Timeliness of vesicular disease notification system in Brazilian foot-and-mouth disease surveillance programme.

Animal health surveillance programmes should be reliable and informative to ensure their effective implementation. As such, the regular assessment of those aiming to demonstrate the absence of disease, as well as the ability to detect outbreaks on time, is of vital importance. Several criteria make it possible to assess the performance of surveillance systems, including timeliness, which represents the speed between steps in a surveillance system. Therefore, the objective of this study was to evaluate the variability in the timeliness, within and between states, of the surveillance programme of the Brazilian Veterinary Services (BVS) for foot-and-mouth disease (FMD), for the notification of vesicular disease. A total of 14 years (2004-2017) of data relating to vesicular syndromes from the Brazilian Continental Information and Surveillance System (SivCont) were included. A categorical variable was created with four classes to group the notified vesicular processes in the SivCont, according to two criteria, the similarity of the symptoms of the diseases reported with FMD and aetiology (viral, bacterial, fungal and non-infectious). The three timeliness values (TL-1, TL-2 and TL-3) related to different portions of the FMD surveillance system were analysed as a response in a generalized linear model in which the states of Brazil were the explanatory variables. The analyses were performed separately for each notification class (FMD, vesicular stomatitis, similar symptoms and similar non-infectious symptoms) and included comparisons within and between states. The study results provide an understanding and evaluation of the timeliness of the Brazilian FMD surveillance system, thereby providing a base of knowledge from which involved agents and decision-makers can evaluate BVS and reinforce surveillance measures in the states with poorer timeliness than permitted.

Challenges for Serology-Based Characterization of Foot-and-Mouth Disease Outbreaks in Endemic Areas; Identification of Two Separate Lineages of Serotype O FMDV in Uganda in 2011.

Control of foot-and-mouth disease (FMD) in Uganda by ring vaccination largely depends on costly trivalent vaccines, and use of monovalent vaccines could improve the cost effectiveness. This, however, requires application of highly specific diagnostic tests. This study investigated outbreaks of FMD in seven Ugandan districts, during 2011, using the PrioCHECK® FMDV NS ELISA, solid-phase blocking ELISAs (SPBEs) and virus neutralization tests (VNTs), together with virological analyses for characterization of the responsible viruses. Two hundred and eighteen (218) cattle and 23 goat sera as well as 82 oropharyngeal fluid/epithelial tissue samples were collected. Some 50% of the cattle and 17% of the goat sera were positive by the PrioCHECK® FMDV NS ELISA, while SPBEs identified titres ≥80 for antibodies against serotype O FMD virus (FMDV) in 51% of the anti-NSP positive cattle sera. However, 35% of the anti-NSP positive cattle sera had SPBE titres ≥80 against multiple serotypes, primarily against serotypes O, SAT 1 and SAT 3. Comparison of SPBEs and VNTs for the detection of antibodies against serotypes O, SAT 1 and SAT 3 in 72 NSP positive cattle sera showed comparable results against serotype O (P = 0.181), while VNTs detected significantly fewer samples positive for antibodies against SAT 1 and SAT 3 than the SPBEs (P < 0.001). Detection of antibodies against serotype O was consistent with the isolation of serotype O FMDVs from 13 samples. Four of these viruses were sequenced and belonged to two distinct lineages within the East Africa-2 (EA-2) topotype, each differing from the currently used vaccine strain (EA-1 topotype). The relationships of these lineages to other serotype O viruses in the Eastern Africa region are discussed. To enhance the control of FMD in Uganda, there is need to improve the specificity of the SAT-SPBEs, perform vaccine matching and implement improved regional FMD control.

Cattle remain immunocompetent during the acute phase of foot-and-mouth disease virus infection.

