Inactivated Foot-and-mouth Disease Vaccines Research Articles

Determination of Optimal Antigen Yield and Virus Inactivation Conditions for the Production of the Candidate Foot-and-Mouth Disease Recombinant Vaccine Strain Asia1 Shamir-R in a Bioreactor.

Since the foot-and-mouth disease (FMD) outbreak in South Korea in 2010-2011, vaccination policies utilizing inactivated FMD vaccines composed of types O and A have been implemented nationwide. However, because type Asia1 occurred in North Korea in 2007 and intermittently in neighboring countries, the risk of type Asia1 introduction cannot be ruled out. This study evaluated the antigen yield and viral inactivation kinetics of the recombinant Asia1 Shamir vaccine strain (Asia1 Shamir-R). When Asia1 Shamir-R was proliferated in shaking flasks (1 L), a 2 L bioreactor (1 L), and a wave bioreactor (25 L), the antigen yields were 7.5 μg/mL, 5.2 μg/mL, and 3.8 μg/mL, respectively. The optimal FMDV inactivation conditions were 2 mM BEI at 26 °C and 1.0 mM BEI at 37 °C. There was no antigen loss due to BEI treatment, and only a decrease in antigen levels was observed during storage. The sera from pigs immunized with antigen derived from a bioreactor exhibited a neutralizing antibody titer of approximately 1/1000 against Asia1 Shamir and Asia1/MOG/05 viruses; therefore, Asia1 Shamir-R is expected to provide sufficient protection against both viruses. If an FMD vaccine production facility is established, this Asia1 Shamir-R can be employed for domestic antigen banks in South Korea.

Porcine interferon-α linked to the porcine IgG-Fc induces prolonged and broad-spectrum antiviral effects against foot-and-mouth disease virus

Foot-and-mouth disease (FMD) is an economically important disease, and the FMD virus (FMDV) can spread rapidly in susceptible animals. FMD is usually controlled through vaccination. However, commercial FMD vaccines are only effective 4–7 days after vaccination. Furthermore, FMDV comprises seven serotypes and various topotypes, and these aspects should be considered when selecting a vaccine. Antiviral agents could provide rapid and broad protection against FMDV. Therefore, this study aimed to develop a fusion protein of consensus porcine interferon-α and Fc portion of porcine antibody IgG (poIFN-α-Fc) using a baculovirus expression system to develop a novel antiviral agent against FMDV. We measured the antiviral effects of the poIFN-α-Fc protein against FMDV and the enhanced duration in vitro and in vivo. The broad-spectrum antiviral effects were tested against seven FMDV serotypes, porcine reproductive and respiratory syndrome virus (PRRSV), and bovine enterovirus (BEV). Furthermore, the early protective effects and neutralizing antibody levels were tested by co-injecting poIFN-α-Fc and an FMD-inactivated vaccine into mice or pigs. Sustained antiviral effects in pig sera and mice were observed, and pigs injected with a combination of the poIFN-α-Fc and an inactivated FMD vaccine were protected against FMDV in a dose-dependent manner at 2- and 4-days post-vaccination. In addition, combined with the inactivated FMD vaccine, poIFN-α-Fc increased the neutralizing antibody levels in mice. Therefore, poIFN-α-Fc is a potential broad-spectrum antiviral and adjuvant candidate that can be used with inactivated FMD vaccines to protect pigs against FMDV.

Bestatin, A Pluripotent Immunomodulatory Small Molecule, Drives Robust and Long-Lasting Immune Responses as an Adjuvant in Viral Vaccines.

