Search

Foot-and-mouth disease (FMD) is considered as one of the most important contagious viral diseases affecting cloven-footed animals. For effective control of FMD, immunization along with herd immunity is essential in the field conditions. To assure and track the coverage and effectiveness of the vaccination program, the serological studies are very much required after the vaccination program. The present study was aimed to investigate the prevalence of antibodies against structural proteins of FMD virus (FMDV) serotypes of O, A and Asia-1 in seven districts of western Uttar Pradesh, India, and assure the efficacy of vaccination under National Animal Disease Control Program. A total of 308 sera samples were collected from apparent healthy vaccinated cattle and buffaloes from seven districts including Amroha, Baghpat, Bareilly, Bulandsahar, Gautam Budh Nagar, Meerut and Muzaffarnagar of western Uttar Pradesh, India. Determination of antibodies against structural proteins of FMDV was carried out using solid-phase blocking enzyme-linked immunosorbent assay. The protective level of the FMDV serotypes O, A and Asia-1 included in the inactivated trivalent vaccine was 66.55, 48.05 and 47.08% in bovines, respectively. To provide the higher level of protection against the circulating FMDV, the present study recommended the thorough investigation of the immunogenic interaction between the vaccine strains and the field strains. Further investigations should also be conducted with larger sample size and across diverse geographical regions to gain a more comprehensive understanding of herd immunity. [ABSTRACT FROM AUTHOR]

Hand, foot, and mouth disease (HFMD) is a common pediatric illness mainly caused by enteroviruses, which are important human pathogens. Currently, there are no available antiviral agents for the therapy of enterovirus infection. In this study, an excellent high-content antiviral screening system utilizing the EV-A71-eGFP reporter virus was developed. Using this screening system, we screened a drug library containing 1042 natural compounds to identify potential EV-A71 inhibitors. Fangchinoline (FAN), a bis-benzylisoquinoline alkaloid, exhibits potential inhibitory effects against various enteroviruses that cause HFMD, such as EV-A71, CV-A10, CV-B3 and CV-A16. Further investigations revealed that FAN targets the early stage of the enterovirus life cycle. Through the selection of FAN-resistant EV-A71 viruses, we demonstrated that the VP1 protein could be a potential target of FAN, as two mutations in VP1 (E145G and V258I) resulted in viral resistance to FAN. Our research suggests that FAN is an efficient inhibitor of EV-A71 and has the potential to be a broad-spectrum antiviral drug against human enteroviruses. [ABSTRACT FROM AUTHOR]

Copyright of Harran University Journal of the Faculty of Veterinary Medicine / Harran Üniversitesi Veteriner Fakültesi Dergisi is the property of Harran University, Faculty of Veterinary Medicine and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Abstract After massive foot-and-mouth disease (FMD) outbreaks originated from Jincheon County from Dec. 2014 to Apr. 2015, the effectiveness of the previous FMD vaccine containing only the O1 Manisa as the O antigen, O1 Manisa + A Malaysia 97 + Asia 1 Sharmir trivalent vaccine, was questioned in South Korea, and a change in the O antigen in FMD vaccines was demanded to control the FMD caused by FMDV O/Jincheon/SKR/2014, the O Jincheon strain. Therefore, the efficacies of O1 Manisa + O 3039 bivalent vaccine and O 3039 monovalent vaccine were studied for cross-protection against heterologous challenge with the O Jincheon strain. In this study, the efficacy of the O1 Manisa + O 3039 bivalent vaccine was better than that of the O 3039 monovalent vaccine, even though the serological relationship (r 1 value) between O Jincheon and O 3039 was matched according to the OIE Terrestrial Manual. According to serological test results from vaccinated specific pathogen free pigs, virus neutralization test titers against Jincheon were good estimates for predicting protection against challenge. A field trial of the O1 Manisa + O 3039 bivalent vaccine was performed to estimate the possibility of field application in conventional pig farms, especially due to concerns about the effect of maternally derived antibodies (MDA) in field application of the FMD vaccine. According to the result of the field trial, the O1 Manisa + O 3039 bivalent vaccine was considered to overcome MDA. The results of the efficacy and field trials indicated that the O1 Manisa + O3039 vaccine could be suitable to replace previous FMD vaccines to control the FMD field situation caused by O Jincheon FMDV. [ABSTRACT FROM AUTHOR]

Small ruminants (Ovine and Caprine) constitutes a significant revenue generation component for small-scale livestock farmers across the globe especially in most developing nations including Nigeria. Viral diseases like Foot-and-mouth disease (FMD) is a major contagious disease of domestic ruminants that hampers livestock productivity in countries where the disease is still endemic. However, the role of Sheep in FMD epidemiology in Nigeria is still underestimated and poorly understood. This study was aimed at establishing the occurrence of FMD in slaughtered Sheep at Gwagwalada abattoir. One hundred and eighty (180) sera were randomly collected from slaughtered sheep and subjected to NS-Blocking ELISA test using PrioCHECK® ELISA kit at the Foot and Mouth Disease Laboratory, National Veterinary Research Institute (NVRI), Vom. The result showed that out of 180 test sera, 29 (16.0%) were evident of NSP-FMD antibodies and 151 (84.0%) were negative. This findings indicates FMD infections were within <40 days> and absence of vaccination against FMD in sheep. The consequence of co-herding of sheep as a risk factor in FMD spread and transmission within this study area could be far overwhelming. In conclusion, this preliminary finding provides information on the occurrence of FMD in all breeds of apparently healthy, non-vaccinated Rams and Ewes slaughtered in Gwagwalada abattoir. Control efforts should be put in place to prevent potential vicious circle of outbreaks that may arise from transmission of Foot and Mouth Disease Virus from slaughtered sheep in the abattoir to livestock markets and possibly herds within and around Gwagwalada Area Council. [ABSTRACT FROM AUTHOR]

Abstract Foot-and-mouth disease (FMD) is a significant endemic transboundary animal disease in Lao People's Democratic Republic (Lao PDR) and throughout the Greater Mekong Subregion (GMS). The disease has been shown to perpetuate the cycle of smallholder poverty through reduced animal production, plus limitations on market access for trading in livestock and their products. Despite significant national and multilateral efforts to control FMD over the past two decades, endemic FMD viruses (FMDVs) continue to circulate in Lao PDR. Further, the threat from new and emerging FMDVs is increasing as transboundary movements in the region intensify in response to increasing regional demand for meat. Although the economic impacts of FMD on smallholder farmers in Lao PDR are significant, studies investigating household-level risk factors for FMD are lacking. Following an outbreak of a novel FMDV (O/ME-SA/Ind2001d) in Lao PDR in 2015, a questionnaire and serological study were conducted in Naxaythong District to identify household-level risk factors associated with this outbreak, as well as endemic circulating viruses in the outbreak area. Data were analysed using a multivariable generalised estimating equation (GEE) model with a logit link function and associations were calculated as odds ratios (OR) with 95% confidence intervals (CI 95%). After adjusting for other variables, the practice of quarantining new livestock for a minimum of two weeks prior to introduction to a herd was found to be a significant protective factor during the 2015 outbreak (OR 0.225, CI 95% [0.06, 0.88], p- value 0.003). In addition, households owning one or more animals with titres to the non-structural proteins of FMDV, indicating prior infection, had 5.5 times the odds (CI 95% [6.16, 49.11], p- value <0.001) of sharing communal grazing land with neighbouring villages. These findings indicate that implementing basic on-farm biosecurity and improved husbandry measures to minimise FMDV circulation at the household level are important and reinforce the need to enhance the education of smallholder farmers in infectious disease control. [ABSTRACT FROM AUTHOR]

Abstract Foot-and-mouth disease caused by foot-and-mouth disease virus (FMDV) is one of the most highly contagious diseases of domestic animals, and leads to enormous economic loss. Currently there are two main prevention and control strategies for the disease: eradication of the infected animals in FMDV free countries, and vaccination of the susceptible animals in countries with endemic FMDV infection. Early discovery and diagnosis of the source of infection is therefore integral to the containment of FMDV. In this study, a two-step reverse transcription recombinase polymerase amplification assay combined with lateral flow detection (RPA-LFD) was developed to detect FMDV. With incubation at 38 °C, a region of the 2B gene on the FMDV genome was successfully amplified within 20 min using specific primers and a probe. The amplified RPA product can be visualized on a lateral flow dipstick. The RPA-LFD assay was highly sensitive, detecting down to 10 copies of plasmid DNA. There was no cross-reactivity with other pathogens causing vesicular lesions. In addition, 143 clinical samples were used to compare RPA-LFD with real-time PCR, with 98.6% concordance between the assays. Therefore, the developed RPA-LFD assay provides a rapid, simple, highly promising approach to be used as point-of-care diagnostics in the field. [ABSTRACT FROM AUTHOR]

