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Immune responses of sheep to quadrivalent double emulsion foot-and-mouth disease vaccines: rate of development of immunity and variations among other ruminants.

Despite representing the majority of the world's foot-and-mouth disease (FMD)-susceptible livestock, sheep and goats have generally been neglected with regard to their epidemiological role in the spread of FMD. In the present investigations, FMD virus quadrivalent double emulsion (Montanide ISA 206) vaccines were tested in sheep. The oil adjuvant elicited a better immune response at any time than did aluminum hydroxide gel vaccine, and the response developed quicker. The animals maintained their neutralizing antibody titers at >3 log(10) for the duration of the trial (90 days). Sheep were found to be late responders to serotypes A, C, and Asia-1; a clear upward shift in titer was observed at 60 days postvaccination. However, development of the immune response to serotype O in sheep was superior to that in cattle and goats.

Adjuvants, Immunologic↗

Molecular epidemiological investigation of foot-and-mouth disease virus in Korea in 2000.

The genetic relatedness of 7 Korean type O field strains of foot-and-mouth disease virus (FMDV) in clinical specimens collected from 5 different geographic locations in 2000 was investigated. The sequence of 162 nucleotides (nt 478-639) at the 3' end of the 1D (VP1) genes was determined from amplified cDNA fragments, and subjected to the analysis for the sequence identity/divergence and phylogenetic relationship. The overall nucleotide sequence divergence among the 7 field strains was 0 to 3.8%, suggesting that they are closely related to each other. Phylogenetic analysis with the known Middle East-South Asia (ME-SA) topotype strains showed that the 7 Korean field strains formed two distinct clusters within the same lineage of the ME-SA topotype strains. Cluster 1 consisted of the strains of the primary foci of infection (Paju and Hongseong), and closely related to the strains prevailed in the Far East. Cluster 2 comprised those of subsequently affected regions (Boryeong, Yongin, and Chungju), and was further diverged from the Cluster 1. The result of phylogenetic analysis indicated that the Korean strains may have evolved from a common ancestor of the Pan Asia strains, and that at least 2 phylogenetically clustered variants within the same lineage were prevalent during the epidemic. The potential origin and sources of the virus introduction to Korea were discussed.

Animals↗

Liposome encapsulated subunit (VP1) and virion vaccines against foot-and-mouth disease.

Subunit vaccine prepared from VP1 protein of foot-and-mouth disease virus (FMDV) types 0 and Asia 1 protected guinea pigs against FMD and also induced high levels of antibody. Liposomes have been used as a safe and potent immunological adjuvant for FMD vaccines. Vaccines prepared from inactivated virus types 0 and Asia 1 encapsulated in liposomes protected guinea pigs against challenge with homologous virus and showed good antibody response in pigs on a small scale field trial.

Adjuvants, Immunologic↗

African swine fever interference with foot-and-mouth disease infection and seroconversion in pigs.

Initial oral infection of pigs with either highly virulent (L-60) or moderately virulent (DR-2) African swine fever virus (ASFV), followed in 3 days with exposure to foot-and-mouth disease virus (FMDV) (tongue inoculation and contact), failed to cause FMDV infection or seroconversion in 18 of 22 L-60-infected pigs and 13 of 34 DR-2-infected pigs. Of the 13 DR-2-infected pigs remaining free of foot-and-mouth disease (FMD), 2 pigs survived to 24 days without antibody to FMDV, despite constant contact with clinically infected pigs with FMD. Three other DR-2-infected pigs never developed FMD lesions but did develop low levels of antibody to FMDV by day 17. A group of larger pig (in which DR-2 is less virulent) infected with DR-2 and then FMDV had a rapid but suppressed immune response to FMDV. Contact pigs introduced 3 days postinoculation and inoculated with FMDV only all became infected with ASFV by contact and died. This remarkably long lasting 1-way interference with FMD infection during acute and subacute African swine fever was not anticipated. Infection with ASFV may have blocked the initial target cells (possibly dendritic cells) necessary for establishment of FMDV infection.

African Swine Fever↗

Interaction of foot-and-mouth disease virus with dendritic cells.

Despite several decades of investigation, the manner in which foot-and-mouth disease virus (FMDV) interacts with the innate and adaptive immune compartments is not completely understood. The importance of elucidating this relationship is emphasized by the inability of current FMDV vaccines to provide long-term protection and the recent outbreaks of FMDV in formerly disease-free countries. Dendritic cells (DCs) are professional antigen-presenting cells that have evolved to monitor the environment and provide a link between the innate and adaptive immune systems. Comprehending the cross-talk between DC and FMDV will provide valuable information towards understanding the host response to the virus and will aid in the design of effective tools and vaccines to block virus spread.