An inactivated whole-virus vaccine is currently used to prevent foot-and-mouth disease (FMD). Although this vaccine is effective, it offers short-term immunity that requires regular booster immunizations and has several side effects, including local reactions at the vaccination site. To address these limitations, herein, we evaluated the efficacy of bestatin as a novel small molecule adjuvant for inactivated FMD vaccines. Our findings showed that the FMD vaccine formulated with bestatin enhanced early, intermediate-, and particularly long-term immunity in experimental animals (mice) and target animals (pigs). Furthermore, cytokines (interferon (IFN)α, IFNβ, IFNγ, and interleukin (IL)-29), retinoic acid-inducible gene (RIG)-I, and T-cell and B-cell core receptors (cluster of differentiation (CD)28, CD19, CD21, and CD81) markedly increased in the group that received the FMD vaccine adjuvanted with bestatin in pigs compared with the control. These results indicate the significant potential of bestatin to improve the efficacy of inactivated FMD vaccines in terms of immunomodulatory function for the simultaneous induction of potent cellular and humoral immune response and a long-lasting memory response.

Preventive effects of quercetin against foot-and-mouth disease virus in vitro and in vivo by inducing type I interferon.

Foot-and-mouth disease (FMD) is an acute contagious infectious disease that affects cloven-hoofed animals. Although current emergency FMD vaccines only take effect 7 days after vaccination, antiviral agents, such as quercetin, which is a common flavonoid, could reduce the spread of FMD virus (FMDV) during outbreaks. We investigated the in vitro and in vivo antiviral effects of quercetin against FMDV. Analysis of viral copy numbers showed that quercetin had a dose-dependent inhibitory effect on FMDV at concentrations between 19.5 and 1,250 μM in porcine cells. In addition, we observed a quercetin-induced interferon (IFN)-α protein and interferon-stimulated gene (ISG) upregulation in swine cells. Enzyme-linked immunosorbent assay of sera revealed that quercetin induces the production of IFN-α, IFN-β, IFN-γ, interleukin (IL)-12, and IL-15 in mice. Inoculation of mice with quercetin or a combination of quercetin with an inactivated FMD vaccine enhanced both the survival rate and neutralizing antibody titer. Therefore, we suggest the use of quercetin as a novel and effective antiviral agent for controlling FMDV infection; however, further investigation of its application in livestock is required.

Investigation of the effect of mutual vaccination with pest des petits ruminants and polyvalent foot and mouth disease vaccines on the immune response of sheep.

Pest des petits ruminants (PPRs) and foot and mouth disease (FMD) are two viral infectious diseases affecting sheep dramatically causing great economic losses. Therefore, attention should be directed toward their control, especially through the application of well-designed vaccination schedules with specific potent vaccines. Determination of the possibility of sheep vaccination with PPR and FMD vaccines in a mutual schedule. Different groups of sheep have vaccinated with live attenuated PPR vaccine and inactivated polyvalent FMD vaccine in a mutual manner (one before the other at weekly intervals or simultaneously) followed by monitoring of the induced immunity to both vaccines using serum neutralization test (SNT) and enzyme linked immune sorbent assay (ELISA). SNT and ELISA revealed that there was no antagonizing effect of any vaccine on the immune response to the mutual vaccination of sheep to the other where the obtained antibody titers in single vaccinated sheep groups were similar to those in the simultaneous vaccinated group. Simultaneous vaccination of sheep with PPR and polyvalent FMD vaccine is of applicable benefit saving time, effort, and stress factors on the animals.

Ultrasound monitoring of the regenerative treatment of Biceps Tendonitis and Tenosynovitis in dogs by stem cells injections

Foot-and-mouth disease (FMD) and bluetongue disease (BT) are two of the most devastating viral diseases affecting domestic and wild ruminants. They cause significant economic losses, and vaccination is the most important method for combating both FMD and BT. Simultaneous vaccinations are crucial for animal welfare as well as labor and cost savings. No reports were found on the simultaneous administration of FMD and BT vaccines to sheep. This study evaluated the effect of the simultaneous administration of an inactivated FMD vaccine and an attenuated BT vaccine on the humoral immune response to both diseases. A total of 75 sixmonth- old Akkaraman-Kıvırcık crossbred sheep that did not have any vaccination history were used as research material. Before the experiment, the animals were tested for the absence of FMD and BT antibodies. The sheep were divided into three experimental groups and vaccinated with an FMD vaccine alone, a BT vaccine alone, and both. Blood samples were taken on days 30 and 60 post-vaccination. The blood samples were evaluated using liquid-phase blocking ELISA (LPBE) and virus neutralization test (VNT) for FMD antibodies, as well as competitive ELISA (c-ELISA) and VNT for BT antibodies. The study revealed that differences between the FMD and BT antibody titers of the group vaccinated simultaneously and the FMD and BT antibody titers obtained from the groups vaccinated separately were not statistically significant (p > 0.05). It was concluded that the antibody response these diseases was not altered when the FMD and BT vaccines were administered together.