Highlights • FMDV was detected by rRT-PCR in milk up to 28 days post contact challenge. • FMDV was detected in milk collected from infected farms in the field (UK 2007). • FMDV detection was possible when a milk sample was diluted up to 10−7 in negative milk. • Pooled milk has the potential to be a valuable sample type for FMDV surveillance. Abstract This study aimed to evaluate the utility of milk as a non-invasive sample type for the surveillance of foot-and-mouth disease (FMD), a highly contagious viral disease of cloven-hooved animals. Four milking Jersey cows were infected via direct-contact with two non-milking Jersey cows that had been previously inoculated with FMD virus (FMDV: isolate O/UKG/34/2001). Milk and blood were collected throughout the course of infection to compare two high-throughput real-time reverse transcription polymerase chain reaction (rRT-PCR) protocols with different RT-PCR chemistries. Using both methods, FMDV was detected in milk by rRT-PCR one to two days before the presentation of characteristic foot lesions, similar to detection by virus isolation. Furthermore, rRT-PCR detection from milk was extended, up to 28 days post contact (dpc), compared to detection by virus isolation (up to 14 dpc). Additionally, the detection of FMDV in milk by rRT-PCR was possible for 18 days longer than detection by the same method in serum samples. FMDV was also detected with both rRT-PCR methods in milk samples collected during the UK 2007 outbreak. Dilution studies were undertaken using milk from the field and experimentally-infected animals, where for one sample it was possible to detect FMDV at 10−7. Based on the peak C T values detected in this study, these findings indicate that it could be possible to identify one acutely-infected milking cow in a typical-sized dairy herd (100–1000 individuals) using milk from bulk tanks or milk tankers. These results motivate further studies using milk in FMD-endemic countries for FMD surveillance. [ABSTRACT FROM AUTHOR]

Human enteroviruses (EV) pose amajor risk to public health. This is especially so in the Asia-Pacific region where increasing numbers of hand, foot andmouth disease (HFMD) cases and large outbreaks of severe neurological disease associated with EVA71 have occurred. Despite their importance, key aspects of the emergence, epidemiology and evolution of EVs remain unclear, andmost studies of EV evolution have focused on a limited number of genes. Here, we describe the genomic-scale evolution of EV-A viruses sampled from pediatric patients withmild disease attending a single hospital in western Sydney, Australia, over an 18-month period. This analysis revealed the presence of eight viral serotypes--Coxsackievirus (CV) A2, A4, A5, A6, A8, A10, A16 and EV-A71--with up to four different serotypes circulating in any 1month. Despite an absence of large-scale outbreaks, high levels of geographical and temporalmixing of serotypes were identified. Phylogenetic analysis revealed thatmultiple strains of the same serotype were present in the community, and that this diversity was shaped bymultiple introductions into the Sydney population, with only a single lineage of CV-A6 exhibiting in situ transmission over the entire study period. Genomic-scale analyses also revealed the presence of novel and historical EV recombinants. Notably, our analysis revealed no association between viral phylogeny, including serotype, and patient age, sex, nor disease severity (for uncomplicated disease). This study emphasizes the contribution of EV-A viruses other than EV-A71 tomild EV disease including HFMD in Australia and highlights the need for greater surveillance of these viruses to improve strategies for outbreak preparedness and vaccine design. [ABSTRACT FROM AUTHOR]

Abstract: The rapid detection of foot‐and‐mouth disease virus (FMDV) is vital for the prevention of foot‐and‐mouth disease outbreaks. In this study, a polyvinylidene difluoride (PVDF)‐supported polydiacetylene (PDA) immunosensor is developed to detect FMDV, in which a polyclonal antibody against the FMDV VP1 antigen is conjugated as a specific virus‐binding module without a linker. First, a liposome‐based immunosensor is generated for the FMDV VP1 protein in the form of photopolymerized PDA colloids. Then, the VP1‐specific PDA immunosensors are modified onto PVDF strip to enable the rapid and portable detection of FMDV. Detailed analyses are performed using ultraviolet‐visible spectroscopy, dynamic light scattering, transmission electron microscopy, and scanning electron microscopy. A blue‐to‐red color transition is observed in the presence of FMDV particles, indicating the potential applications of FMDV‐specific PDA immunosensors for use in solid‐phase detection as well as via liquid‐phase liposome platforms. Thus, this work provides a rapid and simple detection for FMDV. [ABSTRACT FROM AUTHOR]

Summary: We report the laboratory analysis of 125 clinical samples from suspected cases of foot‐and‐mouth disease (FMD) in cattle and Asian buffalo collected in Pakistan between 2008 and 2012. Of these samples, 89 were found to contain viral RNA by rRT‐PCR, of which 88 were also found to contain infectious FMD virus (FMDV) by virus isolation (VI), with strong correlation between these tests (κ = 0.96). Samples that were VI‐positive were serotyped by antigen detection ELISA (Ag‐ELISA) and VP1 sequence acquisition and analysis. Sequence data identified FMDV serotypes A (n = 13), O (n = 36) and Asia‐1 (n = 41), including three samples from which both serotypes Asia‐1 and O were detected. Serotype A viruses were classified within three different Iran‐05 sublineages: HER‐10, FAR‐11 and ESF‐10. All serotype Asia‐1 were within Group VII (Sindh‐08 lineage), in a genetic clade that differs from viruses isolated prior to 2010. All serotypes O were classified as PanAsia‐2 within two different sublineages: ANT‐10 and BAL‐09. Using VP1 sequencing as the gold standard for serotype determination, the overall sensitivity of Ag‐ELISA to correctly determine serotype was 74%, and serotype‐specific sensitivity was 8% for serotype A, 88% for Asia‐1 and 89% for O. Serotype‐specific specificity was 100% for serotype A, 93% for Asia‐1 and 94% for O. Interestingly, 12 of 13 serotype A viruses were not detected by Ag‐ELISA. This study confirms earlier accounts of regional genetic diversity of FMDV in Pakistan and highlights the importance of continued validation of diagnostic tests for rapidly evolving pathogens such as FMDV. [ABSTRACT FROM AUTHOR]

Hand, foot, and mouth disease (HFMD) is a significant public health issue in China and an accurate prediction of epidemic can improve the effectiveness of HFMD control. This study aims to develop a weather-based forecasting model for HFMD using the information on climatic variables and HFMD surveillance in Nanjing, China. Daily data on HFMD cases and meteorological variables between 2010 and 2015 were acquired from the Nanjing Center for Disease Control and Prevention, and China Meteorological Data Sharing Service System, respectively. A multivariate seasonal autoregressive integrated moving average (SARIMA) model was developed and validated by dividing HFMD infection data into two datasets: the data from 2010 to 2013 were used to construct a model and those from 2014 to 2015 were used to validate it. Moreover, we used weekly prediction for the data between 1 January 2014 and 31 December 2015 and leave-1-week-out prediction was used to validate the performance of model prediction. SARIMA (2,0,0)52 associated with the average temperature at lag of 1 week appeared to be the best model (R2 = 0.936, BIC = 8.465), which also showed non-significant autocorrelations in the residuals of the model. In the validation of the constructed model, the predicted values matched the observed values reasonably well between 2014 and 2015. There was a high agreement rate between the predicted values and the observed values (sensitivity 80%, specificity 96.63%). This study suggests that the SARIMA model with average temperature could be used as an important tool for early detection and prediction of HFMD outbreaks in Nanjing, China. [ABSTRACT FROM AUTHOR]