Animals↗

[Serotype determination of enteroviruses that cause hand-foot-mouth disease; identification of enterovirus 71 and coxsackievirus A16 from clinical specimens by using specific probe].

Coxsackievirus A16 (CA16) and enterovirus 71 (EV71) are known to be major causative agents of hand-foot-and-mouth disease prevalent in summer in Japan. Discrimination and identification of these viruses were often hampered by a nonneutralizable or nontypable virus. Therefore, a Southern blot hybridization that utilizes mixed probes specific to serotype was developed. Firstly, an approximately 650 bases spanning 5'-noncoding region to one third of VP2 including entire VP4 was amplified with a set of primers containing enterovirus common sequences and a genomic RNA as template. Secondary, the nucleotide sequences were determined using seven CA16 and eighteen EV71 strains including the standard strains, and the deduced amino acid sequences of VP4 were searched to find residues which are conserved in the same serotypes but diverged among different serotypes. Candidate positions for the mixed probes were defined at the carboxyl terminus of VP4. Thirdly, Southern blot analyses were carried out using thirty-nine enterovirus standard strains, seven CA16 isolates and sixty-six EV71 isolates previously identified by the neutralization test. The results revealed that each mixed probe exclusively bound to the homologous DNAs but not to the heterologous ones. In an attempt to determine serotypes without virus isolation, clinical specimens from hand-foot-and-mouth disease were examined. Of 78 throat swabs and 15 vesicular fluids, 71 (91.0%) and 13 (86.7%) specimens were clearly identified, indicating that the method described here offer advantages over the traditional neutralization assay: It is rapid, specific and less labor-consuming.

Amino Acid Sequence↗

Recombinant fusion protein and DNA vaccines against foot and mouth disease virus infection in guinea pig and swine.

In this study, we provide evidence that a recombinant fusion protein containing beta-galactosidase and a tandem repeat peptide of immunogenic dominant epitope of foot-and-mouth disease virus (FMDV) VP1 protein elicits high levels of neutralizing antibody and protects both guinea pigs and swine against infection. Vaccination with this fusion protein induced a FMDV-specific proliferative T-cell response and a neutralizing antibody response. The immunized guinea pigs and swine were protected against FMD type O virus infection. Two DNA plasmids expressing genes of foot-and-mouth disease were constructed. Both plasmids pBO1 and pCO1 contain a signal sequence of the swine immunoglobulin G (IgG) gene and fusion protein gene of pXZ84. The signal sequence and fusion protein gene were under the control of a metallothionein promoter in the case of the pBO1 plasmid and under the control of a cytomegalovirus immediate early promoter in the case of pCO1 plasmid. When pBO1 and pCO1 were inoculated intramuscularly into guinea pigs, both plasmids elicited a neutralizing antibody response and spleen cell proliferation increased following stimulation with FMDV antigen, but animals were not protected from viral challenge.

Animals↗

Infectious foot-and-mouth disease virus derived from a cloned full-length cDNA.

A full-length cDNA plasmid of foot-and-mouth disease virus has been constructed. RNA synthesized in vitro by means of a bacteriophage SP6 promoter inserted in front of the cDNA led to the production of infectious particles upon transfection of BHK-21 cells. These particles were also found to be highly infectious for primary bovine kidney cells as well as for baby mice. The difficulty in cloning the foot-and-mouth disease virus cytidyl tract in Escherichia coli was circumvented by joining two separate cloned parts, representing the S and L fragments of the genome, and, in a second step, inserting a dC-dG homopolymer. Homopolymeric sequences of up to 25 cytidyl residues did not lead to the production of virus. Replicons containing poly(C) tracts long enough to permit virus replication were first established in yeast cells. One of these constructs could also be maintained in E. coli and was used to produce infectious RNA in vitro. The length of the poly(C) sequence in this cDNA plasmid was 32 nucleotides. However, the poly(C) tracts of two recombinant viruses found in transfected BHK-21 cells were 60 and 80 nucleotides long, respectively. Possible mechanisms leading to the enlargement of the poly(C) tract during virus replication are discussed.

Animals↗

Immune response characteristics following emergency vaccination of pigs against foot-and-mouth disease.