Efficacy of a Novel Multiepitope Vaccine Candidate against Foot-and-Mouth Disease Virus Serotype O and A.

Foot-and-mouth disease (FMD) is a highly contagious and economically devastating disease in cloven-hoofed animals. To prevent the spread of FMD virus (FMDV), traditional inactivated vaccines are used to immunize susceptible animals in disease-endemic countries. However, the inactivated FMD vaccine has several limitations, including safety concerns. To overcome these limitations, subunit proteins have been studied as alternative vaccine candidates. In this study, we designed two multiepitope recombinant proteins (OVM and AVM) containing antigenic sites (residue of VP1 132-162 and residue of VP1 192-212) of three topotypes of FMDV serotype O or three topotypes of FMDV serotype A. Each recombinant protein was efficiently expressed in Escherichia coli with high solubility, and the immunogenicity and protective efficacy of the proteins as FMD vaccine candidates were evaluated. The results showed that OVM and AVM emulsified with ISA201 adjuvant induced effective antigen-specific humoral and cell-mediated immune responses and successfully protected mice from O/Jincheon/SKR/2014, O/VET/2013, and A/Malaysia/97 viruses. In addition, intramuscular immunization of pigs with the OVM and AVM emulsified with ISA201 elicited effective levels of neutralizing antibodies to the viruses with homologous epitopes. Importantly, OVM-AVM emulsified with CAvant®SOE-X adjuvant conferred 100% protection against the O/Jincheon/SKR/2014 virus with homologous residues and 75% protection against A/SKR/GP/2018 with heterologous residues. The results presented in this study suggest that the combination of OVM and AVM protein with an effective adjuvant could yield an effective and safe vaccine candidate for the prevention and control of foot-and-mouth disease. In addition, our results provide a vaccine platform that can safely, cost-efficiently, and rapidly generate protective vaccine candidates against diverse FMDVs.

Foot-and-mouth disease virus: DNA aptamer selection for the 3ABC protein

Foot-and-mouth disease (FMD) is a devastating livestock disease caused by foot-and-mouth disease virus (FMDV), a member of the Picornaviridae family. The 3ABC is a non-structural protein of FMDV, produced during viral replication and absent from inactivated FMD vaccines. Nucleic acid aptamers are DNA or RNA oligonucleotides capable of binding with high specificity and affinity to a molecular target. The aim of this study was to obtain DNA aptamers specific for 3ABC protein with a view of their application in the FMD diagnosis.Aptamers are usually obtained through SELEX (Systematic Evolution of Ligands by EXponential enrichment) procedure. In this study, an aptamer (termed FMDV1) was selected by a variation of this technique called Capillary Electrophoresis SELEX (CE-SELEX). The FMDV1 aptamer showed high binding affinity to the 3ABC protein with Kd value in the nano molar range: 22.69 ± 1.79 nM. The FMDV1 aptamer binding to 3ABC was significantly higher when compared with the BSA protein, used as control, demonstrating its specificity.

BacMam Expressing Highly Glycosylated Porcine Interferon Alpha Induces Robust Antiviral and Adjuvant Effects against Foot-and-Mouth Disease Virus in Pigs.