Since 2008, outbreaks of atypical hand, foot, and mouth disease (HFMD) in children and adults have been reported worldwide. The majority of these outbreaks are caused by a new lineage of Coxsackie virus A6 (CV-A6) presenting a more severe clinical phenotype than the classical childhood HFMD caused by CVA16. Between June 2014 and January 2016, 23 cases of atypical HFMD disease presented at a Dermatology Department at a regional University Hospital in Denmark. Patients were referred by general practitioners and dermatologists with a variety of clinical diagnoses, including eczema herpeticum, vasculitis, syphilis, dermatophytid, erythema multiforme and Stevens-Johnson syndrome. Three adults and 3 children required hospitalization due to extensive skin involvement and fever. All reported patients had laboratory-confirmed enterovirus infection. This study demonstrated an upsurge in atypical HFMD caused by CV-A6 in the Region of Southern Denmark and that atypical HFMD can be difficult to diagnose clinically as it may mimic other severe skin diseases. [ABSTRACT FROM AUTHOR]

Summary: The O/Middle East‐South Asia (ME‐SA)/Ind‐2001 lineage of foot‐and‐mouth disease virus (FMDV) is endemic in the Indian subcontinent and has been reported in the Middle East and North Africa, but it had not been detected in South‐East Asia (SEA) before 2015. This study reports the recent incursions of this viral lineage into SEA, which caused outbreaks in Vientiane Capital of Lao People's Democratic Republic (PDR) in April 2015, in Dak Nong, Dak Lak and Ninh Thuan Provinces of Vietnam from May to October 2015, and in Rakhine State of Myanmar in October 2015. Disease investigations were conducted during the outbreaks and followed up after laboratory results confirmed the involvement of FMDV O/ME‐SA/Ind‐2001 sublineage d (O/ME‐SA/Ind‐2001d). Affected host species included cattle, buffalo and pig, and all the outbreaks resolved within 2 months. Animals with clinical signs were separated, and affected premises were disinfected. However, strict movement restrictions were not enforced, and emergency vaccinations were only implemented in Vientiane Capital of Lao PDR and Dak Nong and Ninh Thuan Provinces of Vietnam. Clinical samples were collected from each outbreak and examined by nucleotide sequencing of the FMDV viral protein 1 coding region. Sequence analysis revealed that the O/ME‐SA/Ind‐2001d isolates from Lao PDR and Vietnam were closely related to each other and similar to viruses previously circulating in India in 2013. Viruses collected from Myanmar were divergent from viruses of the same sublineage recovered from Lao PDR and Vietnam but were closely related to viruses present in Bangladesh in 2015. These findings imply that at least two independent introductions of O/ME‐SA/Ind‐2001d into SEA have occurred. Our study highlights the transboundary nature of foot‐and‐mouth disease (FMD) and reinforces the importance of improved FMD surveillance and promotion of safer cross‐border trade in SEA to control the risk of introduction and spread of exotic FMDV strains. [ABSTRACT FROM AUTHOR]

Summary: Effective control and monitoring of foot‐and‐mouth disease (FMD) relies upon rapid and accurate disease confirmation. Currently, clinical samples are usually tested in reference laboratories using standardized assays recommended by The World Organisation for Animal Health (OIE). However, the requirements for prompt and serotype‐specific diagnosis during FMD outbreaks, and the need to establish robust laboratory testing capacity in FMD‐endemic countries have motivated the development of simple diagnostic platforms to support local decision‐making. Using a portable thermocycler, the T‐COR™ 8, this study describes the laboratory and field evaluation of a commercially available, lyophilized pan‐serotype‐specific real‐time RT‐PCR (rRT‐PCR) assay and a newly available FMD virus (FMDV) typing assay (East Africa‐specific for serotypes: O, A, Southern African Territories [SAT] 1 and 2). Analytical sensitivity, diagnostic sensitivity and specificity of the pan‐serotype‐specific lyophilized assay were comparable to that of an OIE‐recommended laboratory‐based rRT‐PCR (determined using a panel of 57 FMDV‐positive samples and six non‐FMDV vesicular disease samples for differential diagnosis). The FMDV‐typing assay was able to correctly identify the serotype of 33/36 FMDV‐positive samples (no cross‐reactivity between serotypes was evident). Furthermore, the assays were able to accurately detect and type FMDV RNA in multiple sample types, including epithelial tissue suspensions, serum, oesophageal–pharyngeal (OP) fluid and oral swabs, both with and without the use of nucleic acid extraction. When deployed in laboratory and field settings in Tanzania, Kenya and Ethiopia, both assays reliably detected and serotyped FMDV RNA in samples (n = 144) collected from pre‐clinical, clinical and clinically recovered cattle. These data support the use of field‐ready rRT‐PCR platforms in endemic settings for simple, highly sensitive and rapid detection and/or characterization of FMDV. [ABSTRACT FROM AUTHOR]

Summary: Foot‐and‐mouth disease (FMD) is a contagious disease of cloven‐hoofed animals that causes substantial and perpetual economic loss. Apart from the contagious nature of the disease, the FMD virus can establish in a “carrier state” among all cloven‐hoofed animals. The Mithun (Bos frontalis), popularly called the “Cattle of Mountain,” is found in the geographically isolated, hilly region of north‐east India: Arunachal Pradesh, Nagaland, Manipur and Mizoram. Despite the geographical inaccessibility, infection by FMD virus has emerged as the single most devastating disease among Mithun after the eradication of rinderpest from this region. Samples from outbreaks of FMD in Mithun were analysed by sandwich ELISA, multiplex RT‐PCR (MRT‐PCR) and liquid‐phase blocking enzyme‐linked immunosorbent assay and isolated in the BHK‐21 cell line. The results indicate the presence of FMDV serotype “O.” The sequencing and molecular phylogenies have revealed close relationships in the lineage of type “O” isolates from Bangladesh. The findings will provide useful information for further research and development of a sustainable programme for the progressive control of FMD in the Mithun population. [ABSTRACT FROM AUTHOR]

Summary: An essential step towards the global control and eradication of foot‐and‐mouth disease (FMD) is the identification of circulating virus strains in endemic regions to implement adequate outbreak control measures. However, due to the high biological risk and the requirement for biological samples to be shipped frozen, the cost of shipping samples becomes one of major obstacles hindering submission of suspected samples to reference laboratories for virus identification. In this study, we report the development of a cost‐effective and safe method for shipment of FMD samples. The protocol is based on the inactivation of FMD virus (FMDV) on lateral flow device (LFD, penside test routinely used in the field for rapid immunodetection of FMDV), allowing its subsequent detection and typing by RT‐PCR and recovery of live virus upon RNA transfection into permissive cells. After live FMDV collection onto LFD strip and soaking in 0.2% citric acid solution, the virus is totally inactivated. Viral RNA is still detectable by real‐time RT‐PCR following inactivation, and the virus strain can be characterized by sequencing of the VP1 coding region. In addition, live virus can be rescued by transfecting RNA extract from treated LFD into cells. This protocol should help promoting submission of FMD suspected samples to reference laboratories (by reducing the cost of sample shipping) and thus characterization of FMDV strains circulating in endemic regions. [ABSTRACT FROM AUTHOR]

Summary: The goal of this study was to characterize the properties and duration of the foot‐and‐mouth disease (FMD) carrier state and associated serological responses subsequent to vaccination and naturally occurring infection at two farms in northern India. Despite previous vaccination of cattle in these herds, clinical signs of FMD occurred in October 2013 within a subset of animals at the farms containing juvenile‐yearling heifers and steers (Farm A) and adult dairy cattle (Farm B). Subsequent to the outbreak, FMD virus (FMDV) asymptomatic carriers were identified in both herds by seroreactivity to FMDV non‐structural proteins and detection of FMDV genomic RNA in oropharyngeal fluid. Carriers’ seroreactivity and FMDV genome detection status were subsequently monitored monthly for 23 months. The mean extinction time of the carrier state was 13.1 ± 0.2 months, with extinction having occurred significantly faster amongst adult dairy cattle at Farm B compared to younger animals at Farm A. The rate of decrease in the proportion of carrier animals was calculated to be 0.07 per month. Seroprevalence against FMDV non‐structural proteins decreased over the course of the study period, but was found to increase transiently following repeated vaccinations. These data provide novel insights into viral and host factors associated with the FMDV carrier state under natural conditions. The findings reported herein may be relevant to field veterinarians and governmental regulatory entities engaged in FMD response and control measures. [ABSTRACT FROM AUTHOR]