Infection of cattle with foot-and-mouth disease virus (FMDV) results in the development of long-term protective antibody responses. In contrast, inactivated antigen vaccines fail to induce long-term protective immunity. Differences between susceptible species have also been observed during infection with FMDV, with cattle often developing persistent infections whilst pigs develop more severe symptoms and excrete higher levels of virus. This study examined the early immune response to FMDV in naïve cattle after in-contact challenge. Cattle exposed to FMDV were found to be viraemic and produced neutralising antibody, consistent with previous reports. In contrast to previous studies in pigs these cattle did not develop leucopenia, and the proliferative responses of peripheral blood mononuclear cells to either mitogen or third party antigen were not suppressed. Low levels of type 1 interferon and IL-10 were detected in the circulation. Taken together, these results suggest that there was no generalised immunosuppression during the acute phase of FMDV infection in cattle.

John Burns Brooksby CBE: 25 December 1914 - 17 December 1998.

John Brooksby was an outstanding veterinary virologist, who worked at the Animal Virus Disease Research Institute, Pirbright, for 40 years, for 16 of which he was Director of the Institute. He will be remembered for his contributions to the diagnosis of foot-and-mouth disease, for his discovery of four new types, for the classification of subtypes and for fundamental studies of the virus. As Deputy Director and Director he was responsible for programmes on fundamental investigations of foot-and-mouth disease virus and other viruses exotic to the UK and for the application of the results both in the UK and worldwide. His advice on the distribution and the control of foot-and-mouth disease was sought by international organizations and by individual countries and was responsible for reducing the risk of spread of disease.

Genetic variation of foot-and-mouth disease virus from field outbreaks to laboratory isolation.

Foot-and-mouth disease virus (FMDV), by nature of its RNA genome, possesses a high rate of mutation during replication. This results in extensive genetic polymorphism of virus populations in nature. The emergence of FMDV variants during replication has been reported. Genetic changes in the viral capsid protein (VP1) gene can result in amino acid changes affecting the immunodominant epitopes of FMDV. The genetic heterogeneity of FMDV in the field and the antigenic variants observed after cell culture isolation has been investigated by PCR sequencing and reactivity with monoclonal antibodies. These methods were applied to viruses causing two different outbreaks of FMD before and after replication in cell culture and in the animal host. The VP1 region of the genome was amplified by PCR and sequenced to reveal variant sequences identified after passage and to determine their presence in the original field tissue. In one case, reactivity with monoclonal antibodies was lost after passage as a result of an amino acid change in the subpopulation. These findings suggest that host cells can select specific virus genetic and antigenic subpopulations during virus isolation and propagation.

Genetic relationships between foot-and-mouth disease type Asia 1 viruses.

The sequence of 165 nucleotides at the 3' end of the 1D (VP1) gene of foot-and-mouth disease (FMD) virus was determined for 44 type Asia 1 strains isolated from throughout Asia between 1954-92. Analysis of the relationships between the virus genomes showed epidemiological links not previously evident. The possible origin of the only outbreak of FMD Asia 1 to have occurred in Europe, in Greece in 1984, was identified because the nucleotide sequence of this virus was closely-related to the sequences of those present in the Middle East between 1983-5. Variation in the region sequenced was not as great as that seen in the other FMDV serotypes and all viruses shared greater than 85% nucleotide identity. Thus all the virus isolates examined were considered to belong to a single genotype. A database of Asia 1 virus sequences has been established which will facilitate the rapid analysis of new outbreaks strains.

Analysis of mixed foot-and-mouth disease virus infections in Saudi Arabia: prolonged circulation of an exotic serotype.

Plaque purification of foot-and-mouth disease (FMD) type O viruses isolated from cattle in Saudi Arabia showed the presence of mixed serotype infections. Sixteen out of 31 samples collected between 1985 and 1991 also contained Asia 1 virus, a serotype which had previously only been isolated from a single outbreak in that country in 1980. Nucleotide sequences of the Asia 1 component of all these samples revealed little variation and showed that they were closely related to both a Russian lapinized vaccine virus strain (Asia 1/Tadzhikistan/64), and to a field isolate from Turkey (Asia 1/TUR/15/73). Although mixed FMD infections have been observed previously this is a first report of a serotype, considered to be exotic to a country, co-existing undetected for an extended period of time.