Foot-and-mouth disease (FMD) is an acute contagious disease that affects cloven-hoofed animals and has severe global economic consequences. FMD is most commonly controlled by vaccination. Currently available commercial FMD vaccines contain chemically inactivated whole viruses, which are thought to be slow acting as they are effective only 4 to 7 days following vaccination. Hence, the development of a novel rapid vaccine or alternative measures, such as antiviral agents or the combination of vaccines and antiviral agents for prompt FMD virus (FMDV) outbreak containment, is desirable. Here, we constructed a recombinant baculovirus (BacMam) expressing consensus porcine interferon alpha (IFN-α) that has three additional N-glycosylation sites driven by a cytomegalovirus immediate early (CMV-IE) promoter (Bac-Con3N IFN-α) for protein expression in mammalian cells. Bac-Con3N IFN-α expressing highly glycosylated porcine IFN-α protein increased the duration of antiviral effects. We evaluated the antiviral effects of Bac-Con3N IFN-α in swine cells and mice and observed sustained antiviral effects in pig serum; additionally, Bac-Con3N IFN-α exhibited sustained antiviral effects in vivo as well as adjuvant effects in combination with an inactivated FMD vaccine. Pigs injected with a combination of Bac-Con3N IFN-α and the inactivated FMD vaccine were protected against FMDV at 1, 3, and 7 days postvaccination. Furthermore, we observed that in combination with the inactivated FMD vaccine, Bac-Con3N IFN-α increased neutralizing antibody levels in mice and pigs. Therefore, we suggest that Bac-Con3N IFN-α is a strong potential antiviral and adjuvant candidate for use in combination with inactivated FMD vaccines to protect pigs against FMDV. IMPORTANCE Early inhibition of foot-and-mouth disease (FMD) virus (FMDV) replication in pigs is highly desirable as FMDV transmission and shedding rates are higher in pigs than in cattle. However, commercial FMD vaccines require at least 4 to 7 days postvaccination (dpv) for protection, and animals are vulnerable to heterologous viruses before acquiring high antibody levels after the second vaccination. Therefore, the development of antiviral agents for use in combination with FMD vaccines is essential. We developed a novel antiviral and immunostimulant, Bac-Con3N IFN-α, which is a modified porcine IFN-α-expressing recombinant baculovirus, to improve IFN stability and allow its direct delivery to animals. We present a promising candidate for use in combination with inactivated FMD vaccines as pigs applied to the strategy had early protection against FMDV at 1 to 7 dpv, and their neutralizing antibody levels were higher than those in pigs administered the vaccine only.

The effects of simultaneous foot-and-mouth disease and Escherichia coli vaccination on the immunity of pregnant cows and their calves

Fighting against infectious diseases with almost 18 million cattle spread over a wide geographical area is troublesome economically and physically. The administration of different vaccines simultaneously during the foot-and-mouth disease (FMD) vaccination campaign will help reduce both the vaccine stress of animals and the field veterinarians’ workload. Diarrhea due to Escherichia coli (E. coli) is a significant problem and a major cause of mortality in calves, and thus, simultaneous application of FMD and E. coli vaccines to dams would prove efficient in field conditions. This study aimed to investigate the effect of simultaneous administration of four different locally produced gel and oil adjuvant inactivated E. coli vaccines with locally produced oil adjuvant inactivated FMD vaccine on neutralizing antibody levels and the properties of the antibodies in late-term pregnant dams and their calves. For this purpose, seven to eight-month-old pregnant dams (n = 146) were divided into two main groups to receive two doses (single and booster), or only single dose (no booster). FMD neutralizing antibody titers were evaluated by a virus neutralization test against serotype A, O, and Asia1. In addition, the properties of antibodies against serotype A were assessed by isotype and avidity ELISAs. E. coli antibodies were measured by the hemagglutination inhibition test. Results of the study revealed no safety problems in any dams after the vaccinations. On day 42, calves of the dams vaccinated simultaneously had higher neutralizing antibody titers against three serotypes than the calves of only FMD vaccinated dams. IgG1/IgG2 ratio was higher in single-dose groups than booster-dose groups against serotype A in calves. The avidity index was detected over the threshold value (24.5%). A positive correlation was found in the transmission of maternal antibodies from mothers to calves in simultaneous administration groups. In conclusion, during the FMD vaccine campaigns, FMD and E. coli vaccines can be simultaneously applied to dams in the 7th and 8th months of gestation. Results of this study has led to the inclusion of simultaneous application of FMD and E. coli vaccines in the ‘Animal Disease and Animal Movement Control Program-2018′ and these two vaccines have been applied in the field ever since as a solution for field veterinarians to save time and labor during vaccination.