A new lineage of foot-and-mouth disease virus (FMDV), called A/ASIA/G-VII, emerged from the Indian subcontinent in 2015 and continues to spread in Western Asia. Currently, the distribution of viruses belonging to this lineage is defined using sequencing approaches, but other cheaper and faster diagnostic methods are urgently needed. Thus, this study describes the development and validation of a novel A/ASIA/G-VII lineage-specific real-time RT-PCR (rRT-PCR). Diagnostic sensitivity and specificity were evaluated using representative field specimens and isolates from the A/ASIA/G-VII lineage, as well as samples comprising other FMDV lineages that co-circulate in Asia (n = 54). This lineage-specific assay accurately detected all A/ASIA/G-VII samples tested (n = 29), and no detection was observed for samples belonging to other FMDV lineages (n = 25), namely A/ASIA/Sea-97, A/ASIA/Iran-05 SIS−10 , A/ASIA/Iran-05 FAR−11 , Asia1/ASIA/Sindh-08, O/CATHAY, O/ME-SA/PanAsia-2 ANT−10 , O/ME-SA/Ind-2001d, O/SEA/Mya-98. Additionally, the limit of detection was found to be at least equivalent to a pan-serotypic rRT-PCR assay. Therefore, these data indicate that this newly developed rRT-PCR assay can be applied to characterise field isolates in countries where the A/ASIA/G-VII lineage is endemic, as well as to monitor new incursions and outbreaks due to this lineage. [ABSTRACT FROM AUTHOR]

Foot and Mouth Disease Virus (FMD) is a RNA virus, member of Picornaviridae. It has seven serotypes with no cross protection among these. FMD is responsible for heavy economic losses in cattle and buffalo. So rapid and accurate diagnosis of FMD is uttermost need of this time. Several lab.techniques are developed for rapid detection of FMD but these are expensive and time consuming. Elisa, cell culture, Reverse Transcriptase (RT-PCR) and real time PCR need specific high quality equipments in lab. Whereas Lamp PCR is simple and accurate test for the rapid diagnosis of FMD. This study has been done initially with FMD known serotypes (O, A, Asia1) already in use at animal health. Viral RNAs were extracted using R Neasy Minikit (Qiagen) according to instruction manual. After extraction RNAs were eluted in 60 ul elution buffer and stored at -70C. Each strain of FMD virus was serially diluted to -5dilution in order to check the efficacy of test. Reaction was completed in 1 hr and 10 min at temperatures 600C. It was also tested in hot water bath on same temperature. Results were same in both techniques. Results showed that LAMP was more sensitive for FMD strain O (positive reaction upto -4 dilution) than others strains A and Asia -1 (positive reaction upto -3 dilutions). Further studies will be conducted to develop LAMP PCR as field test that does not require RNA extraction. [ABSTRACT FROM AUTHOR]

Foot-and-mouth disease ( FMD) is a highly contagious viral disease of cloven-hoofed animals, which can decimate the livestock industry and economy of countries previously free of this disease. Rapid detection of foot-and-mouth disease virus ( FMDV) is critical to containing an FMD outbreak. Availability of a rapid, highly sensitive and specific, yet simple and field-deployable assay would support local decision-making during an FMDV outbreak. Here we report validation of a novel reverse transcription-insulated isothermal PCR ( RT-ii PCR) assay that can be performed on a commercially available, compact and portable POCKIT™ analyser that automatically analyses data and displays '+' or '−' results. The FMDV RT-ii PCR assay targets the 3D region of the FMDV genome and was capable of detecting 9 copies of in vitro-transcribed RNA standard with 95% confidence. It accurately identified 63 FMDV strains belonging to all seven serotypes and showed no cross-reactivity with viruses causing similar clinical diseases in cloven-hoofed animals. The assay was able to identify FMDV RNA in multiple sample types including oral, nasal and lesion swabs, epithelial tissue suspensions, vesicular and oral fluid samples, even before the appearance of clinical signs. Clinical sensitivity of the assay was comparable or slightly higher than the laboratory-based real-time RT- PCR assay in use. The assay was able to detect FMDV RNA in vesicular fluid samples without nucleic acid extraction. For RNA extraction from more complex sample types, a commercially available taco™ mini transportable magnetic bead-based, automated extraction system was used. This assay provides a potentially useful field-deployable diagnostic tool for rapid detection of FMDV in an outbreak in FMD-free countries or for routine diagnostics in endemic countries with less structured laboratory systems. [ABSTRACT FROM AUTHOR]

Objective: In this study, a serological survey was conducted in unvaccinated sheep and goat populations at Pyawbwe and Meikhtila townships of Mandalay region in Myanmar to determine the seroprevalence and associated risk factors of foot and mouth disease (FMD). Materials and methods: A total of 110 sheep and 107 goat sera samples were randomly collected from Pyawbwe. Similarly, 108 sheep and 109 goat sera were collected from Meikhtila. All samples were tested for the presence of nonstructural protein (NSP) specific antibodies to FMD virus (FMDV) by Ceditest FMDV-NSP Enzyme-lined Immunosorbent Assay (ELISA), and were confirmed by Liquid Phase Blocking ELISA (LPB ELISA). Results: Overall seroprevalence was 42.4%(n=184/434) by Ceditest-NSP ELISA, and 46.8%(n=203/434) by LPB ELISA against FMDV serotype O. The presence of antibodies against FMDV serotype O was higher (P<0.01) as compared to those of serotype A and Asia-1. The seroprevalence in Meikhtila (49.77%) was higher (P<0.01) than that of Pyawbwe (35.2%). The seropositivity in sheep and goats that were in-contact (77.19%) with infected cattle and pigs was higher (P<0.01) as compared to those in-contact with non-infected animals (37.14%). Similarly, the seropositivity in sheep and goats from high animal trade areas (49.4%) was higher (P<0.05) than that of those from low animal trade areas (37.97%). Conclusion: Rearing of sheep and goats in-contact with FMDV-infected cattle and pigs, and high animal trading areas are the major associated risk factors for FMDV infection for sheep and goats in the study areas in Myanmar. [ABSTRACT FROM AUTHOR]

This article details a systemic analysis of the controls in place and possible interventions available to further reduce the risk of a foot and mouth disease (FMD) outbreak in the United Kingdom. Using a research-based network analysis tool, we identify vulnerabilities within the multibarrier control system and their corresponding critical control points (CCPs). CCPs represent opportunities for active intervention that produce the greatest improvement to United Kingdom's resilience to future FMD outbreaks. Using an adapted 'features, events, and processes' (FEPs) methodology and network analysis, our results suggest that movements of animals and goods associated with legal activities significantly influence the system's behavior due to their higher frequency and ability to combine and create scenarios of exposure similar in origin to the U.K. FMD outbreaks of 1967/8 and 2001. The systemic risk assessment highlights areas outside of disease control that are relevant to disease spread. Further, it proves to be a powerful tool for demonstrating the need for implementing disease controls that have not previously been part of the system. [ABSTRACT FROM AUTHOR]

In recent years, foot-and-mouth disease virus (FMDV) serotype O, topotype Middle East-South Asia (ME-SA), lineage Ind-2001d has spread from the Indian subcontinent to the Middle East, North Africa, and Southeast Asia. In the current report, we describe the first detection of this lineage in Vietnam in May, 2015 in Đắk Nông province. Three subsequent outbreaks caused by genetically related viruses occurred between May–October, 2015 after which the virus was not detected in clinical outbreaks for at least 15 subsequent months. The observed outbreaks affected (in chronological order): cattle in Đắk Nông province, pigs in Đắk Lắk province and Đắk Nông province, and cattle in Ninh Thuận province. The clinical syndromes associated with these outbreaks were consistent with typical FMD in the affected species. Overall attack rate on affected premises was 0.85 in pigs and 0.93 in cattle over the course of the outbreak. Amongst 378 pigs at risk on affected premises, 85 pigs died during the outbreaks; there were no deaths among cattle. The manner in which FMDV/O/ME-SA/Ind-2001d was introduced into Vietnam remains undetermined; however, movement of live cattle is the suspected route. This incursion has substantial implications for epidemiology and control of FMD in Southeast Asia. [ABSTRACT FROM AUTHOR]