Pigs were vaccinated with the emergency inactivated foot-and-mouth disease virus (FMDV) vaccine--water-in-oil-in-water emulsion with Montanide ISA206--known to protect after 3-5 days. Peripheral blood leukocyte (PBL) sub-populations did not differ between vaccinates and controls post-vaccination. There was neither lymphopenia nor inflammatory reaction. FMDV-specific antibody and T lymphocyte activity developed in the vaccinates. Virus-induced Th1-like cytokine protein and mRNA (IFNgamma and IL-2) were identified, particularly IFNgamma. Th2-like cytokine protein and mRNA (IL-4 and IL-6) were also induced in an FMDV-specific manner. IL-10 was induced by both virus and mock antigen. The current emergency FMDV vaccine induces a diverse immune defence network--innate, and both Th1-like and Th2-like responses--without adverse reactions such as lymphopenia or inflammatory responses.

Animals↗

Expansion of host-cell tropism of foot-and-mouth disease virus despite replication in a constant environment.

Foot-and-mouth disease virus (FMDV) variants adapted to BHK-21 cells showed an expanded host-cell tropism that extended to primate and human cell lines. Virus replication in human HeLa and Jurkat cells has been documented by titration of virus infectivity, quantification of virus RNA, expression of a virus-specific non-structural antigen, and serial passage of virus in the cells. Parallel serial infections of human Jurkat cells with the same variant FMDVs indicates a strong stochastic component in the progression of infection. Chimeric viruses identified the capsid as a genomic region involved in tropism expansion. These results indicate that, contrary to theoretical predictions, replication of an RNA virus in a constant cellular environment may lead to expansion of cellular tropism, rather than to a more specialized infection of the cellular type to which the virus has been adapted.

Adaptation, Physiological↗

A serological evaluation of 1979-1982 Kenyan foot-and-mouth disease type SAT 2 viruses.

Serological evaluations of foot-and-mouth disease type SAT 2 viruses isolated in Kenya between 1979 and 1982 were performed using the two-dimensional microneutralization test. Nine field isolates of epizootiological significance were compared with four vaccine viruses. The results obtained identified Tan 5/68 as the most appropriate reference vaccine virus strain since it had the broadest serological spectrum. Potent Tan 5/68 vaccines would be expected to provide adequate protection against the contemporary SAT 2 field viruses. In the case of K183/74, which also was shown to have a broad spectrum with viruses isolated in Kenya, the results show that the 1982 isolate from central Kenya was significantly divergent (r less than 1.00 at P = 0.01) and warranted tactical revaccination for its control. The study highlighted the fact that strain R1215 which had been isolated from the oesophageal-pharyngeal swabs of asymptomatic carrier cattle had a narrow serological spectrum suggesting that such viruses could be unsuitable as vaccine for the national campaign.

Animals↗

Herd demographics correlated with the spatial distribution of a foot-and-mouth disease epidemic in Buenos Aires province, Argentina.

During a recent foot-and-mouth disease epidemic in Argentina, cattle herds affected in 2001 were located mainly (69%) in Buenos Aires province. The densities of outbreaks (no. of outbreaks per km2) and cattle-demographic variables in the province were estimated using a geographical information system and kernel function. Before the epidemic officially was recognized, the density of outbreaks was correlated (rsp = 0.28-0.47) with the geographic distribution of small (< or =100 cattle), dairy and fattening herds. During the mass-vaccination campaign to control the epidemic (April-July), the density of outbreaks was most strongly correlated (rsp = 0.20-0.25) with the distribution of large (>500 cattle) and breeding herds. After the end of the mass-vaccination campaign, large herds and number of cows were most strongly correlated (rsp = 0.16-0.26) with outbreak density. These relationships might indicate that: (1) the disease spread more rapidly or was more easily detected in intensive production systems at the beginning of the epidemic; (2) vaccination and other control methods applied were less effective in large, semi-intensive production systems; (3) incomplete vaccine protection was responsible for herd outbreaks that occurred after the end of the mass-vaccination campaign.

Animals↗

Early and cost-effective identification of high risk/priority control areas in foot-and-mouth disease epidemics.