Immunity Evaluation of an Experimental Designed Nanoliposomal Vaccine Containing FMDV Immunodominant Peptides.

Foot-and-mouth disease (FMD) is a highly contagious viral disease affecting cloven-hoofed animals. The particular virus causing FMD disease is called FMD virus and is a member of the Aphthovirus genus in the Picornaviridae family. The FMD virus has an 8500 nt long single strain positive RNAgenome with one open reading frame (ORF) trapped in an icosahedral capsid protein. This virus genome doesn't have proofreading property which leads to high mutagenesis. It has seven serotypes, including O, A, ASIA, SAT1, SAT2, and C serotypes, as well as many subtypes. Iran is an endemic region for foot-and-mouth disease. Vaccination of susceptible animals with an inactivated whole-virus vaccine is the only way to control the epidemic in many developing countries. Today, conventionally attenuated and killed virus vaccines are being used worldwide. In Iran, animals have been vaccinated every 105 days with an inactivated FMD vaccine. Although commercially available FMD vaccines are effective, they provide short-term immunity requiring regular boosters. A new FMD vaccine is needed to improve immunization, safety, and long-term immune responses. A synthetic peptide vaccine is one of the safe and important vaccines. Peptide vaccine has low immunogenicity, requiring strong adjuvants. Nanoliposomes can be used as new adjuvants to improve immune response. In the current study, nanoliposomal carriers were selected using Dimyristoylphosphatidylcholine (DMPC), dimyristoyl phosphoglycerol (DMPG), and Cholesterol (Chol) as an adjuvant containing two immunodominant synthetic FMDV peptides. The liposomal formulations were characterized by various physicochemical properties. The size, zeta potential, and encapsulation efficiency were optimized, and the obtained nanoliposome was suitable as a vaccine. The efficacy of vaccines has been evaluated in guinea pigs as animal models. Indirect ELISA was used to detect FMDV-specific IgG. The obtained results indicated that although antibody titer was observed, the amount was lower compared to the groups that received inactivated virus-containing liposomes. In addition, the results showed that liposome was an appropriate adjuvant, compared to other adjuvants, such as Alum and Freund, and can act as a depot and induce an immune response.

The Potential Adjuvanticity of CAvant®SOE for Foot-and-Mouth Disease Vaccine.

Foot-and-mouth disease (FMD) is a notifiable contagious disease of cloven-hoofed mammals. A high potency vaccine that stimulates the host immune response is the foremost strategy used to prevent disease persistence in endemic regions. FMD vaccines comprise inactivated virus antigens whose immunogenicity is potentiated by immunogenic adjuvants. Oil-based adjuvants have clear advantages over traditional adjuvant vaccines; however, there is potential to develop novel adjuvants to increase the potency of FMD vaccines. Thus, we aimed to evaluate the efficacy of a novel water-in-oil emulsion, called CAvant®SOE, as a novel vaccine adjuvant for use with inactivated FMD vaccines. In this study, we found that inactivated A22 Iraq virus plus CAvant®SOE (iA22 Iraq-CAvant®SOE) induced effective antigen-specific humoral (IgG, IgG1, and IgG2a) and cell-mediated immune responses (IFN-γ and IL-4) in mice. Immunization of pigs with a single dose of iA22 Iraq-CAvant®SOE also elicited effective protection, with no detectable clinical symptoms against challenge with heterologous A/SKR/GP/2018 FMDV. Levels of protection are strongly in line with vaccine-induced neutralizing antibody titers. Collectively, these results indicate that CAvant®SOE-adjuvanted vaccine is a promising candidate for control of FMD in pigs.