Availability of a rapid and sensitive diagnostic technique is key in successful prevention and control of infectious diseases like foot-and-mouth disease (FMD). Existing conventional diagnostic tests for FMD are laborious and time-consuming with low sensitivity and specificity. Molecular-based techniques are costly and difficult, involving refined apparatus like a thermal cycler. In the present study, the technique of real-time loop-mediated isothermal amplification (RT-LAMP) was standardized for diagnosing FMDV and its serotypes, evaluated using field samples, and compared with the existing real-time PCR in Pakistan. RT-LAMP amplified the target 3D gene using specific primers at 65 °C for 60 min and the VP1 gene using serotype specific primers at 63 °C for 60 min. A total of 38 samples out of 50 were positive by RT-LAMP and identified serotypes were A (n = 15), O (n = 15), and Asia-1 (n = 8). The efficiency of RT-LAMP in this study was highest for serotype Asia-1 (80.9%) followed by serotype A (73.4%) and O (62.35). [ABSTRACT FROM AUTHOR]

The present sero-epidemiological study was conducted in Gujarat plains and hills agro-climatic region of India during 2013-2014 to detect infection-specific antibodies against foot-and-mouth disease virus (FMDV) exposure in apparently healthy, unvaccinated, traditionally managed sheep and goat population across 26 districts of Gujarat state. Serum samples from 1,295 small ruminants (381 sheep and 914 goats) were collected and subjected to 3AB3 nonstructural protein (NSP) ELISA for detection of FMDV NSP antibodies (Abs); where 111 of 381 (29.13%) sheep and 201 of 914 (21.99%) goats were positive suggesting their previous exposure to FMDV. Although the animals were not vaccinated, representative numbers of serum samples from both species were also tested in liquid phase blocking ELISA to cross-check the protective antibody titre against all three serotypes in the trivalent vaccine that revealed null or zero 'herd immunity'. The results illustrate the circulation of FMDV in sheep and goats in the particular agro-climatic region of the country and vaccination in these animals needs to be followed to build up desired level of herd immunity against FMD. [ABSTRACT FROM AUTHOR]

Enterovirus 71 (EV71) is the major causative agent of hand-foot-andmouth disease in young children and can cause severe cerebral and pulmonary complications and even fatality. This study aimed at elucidating whether and how EV71 infection is regulated by a cellular microRNA, miR-127-5p. We found that miR-127-5p can downregulate the expression of SCARB2, a main receptor of EV71, by targeting two potential sites in its 3' UTR region and inhibit EV71 infection. Meanwhile, miR-127-5p expression was upregulated during EV71 infection. Notably, transfecting cells with miR-127-5p mimics led to a significant decrease in viral replication, while inhibition of endogenous miR-127-5p facilitated viral replication. Furthermore, our evidence showed that miR-127-5p did not affect postentry viral replication. Taken together, these results indicated that miR-127-5p inhibited EV71 replication by targeting the SCARB2 mRNA. [ABSTRACT FROM AUTHOR]

Germany has been free from foot-and-mouth disease virus (FMDV) without vaccination since 1992, but diagnostic capability at regional laboratories is maintained for FMDV exclusion in suspect cases and as surge capacity for outbreak preparedness. A proficiency test was initiated in 2015 to evaluate the diagnostic performance of 20 regional veterinary laboratories. A panel of two identical samples of FMDV genome for real-time reverse transcription polymerase chain reaction (RT-PCR), four lyophilized bovine sera for antibody detection and eight samples of inactivated vaccine antigen for analysis with a lateral-flow device (LFD) were tested with the systems routinely used at the participating institutions. With only one exception, all laboratories reliably detected viral RNA with two real-time RT-PCR assays down to a dilution of 10(−4) of the original material. The LFD pen-side test was evaluated at 8 of 20 laboratories, and FMDV antigen was detected by all participants down to a dilution of 1:81. Serological diagnosis was also very consistent at most participating institutions. Very few false-negative results were returned for the diluted positive sera, and testing of a large cohort of negative samples demonstrated a high specificity of over 99% for the two commercial ELISA kits used (PrioCHECK ® FMDV NS and ID Screen ® FMD NSP). In conclusion, the proficiency test demonstrated the reliable and robust FMDV diagnostic capability of the German regional veterinary diagnostic laboratories. Furthermore, the suitability of the commercially available ELISA systems for the detection of FMDV-specific antibodies was confirmed. Proficiency tests are an important quality assurance measure for transboundary diseases like FMDV where diagnostic capacity in free countries has to be available at the highest possible standard. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate a number of foot-and-mouth disease ( FMD) test methods for use in red deer. Ten animals were intranasally inoculated with the FMD virus ( FMDV) O UKG 11/2001, monitored for clinical signs, and samples taken regularly (blood, serum, oral swabs, nasal swabs, probang samples and lesion swabs, if present) over a 4-week period. Only one animal, deer 1103, developed clinical signs (lesions under the tongue and at the coronary band of the right hind hoof). It tested positive by 3 D and IRES real-time reverse transcription polymerase chain reaction (r RT- PCR) in various swabs, lesion materials and serum. In a non-structural protein ( NSP) in-house ELISA ( NSP- ELISA- IH), one commercial ELISA ( NSP- ELISA- PR) and a commercial antibody NSP pen side test, only deer 1103 showed positive results from day post-inoculation (dpi) 14 onwards. Two other NSP- ELISAs detected anti- NSP serum antibodies with lower sensitivity. It also showed rising antibody levels in the virus neutralization test ( VNT), the in-house SPO- ELISA- IH and the commercial SPO- ELISA- PR at dpi 9, and in another two commercial SPO- ELISAs at dpi 12 ( SPO- ELISA- IV) and dpi 19 ( SPO- ELISA- IZ), respectively. Six of the red deer that had been r RT- PCR and antibody negative were re-inoculated intramuscularly with the same O-serotype FMDV at dpi 14. None of these animals became r RT- PCR or NSP- ELISA positive, but all six animals became positive in the VNT, the in-house SPO- ELISA- IH and the commercial SPO- ELISA- PR. Two other commercial SPO- ELISAs were less sensitive or failed to detect animals as positive. The rRT- PCRs and the four most sensitive commercial ELISAs that had been used for the experimentally inoculated deer were further evaluated for diagnostic specificity ( DSP) using 950 serum samples and 200 nasal swabs from non-infected animals. DSPs were 100% for the rRT- PCRs and between 99.8 and 100% for the ELISAs. [ABSTRACT FROM AUTHOR]

Enteroviruses (EVs) are the etiological agents involved in most cases of hand, foot and mouth disease (HFMD) and herpangina (HA). Information on the epidemiology profiles of EVs in China is very limited, as the present surveillance system of China focuses on CAV16 and EV71, and no published data are available in Dongguan. The aim of this study is to determine the prevalence of EVs among patients with HFMD and HA in Dongguan, China, during 2015. A total of 271 clinical stool specimens that were clinically determined to be positive for enteroviruses were genotyped by semi-nested polymerase chain reaction (PCR) for the VP1 genes of EVs. The results showed that a total of 14 enterovirus genotypes were identified among HFMD and HA patients in this study. CVA6 was the most common genotype for HFMD, and CVA2 accounted for the majority of HA cases in this study. Sequence and phylogenetic analysis showed that all of the CVA6 and CVA2 strains identified in our study displayed a close genetic relationship to strains identified in other cities in China. This study also demonstrates that there are associations between particular causative enterovirus genotypes and some clinical symptoms, which may provide useful information for improving case prevention, diagnosis and treatment of HFMD and HA. [ABSTRACT FROM AUTHOR]

Background: Foot-and-mouth disease virus (FMDV) is one of the highest risk factors that affects the animal industry of the country. The virus causes production loss and high ratio mortality in young cloven-hoofed animals in Vietnam. The VP1 coding gene of 80 FMDV samples (66 samples of the serotype O and 14 samples of the serotype A) collected from endemic outbreaks during 2006-2014 were analyzed to investigate their phylogeny and genetic relationship with other available FMDVs globally. Results: Phylogenetic analysis indicated that the serotype O strains were clustered into two distinct viral topotypes (the SEA and ME-SA), while the serotype A strains were all clustered into the genotype IX. Among the study strains, the amino acid sequence identities were shared at a level of 90.1-100, 92.9-100, and 92.8-100% for the topotypes SEA, ME-SA, and genotype IX, respectively. Substitutions leading to changes in the amino acid sequence, which are critical for the VP1 antigenic sites were also identified. Our results showed that the studied strains are most closely related to the recent FMDV isolates from Southeast Asian countries (Myanmar, Thailand, Cambodia, Malaysia, and Laos), but are distinct from the earlier FMDV isolates within the genotypes. Conclusions: This study provides important evidence of recent movement of FMDVs serotype O and A into Vietnam within the last decade and their genetic accumulation to be closely related to strains causing FMD in surrounding countries. [ABSTRACT FROM AUTHOR]

