analysis revealed a close relationship to the Asia/BAL/PAK/iso-2/2011 isolate (Accession no. JX435109), with 95.7% identity, which is not significantly different. Analysis of the studied samples revealed that the FMDV serotype Asia1 causing the outbreak in the Basne district belonged to group VII, which was introduced from the Balochistan province of Pakistan through illegal movement of animals from this region.

Molecular and Genetic Characterization of Newly Circulating Foot and Mouth Disease Virus (FMDV) Serotype SAT2 in Egypt during 2018 and Early 2019

Molecular and Genetic Characterization of Newly Circulating Foot and Mouth Disease Virus (FMDV) Serotype SAT2 in Egypt during 2018 and Early 2019

Genetic and antigenic relationship of foot–and–mouth disease virus serotype O isolates with the vaccine strain O1/BFS

Foot–and–mouth disease serotype O viruses (FMDV/O) are responsible for the most outbreaks in FMD endemic countries. O1/BFS is one of the recommended FMD/O vaccine strains by World Reference Laboratory for FMD. In the current study, FMDV/O1 BFS vaccine strain and serotype O field isolates (45) were analyzed phylogenetically and antigenically to gain more insight into the genetic and antigenic characteristics of the vaccine strain and field isolates.O1/BFS showed similarity with 89% of the field isolates using a virus neutralization test (VNT). The P1 region encoding the FMDV capsid was sequenced and analysed for 46 strains of FMDV/O. Phylogenetic analysis showed these viruses originated from five continents and covered eight of 11 reported topotypes. Five isolates that demonstrated low antigenic similarities with O1/BFS were analyzed for their antigenic variation at the known neutralizing antigenic sites. Three of the five isolates demonstrated unique amino acid substitutions at various antigenic sites. No unique amino acid substitutions were observed for the other two unmatched isolates. Positively selected residues were identified on the surface of the FMD virus capsid supporting that it is important to continuously monitor field isolates for their antigenic and phenotypic changes.In conclusion, the vaccine strain O1/BFS is likely to confer protection against 89% of the 45 FMDV/O isolates based on VNT. Thus O1/BFS vaccine strain is still suitable for use in global FMD serotype O outbreak control. Combining data from phylogenetic, molecular and antigenic analysis can provide improvements in the process of vaccine selection.

Five BoLA-DRB3 genotypes detected in Egyptian buffalo infected with Foot and Mouth disease virus serotype O.

Foot and Mouth disease (FMD) is a contagious disease leads to economically loss in livestock production all over the world. This serious disease is caused due to the infection of the animal with a single-stranded RNA virus (FMDV). This study aimed to investigate the genetic polymorphism of BoLA-DRB3 gene in Egyptian buffalo as a candidate genetic marker included in multi-factorial process of FMD resistance/susceptibility. Also this work aimed to genetically characterization and serotyping of circulating FMD virus in Egypt during 2016.For serotyping of FMDV, RT-PCR was used for FMDV-positive samples and the results declared the presence of serotype O in all tested animals. The sequence analysis of FMDV samples revealed five different patterns for the detected serotype O which were submitted to GenBank under the accession Nos.: MG017361–MG017365.The 302-bp amplified fragments from BoLA-DRB3 exon 2 were digested with HaeIII endonuclease and the results showed that the presence of five BoLA-DRB3 genotypes, among them the genotype AA might be associated with FMD-resistance (P < 0.01). On the other hand, genotype AC could be correlated with susceptibility (P < 0.01) to FMD in Egyptian buffaloes where it was absent in resistant group. The five detected genotypes of BoLA-DRB3 exon 2 were submitted to GenBank with the accession Nos.: MF977316–MF977320. In conclusion, our findings suggested that the detection of different BoLA-DRB3 genotypes may be has a promising role for raising the resistance of Egyptian buffalo against FMDV especially serotype O which is prevalent in Egypt with preferring genotype AA.

Development and evaluation of Reverse Transcription-Polymer-ase Chain Reaction (RT-PCR) assay for differentiation of Foot and Mouth Disease virus serotype O lineages circulating in India

Development and evaluation of Reverse Transcription-Polymer-ase Chain Reaction (RT-PCR) assay for differentiation of Foot and Mouth Disease virus serotype O lineages circulating in India