Effect of Vegetable Oils as Adjuvants on Immune Response to Polyvalent Foot and Mouth Disease Inactivated Vaccine

Foot and mouth disease (FMD) is one of the most important viral diseases in Egypt and the main way for its control is sufficient vaccination. A vaccine that could improve early and long-lasting immunity by selecting the best adjuvants is the main target for veterinarians. In this study, different formulae from polyvalent inactivated FMD vaccine were prepared using different vegetable oils (Peanut oil, Olive oil, and Sunflower oil) supplemented with Ginseng saponin and compared with locally used Montanide ISA206 as alternative adjuvants. Evaluation of such formulae was carried out through the international quality control protocol for vaccine evaluation, vaccination of calves groups to follow up their cell-mediated immunity using lymphocytic proliferation assay and level determination of interleukine-6, interleukin12 by the fourth week post-vaccination. Humeral immune response was evaluated by recording serum neutralizing antibodies' protective values by the 6th week. All the prepared vaccine formulae were found to be potent for vaccinated calves, except the Olive oil vaccine showed week performance. Our data suggest that Peanut oil and Sunflower oil supplemented with Ginseng saponin could be used as adjuvants in polyvalent FMD vaccine with comparable results to conventionally used mineral oil Montanide ISA206.

Use of an automatic needle-free injection device for foot-and-mouth disease vaccination in dairy heifers

The foot-and-mouth disease (FMD) vaccination of Thai dairy cows is routinely conducted to control and prevent the disease. In Thailand, FMD control strategies include a subcutaneous route (s.c.) vaccination with 2 ml of inactivated FMD vaccine 2–3 times a year produced by the Department of Livestock Development (DLD). A new way of vaccination was introduced in the form of an automatic needle-free injection device. This technology has several important advantages, such as requiring less animal restraint, reduced time and labour with high precision, and a consistent delivery system. Here, the effectiveness of an automatic needle-free injection (ANFI) device was evaluated in 30 dairy heifers (randomly divided into three groups, 10 in each group) in the FMD vaccine delivery. The first group was subcutaneously (s.c.) vaccinated using a conventional hypodermic needle with 2 ml of the vaccine, the second group received the same, but using a CO<sub>2</sub>-powered ANFI device, and the third group received the vaccine using the same ANFI device, but administered intradermally (i.d.) with only 1 ml of the vaccine. The blood samples collected up to 120 days post-vaccination revealed that both injection methods resulted in a similar serological response. The results suggest that the i.d. and s.c. ANFI systems are effective and safe. Moreover, the i.d. use of the ANFI enabled the possibility to half the vaccination dose with the same efficacy. Therefore, the ANFI can be used as an alternative approach for FMD vaccination by s.c. or i.d. routes in dairy cows in Thailand. Ultimately, reducing the use of restraint devices and labour will improve the vaccination for the prevention and control of FMD and may improve the cows’ welfare.

Immunological activities of the aqueous extracts of Cistanche deserticola as a polysaccharide adjuvant for inactivated foot-and-mouth disease vaccines

ABSTRACTInactivated foot-and-mouth disease (FMD) vaccines generally require adjuvants to enhance the immunogenicity. To test whether the aqueous extracts of Cistanche deserticola (AECD) could be developed into a polysaccharide adjuvant for inactivated FMD vaccines in mice. Experimental vaccines were prepared with inactivated FMD viral antigen plus AECD or inactivated FMD vaccine plus AECD to subcutaneously immunize mice. AECD developed an FMDV-specific humoral response by increasing IgG antibodies and IL-4 in CD4+ T cells and enhanced lymphocyte proliferation and a balanced Th1/Th2 response. Importantly, AECD induced Th1 type immunity by augmenting the activation of T subsets (CD4+, CD8+, CD44+) and the induction of IFN-γ in T lymphocytes, especially CTL response and neutralizing antibodies in sera. In addition, AECD maintained the higher levels of CD80, CD40, MHC-II, CD86 on CD11c+ cells and decreased the Treg frequency. Our results suggested that AECD had the potential utility as an polysaccharide adjuvant for FMD vaccines.