Small ruminants (sheep and goats) are susceptible to foot-and-mouth disease (FMD), while studies with due emphasis on their role in the disease epidemiology have been meagre. The present study summarizes the results of clinico-molecular investigation of FMD in sheep and goats across several states of India during 2008–2014, where a total of 51 clinical epithelial tissue samples (vesicle/tongue/gum/foot epithelium) from sheep and 78 from goats were found positive for FMD virus (FMDV) serotype O in serotyping ELISA and multiplex reverse transcription-PCR. The VP1 region-based phylogenetic analysis demonstrated the involvement of O/ME-SA/Ind2001 lineage of serotype O virus in the outbreaks. The field viruses recovered from both small and large ruminant population during the same time period showed a close genetic relationship suggesting frequent inter-species transmission of virus. Since the disease often remains clinically camouflaged in small ruminants, the animals silently infected with FMD may pose potential threats to the in-contact livestock. Regular vaccination combined with surveillance and monitoring of protective antibody status in these species is therefore crucial to the effective control of FMD in the country. [ABSTRACT FROM AUTHOR]

Rapid, reliable and accurate diagnostic methods provide essential support to programmes that monitor and control foot-and-mouth disease (FMD). While pan-specific molecular tests for FMD virus (FMDV) detection are well established and widely used in endemic and FMD-free countries, current serotyping methods mainly rely either on antigen detection ELISAs or nucleotide sequencing approaches. This report describes the development of a panel of serotype-specific real-time RT-PCR assays (rRT-PCR) tailored to detect FMDV lineages currently circulating in East Africa. These assays target sequences within the VP1-coding region that share high intra-lineage identity, but do not cross-react with FMD viruses from other serotypes that circulate in the region. These serotype-specific assays operate with the same thermal profile as the pan-diagnostic tests making it possible to run them in parallel to produce C T values comparable to the pan-diagnostic test detecting the 3D-coding region. These assays were evaluated alongside the established pan-specific molecular test using field samples and virus isolates collected from Tanzania, Kenya and Ethiopia that had been previously characterised by nucleotide sequencing. Samples (n = 71) representing serotype A (topotype AFRICA, lineage G-I), serotype O (topotypes EA-2 and EA-4), serotype SAT 1 (topotype I (NWZ)) and serotype SAT2 (topotype IV) were correctly identified with these rRT-PCR assays. Furthermore, FMDV RNA from samples that did not contain infectious virus could still be serotyped using these assays. These serotype-specific real-time RT-PCR assays can detect and characterise FMDVs currently circulating in East Africa and hence improve disease control in this region. [ABSTRACT FROM AUTHOR]

Two duplex one-step TaqMan-based RT-PCR protocols for detection of foot-and-mouth disease virus (FMDV) were established and validated. Each RT-PCR test consists of a ready-to-use master mix for simultaneous detection of the well established 3D or IRES FMDV targets and incorporates the host β-actin mRNA as an internal control target, in a single-tube assay. The two real-time RT-PCR 3D/β-actin and IRES/β-actin tests are highly sensitive and able to detect up to 7 TCID 50 /ml of FMDV and 10 copies/1 μl of viral RNA. In field epithelium samples, the diagnostic sensitivity was 100% (95% CI; 91–100%) for the 3D/β-actin test and 97% (95% CI; 87–100%) for the IRES/β-actin test. The diagnostic specificity was 100% (95% CI; 95–100%) for both RT-PCRs. In addition, the two protocols proved to be robust, showing inter-assay coefficients of variation ranging from 1.94% to 6.73% for the IRES target and from 2.33% to 5.42% for the 3D target for different RNA extractions and different RT-PCR conditions. The internally controlled one-step real-time RT-PCR protocols described in this study provide a rapid, effective and reliable method for the detection of FMDV and thus may improve the routine diagnosis for foot-and-mouth disease. [ABSTRACT FROM AUTHOR]

Foot-and-mouth disease (FMD) is endemic in many regions of the world and is one of the most prevalent epizootic animal diseases. FMD affects livestock, such as cattle, sheep, goats and pigs, and causes enormous economic losses due to reduced productivity and trade restrictions. Preparedness and early diagnosis are essential for effective control of FMD. Many diagnostic assays are dependent on raising high-affinity, anti-FMD virus (FMDV) serotype-specific antibodies in small animals (rabbits and guinea pigs) that give broad virus coverage. Here we show that soluble, truncated forms of bovine αvβ6 bind FMDV in an authentic RGD and divalent cation dependent interaction and can be used as the trapping reagent in a FMDV sandwich ELISA. In addition, inclusion of FLAG or His tags facilitates simple purification without the loss of virus binding. We also provide evidence that when combined with a guinea pig polyclonal serum, or serotype-specific monoclonal antibodies, the integrin can be used to detect viruses representative of all FMDV serotypes. We also show that recombinant FMDV empty capsids, with stabilising disulphide bonds, can serve as an antigen in the ELISA and can therefore replace inactivated virus antigen as a positive control for the assay. Our results demonstrate the potential use of bovine αvβ6 and FMDV empty capsids in FMD diagnostic assays. [ABSTRACT FROM AUTHOR]

Foot-and-mouth disease (FMD) is a highly contagious epidemic disease of transboundary importance. Inadequate storage and shipment of suspected clinical samples can compromise the ability to detect and characterise FMD virus (FMDV) in endemic countries, thereby, leading to the loss of valuable virological and epidemiological data. This study, investigates the potential of using FTA ® cards for dry transportation of clinical samples and subsequent recovery of infectious FMDV by chemical transfection of FTA ® card fixed RNA as an alternative to the conventional cell culture based virus isolation method. A higher proportion of infectious FMDV was rescued from clinical samples (cell culture isolates, tongue epithelial suspension and impression smears) by the FTA ® card fixed RNA transfection method (76%) compared to the conventional cell culture based virus isolation (56%), suggesting a better performance of the current RNA transfection procedure. Furthermore, it was possible to rescue live virus by the transfection of RNA extracted from FTA ® card impregnated with clinical samples that had been stored at varying temperature (4–37 °C) up to a period of six weeks. The VP1 sequence data and antigenic relationships with the vaccine strains, between viruses rescued by FTA ® card fixed RNA transfection and conventional cell culture, were comparable. Therefore, these results support the use of the FTA ® card for the economic, dry, non-hazardous transport of FMD suspected clinical samples from the site of collection to national/international reference laboratories. [ABSTRACT FROM AUTHOR]

Periodic fever, aphthous stomatitis, pharyngitis, and adenitis (PFAPA) syndrome, the most common periodic disorder of childhood, presents with the cardinal symptoms of periodic fever, aphthous stomatitis, pharyngitis, and adenitis typically before age 5. This review presents the recent literature on PFAPA and summarizes key findings in the pathogenesis, evaluation, and treatment of the disease. Theories surrounding the pathogenesis of PFAPA include a faulty innate immunologic response in conjunction with dysregulated T-cell activation. A potential genetic link is also under consideration. Mediterranean fever (MEFV) gene variants have been implicated and appear to modify disease severity. In individuals with the heterozygous variant, PFAPA episodes are milder and shorter in duration. Diagnostic criteria include the traditional clinical signs, in addition to the following biomarkers: elevated C-reactive protein in the absence of elevated procalcitonin, vitamin D, CD64, mean corpuscular volume, and other nonspecific inflammatory mediators in the absence of an infectious explanation for fever. Treatment of PFAPA includes tonsillectomy, a single dose of corticosteroids, and, most recently, interleukin 1 blockers such as anakinra, rilonacept, and canakinumab. Tonsillectomy remains the only permanent treatment modality. [ABSTRACT FROM AUTHOR]