Experimental transmission of foot-and-mouth disease virus from carrier African buffalo (Syncerus caffer) to cattle in Zimbabwe.

Four female cattle and three male African buffalo (Syncerus caffer) which were free of foot-and-mouth disease (FMD) virus were held together on an island in Lake Kariba, Zimbabwe. The buffalo were experimentally infected with FMD virus type SAT2, developed generalised disease and became virus carriers. While the buffalo were in the acute phase of the disease the susceptible contact cattle did not show lesions, no virus was recovered from them and they did not develop serum antibodies. However, five months later the cattle developed severe foot-and-mouth disease. Direct nucleotide sequencing of the virus used to infect the buffalo and of the virus from the in-contact cattle showed that the two isolates were almost identical. The results suggest that in nature it is possible for the virus to be transmitted from buffalo to cattle under the influence of factors not yet defined, and that there was very little change in the nucleotide sequence of the virus during the carrier period of five months.

Analysis of sites of foot and mouth disease virus persistence in carrier cattle via the polymerase chain reaction.

This study was undertaken in order to explore possible sites of foot-and-mouth disease virus (FMDV) persistence during the carrier state. Tissue samples taken from experimentally infected animals at different times post-infection (p.i.) were examined by conventional viral isolation and the polymerase chain reaction (PCR) technique. The analysis of samples from several organs taken from 17 bovines between 3 and 270 days p.i. allowed the following conclusions: 1) Virus present in oesophageal-pharyngeal fluids (OPF) during the carrier state originates in the pharynx as shown by the detection of antisense FMDV RNA by PCR, 2) PCR is more sensitive than standard virus isolation techniques and may be used for the rapid detection of FMDV in specimens obtained during the acute stage of FMD and for identification of persistently infected cattle.

Immunogenicity of non-structural proteins of foot-and-mouth disease virus: differences between infected and vaccinated swine.

Non-structural as well as VP1 recombinant proteins of foot-and-mouth disease virus (FMDV) produced in E. coli, have been used to study the specific antibody response of infected or vaccinated swine. An analysis of sera from infected pigs, using a direct ELISA, showed that polypeptide 3ABC (spanning non-structural proteins 3A, 3B and 3C) was the most antigenic among the recombinant proteins studied and allowed specific detection of FMDV infected swine from the second week after the infection. The sensitivity of this assay was comparable to that obtained when the whole FMDV was used as ELISA antigen. Conversely, use of polypeptide 3ABC did not allow detection of significant levels of antibodies in sera from vaccinated animals. This differential pattern of ELISA reactivities offers a promising approach for the distinction of infected from vaccinated pigs. In addition, a highly specific and sensitive method of diagnosis for FMDV replication was achieved using an immunoblotting assay which detected antibodies against the 3ABC polypeptide.

Studies on foot and mouth disease in the eastern region of Abu Dhabi, United Arab Emirates.

The present paper describes the status of foot and mouth disease (FMD) in the eastern region of Abu Dhabi, United Arab Emirates, over a three-year period from 1987 to 1989. The disease was prevalent among the various livestock populations in the study area. Type O appeared to be most prevalent followed by types A and Asia 1 which were recorded at lower incidences. Type C was not recorded during the period of study. Two isolates of each serotype were tested for antigenic relatedness to each other and to a reference virus. Both type O isolates were closely related but less related to the reference virus O1 BFS (1860). The type A isolates and the reference virus A22 IRQ 24/64 showed a similar degree of close interrelatedness. In contrast, the two Asia 1 field isolates, while being closely related, showed no antigenic relationship with the reference virus Asia 1 PAK 1/54. The possible implications of these results regarding the current situation of FMD control in the United Arab Emirates and neighbouring countries are discussed.

Field and laboratory analysis of an outbreak of foot and mouth disease in Bulgaria in 1991.