Sequence Analysis of Egyptian Foot-and-Mouth Disease Virus Field and Vaccine Strains: Intertypic Recombination and Evidence for Accidental Release of Virulent Virus.

In spite of annual mass vaccination programs with polyvalent inactivated vaccines, the incidence and economic impact of foot-and-mouth disease (FMD) in Egypt is high. Viruses of the A, O and SAT 2 serotypes are endemic and repeated incursions of new lineages from other countries lead to an unstable situation that makes the selection of appropriate vaccine antigens very difficult. In this study, whole genome sequencing of a 2016 serotype A isolate from Egypt revealed a recombination event with an African serotype O virus. Based on available vaccine matching data, none of the vaccines currently used in Egypt are expected to sufficiently protect against this virus or other viruses of this lineage (A/AFRICA/G-IV) circulating there since 2012. In addition to the risk of vaccine failure caused by strain mismatch, the production of inactivated FMD vaccines is dangerous if adequate biosafety cannot be maintained. Using a high-throughput sequencing protocol optimized for short nucleic acid fragments, the composition of a local inactivated vaccine was analyzed in depth. The serotype O strain identified in the vaccine was genetically identical to viruses found in recent FMD outbreaks in Egypt.

Foot-and-Mouth Disease Virus Lacking the Leader Protein and Containing Two Negative DIVA Markers (FMDV LL3B3D A24) Is Highly Attenuated in Pigs.

Inactivated whole-virus vaccines are widely used for the control of foot-and-mouth disease (FMD). Their production requires the growth of large quantities of virulent FMD virus in biocontainment facilities, which is expensive and carries the risk of an inadvertent release of virus. Attenuated recombinant viruses lacking the leader protease coding region have been proposed as a safer alternative for the production of inactivated FMD vaccines (Uddowla et al., 2012, J Virol 86:11675-85). In addition to the leader deletion, the marker vaccine virus FMDV LL3BPVKV3DYR A24 encodes amino acid substitutions in the viral proteins 3B and 3D that allow the differentiation of infected from vaccinated animals and has been previously shown to be effective in cattle and pigs. In the present study, two groups of six pigs each were inoculated with live FMDV LL3BPVKV3DYR A24 virus either intradermally into the heel bulb (IDHB) or by intra-oropharyngeal (IOP) deposition. The animals were observed for 3 or 5 days after inoculation, respectively. Serum, oral and nasal swabs were collected daily and a thorough postmortem examination with tissue collection was performed at the end of the experiment. None of the animals had any signs of disease or virus shedding. Virus was reisolated from only one serum sample (IDHB group, sample taken on day 1) and one piece of heel bulb skin from the inoculation site of another animal (IDHB group, necropsy on day 3), confirming that FMDV LL3BPVKV3DYR A24 is highly attenuated in pigs.

High-Resolution Composition Analysis of an Inactivated Polyvalent Foot-and-Mouth Disease Vaccine.

Appropriate vaccine selection is crucial in the control of foot-and-mouth disease (FMD). Vaccination can prevent clinical disease and reduces viral shedding, but there is a lack of cross-protection between the seven serotypes and their sublineages, making the selection of an adequately protective vaccine difficult. Since the exact composition of their vaccines is not consistently disclosed by all manufacturers, incompatibility of the strains used for vaccination with regionally circulating strains can cause vaccination campaigns to fail. Here, we present a deep sequencing approach for polyvalent inactivated FMD vaccines that can identify all component strains by their genome sequences. The genomes of all strains of a commercial pentavalent FMD vaccine were de novo assembled and the vaccine composition determined semi-quantitatively. The genome assembly required high stringency parameters to prevent misassemblies caused by conserved regions of the genome shared by related strains. In contrast, reference-guided assembly is only recommended in cases where the number of strains is previously known and appropriate reference sequences are available. The presented approach can be applied not only to any inactivated whole-virus FMD vaccine but also to vaccine quality testing in general and allows for better decision-making for vaccines with an unknown composition.