Foot and mouth disease (FMD) is a highly contagious viral disease of all cloven footed domestic and wild animals. This work was planned to study the different markers for diagnosis of FMDV serotype SAT 2 in adult buffaloes and calves including clinical, hematological and biochemical examinations. Forty adult buffaloes 3 to 5 years old of age were divided into two groups of twenty buffaloes; the first group was obvious clinically healthy and the second was naturally infected with FMD virus (FMDV). In addition, twenty suckling calves 10 to 50 days old of age were divided into two groups of ten calves the clinically apparent healthy group and the naturally infected with the FMDV group. The recorded clinical signs were fever, salivation, loss of appetite, depression, lameness, vesicles, erosions and ulcers in the mucosa of the mouth, tongue, lips, gums, palate and between the claws in infected adult. Development of macrocytic normochromic anemia. Leucopenia, lymphopenia and monocytopenia were recorded in infected adult buffaloes and calves. Myocardial injury proved by presence of degenerated myocardial fibers and lymphocyte cell infiltration with a significant increase in cardiac markers as cardiac torponin I (cTnI), creatine phosphokinase (CPK) and lactate dehyrogenase (LDH) in addition to a significant hyperkalemia, hypocalcaemia and hypomagnesaemia in buffalo calves. Moreover, electrophoresis showed hyoproteinemia, hypoalbuminemia and hypoglobulinemia in the infected animals. It could be concluded that the elevation of cardiac markers emphasized that FMD is more severe in young calves than adult buffaloes. Therefore, it is recommended to evaluate the prognosis of FMD infection in calves by these markers. [ABSTRACT FROM AUTHOR]

This study assessed knowledge gaps in foot-and-mouth disease ( FMD) research in the field of diagnostics. The study took the form of a literature review (2011-15) combined with research updates collected in 2014 from 33 institutes from around the world. Findings were used to identify priority areas for future FMD research. Molecular and genetic technologies, including sequencing, are developing at an increasing rate both in terms of capability and affordability. These advances potentiate progress in many other fields of research, from vaccine development to epidemiology. The development of RT- LAMP represents an important breakthrough allowing greater use and access to molecular diagnostics. It is now possible to determine virus serotype using PCR, although only for certain virus pools, continued progress is needed to cover the global spectrum of FMD viruses. Progress has also been made in the development of pen-side rapid diagnostics, some with the ability to determine serotype. However, further advances in pen-side serotype or strain determination would benefit both FMD-free countries and endemic countries with limited access to well-resourced laboratories. Novel sampling methods that show promise include air sampling and baited ropes, the latter may aid sampling in wildlife and swine. Studies of infrared thermography for the early detection of FMD have not been encouraging, although investigations are ongoing. Multiplex tests have been developed that are able to simultaneously screen for multiple pathogens with similar clinical signs. Crucial for assessing FMDV freedom, tests exist to detect animals that have been infected with FMDV regardless of vaccination status; however, limitations exist, particularly when testing previously vaccinated animals. Novel vaccines are being developed with complementary DIVA tests for this purpose. Research is also needed to improve the current imprecise approaches to FMD vaccine matching. The development of simple, affordable tests increases access to FMD diagnostics, greatly benefiting regions with limited laboratory capacity. [ABSTRACT FROM AUTHOR]

The Global Foot-and-mouth disease ( FMD) Research Alliance periodically reviews the state of FMD research to assess progress and to identify new priorities. In this supplement we provide an update of global FMD research, comprising (i) this overview paper, which includes background information with key findings, and papers covering (ii) epidemiology, wildlife and economics, (iii) vaccines, (iv) diagnostics, (v) biotherapeutics and disinfectants, (vi) immunology and (vii) pathogenesis and molecular biology. FMD research publications were reviewed (2011-2015) and activity updates were obtained from 33 FMD research institutes from around the world. Although a continual threat, FMD has been effectively controlled in much of the world using existing tools. However, control remains a challenge in most developing countries, where little has been done to understand the ongoing burden of FMD. More research is needed to support control in endemically infected countries, particularly robust field studies. Traditional FMD vaccines have several limitations including short duration and spectrum of protection, cold chain requirements, and the costs and biosecurity risks associated with vaccine production. Significant progress has been made in the development of novel vaccine candidates, particularly in the use of recombinant vaccines and virus-like particles as an alternative to traditional inactivated whole virus vaccines. Continued investment is needed to turn these developments into improved vaccines produced at scale. Increased knowledge of cellular and mucosal immunity would benefit vaccine development, as would further advances in our ability to enhance vaccine capsid stability. Developments in molecular biology and phylogenetics underlie many of the recent advances in FMD research, including improved vaccines and diagnostics, and improved understanding of FMD epidemiology. Tools for genetic analyses continue to become both more powerful and more affordable enabling them to be used to address an ever-expanding range of questions. This rapidly advancing field potentiates many areas of FMD research and should be prioritized. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate strategies to enhance the early detection of foot and mouth disease incursions in Australia. Two strategies were considered. First, improving the performance of the current passive surveillance system. Second, supplementing the current passive system with active surveillance strategies based on testing animals at saleyards or through bulk milk testing of dairy herds. Simulation modelling estimated the impact of producer education and awareness by either increasing the daily probability that a farmer will report the presence of diseased animals or by reducing the proportion of the herd showing clinical signs required to trigger a disease report. Both increasing the probability of reporting and reducing the proportion of animals showing clinical signs resulted in incremental decreases in the time to detection, the size and the duration of the outbreak. A gold standard system in which all producers reported the presence of disease once 10% of the herd showed clinical signs reduced the median time to detection of the outbreak from 20 to 15 days, the duration of the subsequent outbreak from 53 to 42 days and the number of infected farms from 46 to 32. Bulk milk testing reduced the median time to detection by two days and the number of infected farms by six but had no impact on the duration of the outbreak. Screening of animals at saleyards provided no improvement over the current passive surveillance system alone while having significant resource issues. It is concluded that the most effective way to achieve early detection of incursions of foot and mouth disease into Victoria, Australia is to invest in improving producer reporting. [ABSTRACT FROM AUTHOR]

In foot-and-mouth disease (FMD)-endemic parts of the globe, control is mainly implemented by preventive vaccination with an inactivated purified vaccine. ELISAs detecting antibodies to the viral nonstructural proteins (NSP) distinguish FMD virus (FMDV)-infected animals in the vaccinated population (DIVA). However, residual NSPs present in the vaccines are suspected to be a cause of occasional false positive results, and therefore, an epitope-deleted negative marker vaccine strategy is considered a more logical option. In this study, employing a serotype Asia 1 FMDV infectious cDNA clone, it is demonstrated that while large deletions differing in size and location in the carboxy-terminal half of 3A downstream of the putative hydrophobic membrane-binding domain (deletion of residues 86-110, 101-149, 81-149 and 81-153) are tolerated by the virus without affecting its infectivity in cultured cell lines, deletions in the amino-terminal half (residues 5-54, 21-50, 21-80, 55-80 and 5-149) containing the dimerization and the transmembrane domains are deleterious to its multiplication. Most importantly, the virus could dispense with the entire carboxy-terminal half of 3A (residues 81-153) including the residues involved in the formation of the 3A-3B cleavage junction. The rescue of a replication-competent FMDV variant carrying the largest deletion ever in 3A (residues 81-153) and the fact that the deleted region contains a series of linear B-cell epitopes inspired us to devise an indirect ELISA based on a recombinant 3A carboxy-terminal fragment and to evaluate its potential to serve as a companion diagnostic assay for differential serosurveillance if the 3A-truncated virus is used as a marker vaccine. [ABSTRACT FROM AUTHOR]