In July 1991, an outbreak of foot and mouth disease (FMD) occurred near Stefan Karadjovo village in Boliarovo (south-east Bulgaria, close to the Turkish border). The virus isolated was identified in Bulgaria as serotype O and this was subsequently confirmed by the World Reference Laboratory for Foot and Mouth Disease in Pirbright (United Kingdom). Serological studies using bovine sera and monoclonal antibody analysis were made. In addition, the sequence of approximately 170 nucleotides at the 3' end of the 1D gene was determined for the field isolate and for vaccine strains used in Bulgaria. These were compared with other sequences of type O FMD viruses from outbreaks in the Middle East. Serum samples were taken from domestic animals in the region close to the outbreak and examined for anti-FMD virus antibodies to assess the extent (if any) of spread of the virus before or after the outbreak. No evidence of infection was found in these animals. The virus involved in the Bulgarian outbreak was antigenically similar to the O1 vaccine strains but probably did not originate from these strains. The virus was closely related genetically to a group of viruses isolated in the Middle East since 1987, suggesting that it may have been introduced into Bulgaria from an area in the Middle East by unidentified means.

The detection of foot-and-mouth disease virus in oesophageal-pharyngeal samples by a polymerase chain reaction technique.

A polymerase chain reaction (PCR) technique was used to detect the presence of foot-and-mouth disease virus (FMDV) in oesophageal-pharyngeal(OP) samples from experimentally infected steers. Ten-fold dilutions of OP samples were also diluted, inoculated onto lamb kidney cell cultures, and incubated overnight. The cultures that did not show overt cytopathogenic effects (CPE) of FMDV infection were frozen and thawed; both the fluid and the cell pellet were tested by the PCR technique. The PCR detected FMDV in the fluids of 57% of the cell cultures inoculated with 2-20 tissue culture infective doses-50% (TCID50) and of 33% of cell cultures inoculated with the 0.4-2 TCID50. The PCR detected FMDV in cell pellets of all cell cultures inoculated with 20 TCID50, of 71% of cell cultures inoculated with 2-20 TCID50 and of 50% of cell cultures inoculated with 0.2-2 TCID50. A diagnostic scheme using PCR and cell culture that would provide rapid and sensitive detection of FMDV in OP samples is proposed.

Detection of foot-and-mouth disease viral sequences in clinical specimens and ethyleneimine-inactivated preparations by the polymerase chain reaction.

The polymerase chain reaction method (PCR) has been applied to the diagnosis of foot-and-mouth disease viral RNA in tissues and, particularly, oesophageal-pharyngeal fluid (probang) samples from cattle. Using primer sets which corresponded to conserved regions of the VP1 sequence of the viral genome, it was possible to amplify sequences regardless of the serotype/strain of the virus. In comparison with infectivity assays, the PCR was generally more sensitive although there were a number of examples where only infectivity was detectable. In experiments with uninfected probang samples deliberately seeded with a dilution series of virus, the PCR proved to be approximately 10(4) times more sensitive than infectivity assays. This greater sensitivity was attributed, in part, to the ability of the PCR to amplify specifically from non-infectious RNA preparations. This enabled the identification, by sequencing, of viral RNA from chemically inactivated virus concentrates typical of those used for commercial vaccine production. Amplification of specific PCR products was also achieved with virus eluted from commercial vaccine, including preparations which had been stored for more than 10 years at 4 degrees C. The PCR technique is of considerable value, therefore, both as a complement to infectivity assays and as a powerful tool in vaccine-associated studies.

Maintenance of foot and mouth disease viruses in buffalo (Syncerus caffer Sparrman, 1779) in southern Africa.

Using age-related infection rates derived from serological data in available deterministic and specially developed stochastic simulation models, it has been possible to establish that the basic reproductive rates for South African Territory (SAT) type foot and mouth disease virus in buffalo (Syncerus caffer) are high. The models predict that there is a periodicity of infection within herds and possibly the population as a whole. Thus, buffalo herds are likely to be more infectious at some times than at others. However, because most infections in buffalo are inapparent, such episodes are difficult to identify. There is wide intratypic variation within the SAT type virus populations circulating in buffalo. This was determined by sequencing part of the 1 D gene of buffalo isolates and establishing antigenic profiles with neutralising monoclonal antibodies and conventional antisera.