Development of a recombinant vaccine against foot and mouth disease utilizing mutant attenuated Listeria ivanovii strain as a live vector

The drawbacks of conventional inactivated Foot and Mouth Disease (FMD) vaccine, such as escaping of the virus during manufacture processes prompted researchers to explore novel types of vaccine to overcome these disadvantages. Listeria ivanovii (LI) is an intracellular microorganism that possesses immune-stimulatory properties, making it appropriate for use as a live bacterial vaccine vector. The Foot and mouth disease virus (FMDV) VP1 protein is the most immunogenic part of FMDV capsid, it has most of the antigenic sites for viral neutralization.The expression of antigen gene cassette in vitro was confirmed by Western blot analysis. Mice were able to eliminate LI△actAplcB-vp1 from the liver and spleen within few days revealed a safety of the candidate vaccine. Two doses of LI△actAplcB-vp1 with 14 days of interval were injected into mice. High levels of specific IgG antibodies and CD8+ and CD4+ T cells secreted cytokines including IFN-γ, TNF-α and IL-2 against FMDV-VP1 were achieved. Based on the obtained results, LI△actAplcB-vp1 candidate vaccine utilizing Listeria ivanovii as a live vector-based vaccine could enhance a specific cellular and humoral immune responses against the inserted FMDV-vp1 heterologous genes. LI△actAplcB-vp1 candidate vaccine could be a modern tool to overcome the disadvantages of the traditional inactivated FMD vaccine.

The effect of foot-and-mouth disease vaccination on early pregnancy loss in beef heifers in Argentina

The threat of Foot and Mouth Disease (FMD) in South America has global economic implications and retaining a FMD Free status under the World Organization for Animal Health (OIE) remains a top priority. In Argentina the Servicio Nacional de Sanidad y Calidad Agroalimentaria (SENASA), the national service of agri-food health and quality, requires cattle located in the Pampean region of the Salado River basin to receive two foot-and-mouth disease (FMD) vaccinations per year, which results in one vaccination coinciding with beef cattle breeding season. While the vaccination program remains necessary, there is a growing concern amongst food animal veterinarians, that the overlap of FMD vaccination with the first 35 days of the breeding season is associated with early pregnancy loss (EPL). To address this concern, a preliminary randomized controlled trial t study was conducted to investigate the risk ratio (RR) of EPL in vaccinated, pregnant Aberdeen Angus heifers. Initially (Day 0), 858 heifers underwent fixed time-AI (FTAI). Subsequently, on day 33, following pregnancy diagnosis by transrectal ultrasonography pregnant heifers (n = 311) were randomly allocated to two treatment groups. Group 1 (162 animals) received an inactivated oil emulsion FMD vaccine, and Group 2 (149 animals) received a saline injection (control). On day 51 (18 days post vaccination), pregnancy status was re-evaluated by ultrasonography. The initial pregnancy rate (PR) on Day 33 was 58% (498/858 animals). On Day 51 (18 days post vaccination), PR in Group 1 was 96.3% (156/162 animals), and in Group 2 (control) was 98.6% (147/149 animals). The EPL in Group 1 was 3.7% (6/162 animals) and in Group 2 was 1.3% (2/149 animals). The RR of EPL in Group 1, compared to Group 2, was 2.8 (95% confidence interval: 0.6–13, p-value: 0.20). With such a wide range in confidence intervals and a p value of 0.20 a larger prospective study would be necessary to establish an unequivocally statistically significant link between heifer vaccination 33 days post FTAI and an increased risk of EPL.