Geo-referenced data from the 2001 Uruguayan foot-and-mouth disease (FMD) epidemic were explored to assess whether spatial analysis could lead to cost-benefit based policies. Four variables were analysed: (i) location and size of 4022 individual rural land parcels, of which 574 were infected over 60 days, (ii) animal density, (iii) percentage of dairy farms per county, and (iv) road density. Each variable was categorized into two to five classes (e.g. small/medium/large) and the proportion of cases per class reported at days 1-3 of the epidemic was compared with that reported at days 4-6. A higher proportion of cases was found at days 4-6 than at days 1-3 in areas with: small and medium size land parcels, high animal density, > 20% farms specialized in dairy production, and high road density (P < 0.03 for each). Each of these classes showed a greater proportion of cases at days 7-60 than the proportion of the total territory covered by each class's area (early case concentration ratios: 1.14-1.37). Land parcel clusters were indicated by Moran's I-test (P < 0.01). A new region was constructed by intersecting the four spatial classes associated with higher proportions of cases at days 4-6. At days 7-60, this region included 50.4% of all cases and represented 30.6% of the territory under study (final case concentration ratio: 1.65). The final area per case in this region was at least 33% lower and covered at least 45% less territory than any of the four single-variable approaches. Bio-statistical, multivariate spatial analysis of early cases may greatly increase the efficiency of epidemiologic policy.

Animal Husbandry↗

Foot-and-mouth disease in the Americas: epidemiology and ecologic changes affecting distribution.

Foot-and-mouth disease(FMD) was first recorded in South America (SA) circa 1870, in Buenos Aires, Argentina, in Uruguay, and in southern Brazil as a result of the introduction of cattle from Europe during the early days of colonization. Livestock production to trade with neighboring countries was established in the La Plata Region, and the trade of livestock and products with Chile, northeastern and central western states of Brazil, to Peru, Bolivia, and Paraguay spread FMD, which reached Venezuela and Colombia in the 1950s and finally Ecuador in 1961. The traditional forms of livestock husbandry influence the diffusion and maintenance of the FMD virus (FMDV) in different areas. Cattle production in SA depends mainly on a strong relation between cattle-calf operations and fattening operations in a complementary cycle, revealing the vulnerability and susceptibility of these areas to FMDV. Understanding the relationship between time-space behavior of the disease and the forms of production defines the FMD ecosystems, a key concept to elaborating the control/eradication strategies of national FMD eradication programs, which must be modified when trade opportunities between zones of differing sanitary status change. The role of other susceptible species besides bovines, including wildlife, in maintaining and spreading FMDV has been the subject of several studies, but in SA, bovines are so far considered to determine disease presentation. Buffalo (Bubalus bubalis) have been implicated in the spread of the disease between farms in at least one case in Brazil. Sheep are almost on a par with bovine in terms of number, especially in the Southern Cone, but their role in the maintenance of infection is not considered important, possibly owing to rearing practices. Camelid populations in the Andean region do not play an important role in the maintenance of FMD, because of short persistence of infection and low population densities in these species. The importance of wildlife is not clear, but it is accepted that animals are mostly affected as a spinoff during outbreaks in domestic species. Experimentally infected capybaras (Hydrochoerus hydrochoeris hydrochoeris) showed clinical signs and infected other susceptible species, but their role in the maintenance of infection in nature is so far not clear.

Animals↗

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

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

Animals↗

Comparison of a liquid-phase blocking sandwich ELISA and a serum neutralization test to evaluate immunity in potency tests of foot-and-mouth disease vaccines.

Sera from cattle vaccinated against either foot-and-mouth disease virus (FMDV) strains A10 Holland, O1 BFS, or C1 Detmold were tested in a serum neutralization test (SNT) and a liquid-phase blocking sandwich ELISA (LBE), and the titers were compared with the results of intradermolingual challenge tests. The LBE test results were significantly more reproducible (P less than 0.005) than the SNT results. The correlation coefficients between SNT and LBE were 0.91 for FMDV strains A10 Holland and O1 BFS, and 0.82 for FMDV strain C1 Detmold (P less than 0.0005). The regression coefficient for strain A10 Holland was 0.80, for strain O1 BFS the value was 0.87, and for strain C1 Detmold it was 0.64. In probit analysis, titers at which 95% of the cattle were protected against challenge with the homologous strain were determined for the SNT and the LBE. In the SNT the 95% protection levels for strains A10 Holland were greater than or equal to 0.84, for O1 BFS greater than or equal to 1.59, and for C1 Detmold greater than or equal to 0.83. In the LBE they were greater than or equal to 1.28, greater than or equal to 1.71, and greater than or equal to 1.74, respectively. Because the SNT and the LBE are highly significantly correlated, and the LBE is more reproducible, the LBE is likely to predict protection more reliably than the SNT.

Animals↗