The World Reference Laboratory for Foot and Mouth Disease: a review of thirty-three years of activity (1958-1991).

The range of activities and contributions of the World Reference Laboratory for Foot and Mouth Disease in Pirbright, Surrey, United Kingdom, from 1958 to 1991 is reviewed. The countries for which a service has been provided, the number of samples submitted for investigation and the serotypes identified are recorded. Factors which have influenced the number of samples received are outlined. The developments and improvements made in the laboratory diagnosis of vesicular virus diseases over the thirty-three-year period are described.

A review of foot and mouth disease in Nepal.

Foot and mouth disease (FMD) causes substantial economic losses to the predominantly agricultural community of the Kingdom of Nepal. FMD is endemic in the country and four of the seven serotypes of FMD virus have been isolated (O, A, C and Asia 1). The epidemiology of FMD and the factors which play a role in its prevalence and spread are outlined. The National Epidemiological Laboratory for FMD has been established in Kathmandu and its diagnostic capabilities and activities are described. The important points to be considered in the formulation of any future regional or national control programme for FMD in Nepal are discussed.

A comparison of type O foot and mouth disease virus field isolates from northern Thailand.

A survey of type O foot and mouth disease (FMD) virus isolates from northern Thailand was undertaken to determine the relationship between field viruses and the vaccine in use, and to gauge the range of antigenic variation among field viruses. Isolates were collected from the two most recent epizootics, 1986-1987 and 1989-1990, and assessed using a two-dimensional neutralisation test to determine their relationship to FMD type O1 Bangkok 1960 (O BKK/60) reference (vaccine challenge) virus. The critical r value for the survey was 0.259 and all isolates tested were found to have an r value considerably greater than this (range 0.66 to 0.80). The results showed close antigenic relationships between the isolates and the reference virus, and indicated a relatively small range of antigenic variation between the isolates.

A pathogenesis study of foot-and-mouth disease in cattle, using in situ hybridization.

Eight calves were exposed in an aerosol chamber to nebulized foot-and-mouth disease virus. Two control animals were exposed in a similar manner to cell culture media only. Animals were euthanized at intervals and various tissues examined by in situ hybridization using a biotinylated RNA probe corresponding to a portion of the viral gene coding for the polymerase enzyme. By this technique large amounts of viral nucleic acid were found in coronary band, interdigital cleft and tongue as early as six hours postexposure, indicating a very rapid delivery from the portal of entry to the predilection sites for lesion development. This occurred well before the onset of viremia which by virus isolation was not detectable until 30 hours postexposure. The in situ hybridization signal in these tissues decreased in intensity and extent with time to focally positive areas, occasionally surrounding a vesicle. Other epidermal sites not normally thought of as sites for foot-and-mouth lesion development, such as carpus and eyelid, also had some viral nucleic acid detectable at various time intervals. In the lung by in situ hybridization, alveolar septa had viral nucleic acid early in infection (6-18 h postexposure) while later (36-96 h postexposure), the in situ hybridization signal was prominent in alveolar macrophages.

Foot-and-mouth disease virus typing by complement fixation and enzyme-linked immunosorbent assay using monovalent and polyvalent antisera.

An indirect "sandwich" enzyme-linked immunosorbent assay (ELISA) using polyvalent and monovalent antisera was compared with the 50% complement fixation (CF50) test for the detection of foot-and-mouth disease (FMD) O, A, and C virus types. ELISA was more sensitive than CF50 tests when polyvalent antisera were used for detecting the 3 types of virus in epithelial samples, whereas ELISA using monovalent antisera was the least sensitive technique. The ELISA performed with polyvalent antisera was 9 times more sensitive for detecting FMD virus than that with monovalent antisera. However, viral isolation in cell culture was the most sensitive detection system. The combined use of ELISA with polyvalent antisera and cell culture inoculations was the most effective procedure for identifying FMD virus in epithelial samples from the field.