Over a decade ago, foot-and-mouth disease (FMD) re-emerged in Southern Africa specifically in beef exporting countries that had successfully maintained disease-free areas in the past. FMD virus (FMDV) serotype SAT2 has been responsible for a majority of these outbreaks. Epidemiological studies have revealed the importance of the African buffalo as the major wildlife FMD reservoir in the region. We used phylogeographic analysis to study dynamics of FMD transmission between buffalo and domestic cattle at the interface of the major wildlife protected areas in the region currently encompassing two largest Transfrontier conservation areas: Kavango-Zambezi (KAZA) and Great Limpopo (GL). Results of this study showed restricted local occurrence of each FMDV SAT2 topotypes I, II, and III, with occasional virus migration from KAZA to GL. Origins of outbreaks in livestock are frequently attributed to wild buffalo, but our results suggest that transmission from cattle to buffalo also occurs. We used coalescent Bayesian skyline analysis to study the genetic variation of the virus in cattle and buffalo, and discussed the association of these genetic changes in the virus and relevant epidemiological events that occurred in this area. Our results show that the genetic diversity of FMDV SAT2 has decreased in buffalo and cattle population during the last decade. This study contributes to understand the major dynamics of transmission and genetic variation of FMDV SAT2 in Southern Africa, which will could ultimately help in designing efficient strategies for the control of FMD at a local and regional level. [ABSTRACT FROM AUTHOR]

A systematic study was performed to investigate the potential of pigs to establish and maintain persistent foot-and-mouth disease virus ( FMDV) infection. Infectious virus could not be recovered from sera, oral, nasal or oropharyngeal fluids obtained after resolution of clinical infection with any of five FMDV strains within serotypes A, O and Asia-1. Furthermore, there was no isolation of live virus from tissue samples harvested at 28-100 days post-infection from convalescent pigs recovered from clinical or subclinical FMD. Despite lack of detection of infectious FMDV, there was a high prevalence of FMDV RNA detection in lymph nodes draining lesion sites harvested at 35 days post-infection, with the most frequent detection recorded in popliteal lymph nodes (positive detection in 88% of samples obtained from non-vaccinated pigs). Likewise, at 35 dpi, FMDV capsid antigen was localized within follicles of draining lymph nodes, but without concurrent detection of FMDV non-structural protein. There was a marked decline in the detection of FMDV RNA and antigen in tissue samples by 60 dpi, and no antigen or viral RNA could be detected in samples obtained at 100 dpi. The data presented herein provide the most extensive investigation of FMDV persistence in pigs. The overall conclusion is that domestic pigs are unlikely to be competent long-term carriers of infectious FMDV; however, transient persistence of FMDV protein and RNA in lymphoid tissues is common following clinical or subclinical infection. [ABSTRACT FROM AUTHOR]

Foot-and-mouth disease (FMD) is caused by the FMD virus (FMDV) and results in severe economic losses in livestock farming. For rapid FMD diagnostic and therapeutic purposes, an effective antibody against FMDV is needed. Here, we developed a high-affinity antibody against FMDV by FACS-based high throughput screening of a random library. With the FITC-conjugated VP1 epitope of FMDV and high-speed FACS sorting, we screened the synthetic antibody (scFv) library in which antibody variants are displayed in the periplasm of Escherichia coli . After three rounds of sorting, we isolated one antibody fragment (#138-scFv) against the VP1 epitope of FMDV. Next, to improve its affinity, a mutation library of #138-scFV was constructed by error-prone PCR and screened by FACS. After three rounds of sorting, we isolated one antibody (AM-32 scFv), which has a higher binding affinity ( K D = 42.7 nM) than that of the original #138-scFv. We also confirmed that it specifically binds to whole inactivated FMDV. [ABSTRACT FROM AUTHOR]

Humans living in and around forest areas are at increased risk of zoonotic disease transmission. The transhumant Van Gujjars of North India are one such population, but there is an absence of health data, including evidence of zoonotic diseases, in this community. Pastoral communities can have a wide breadth of knowledge of livestock diseases, but not necessarily of human diseases. This study investigated the perceptions that the Van Gujjars have specifically of zoonotic diseases, using participatory epidemiological methods, including semi-structured interviews, ranking, proportional piling, transect walks and direct observation, triangulated by informal interviews with local veterinarians. The community did not have a wide appreciation of zoonotic diseases, apart from rabies and potentially zoonotic skin diseases. In contrast, animal diseases were of much greater concern to the community; the locally-named surra (trypanosomiasis), ajar, khuriya (foot-and-mouth disease) , dakhutra, gheru, taku, and ‘ blood in urine ’ (possibly babesiosis), being of most concern. A participatory epidemiological approach was found to be an effective method of data collection and analysis; and the findings suggest that access to health services, particularly veterinary health services, should be improved for Van Gujjars. [ABSTRACT FROM AUTHOR]

The time delay to detection of an outbreak of an emergency animal disease directly affects the size of the outbreak at detection and the likelihood that the disease can be eradicated. This time delay is a direct function of the efficacy of the surveillance system in the country involved. Australia has recently completed a comprehensive review of its general surveillance system examining regional variation in both the behaviour of modelled outbreaks of foot and mouth disease and the likelihood that each outbreak will be detected and reported to government veterinary services. The size of the outbreak and the time delay from introduction to the point where 95% confidence of detection was reached showed significant (p < 0.05) regional variation with the more remote northern areas experiencing smaller outbreaks that are less likely to spread and less likely to be reported to government services than outbreaks in the more developed southern areas of Australia. Outbreaks in the more densely populated areas may take up to 43 days until a 95% confidence of detection is achieved and at that time, the outbreak may involve up to 53 farms. [ABSTRACT FROM AUTHOR]

Hand, foot, and mouth disease (HFMD) is an infectious disease caused by human enterovirus 71 (EV71), coxsackievirus A16 (CVA16) and other enteroviruses. It is of interest that other enteroviruses associated with HFMD in Jinan have been rarely reported. The aim of the present study is to detect and characterize the circulating serotypes of non-EV71 and non-CVA16 enteroviruses associated with HFMD in Jinan city, Shandong province, China. A total of 400 specimens were collected from clinically diagnosed HFMD cases in Jinan from January 2009 to June 2013. All specimens were infected with non-EV71 and non-CVA16 enteroviruses previously confirmed by RT-PCR or real-time PCR according to the protocols at that time. The GeXP-based multiplex RT-PCR assay (GeXP assay) was performed to investigate the pathogen spectrum of 15 enteroviruses (coxsackieviruses A4, A5, A6, A9, A10, A16; coxsackieviruses B1, B3, B5; Echoviruses 6, 7, 11, 13, 19 and EV71) infections associated with HMFD. For GeXP assay negative samples, reverse transcription nested PCR (nested RT-PCR) based on the 5’ -untranslated region (5’- UTR) sequence and phylogenetic analysis were conducted to further explore the etiology of multiple enteroviruses. The results showed that a total of twenty serotypes of enteroviruses (including EV71 and CVA16) were identified by GeXP assay and nested RT-PCR. The most circulating twelve serotypes of enteroviruses with HFMD in Jinan from 2009 to June 2013 were EV71, CVA16, CVA10, CVA6, CVA12, CVA2, Echo3, CVA4, CVA9, CVB1, CVB3 and Echo6. CVA10 and CVA6 were the most prevalent pathogens other than EV71 and CVA16 in Jinan and their most prevalent seasons were spring and summer, and a slight increase was observed in autumn and early winter. It should be noted that mixed-infections were identified by GeXP assay and the phylogenetic tree clearly discriminated the multiple pathogens associated with HFMD. Our results thus demonstrate that there was a clear lack of a reliable testing method for EV71 and CVA16 and multiple non-EV71 and non-CVA16 enteroviruses associated with HFMD were present in Jinan. The GeXP assay combined with nested RT-PCR based on 5’-UTR region could meet the need for the national surveillance of multiple enteroviruses or the investigation of epidemic outbreaks triggered by enteroviruses in the future. [ABSTRACT FROM AUTHOR]

Foot-and-mouth disease (FMD) is a highly contagious disease of cloven-hoofed animals. FMD virus (FMDV) shows a strong tropism for epithelial cells, and FMD is characterised by cell lysis and the development of vesicular lesions in certain epithelial tissues (for example, the tongue). By contrast, other epithelial tissues do not develop lesions, despite being sites of viral replication (for example, the dorsal soft palate). The reasons for this difference are poorly understood, but hypotheses are difficult to test experimentally. In order to identify the factors which drive cell lysis, and consequently determine the development of lesions, we developed a partial differential equation model of FMDV infection in bovine epithelial tissues and used it to explore a range of hypotheses about epithelium structure which could be driving differences in lytic behaviour observed in different tissues. Our results demonstrate that, based on current parameter estimates, epithelial tissue thickness and cell layer structure are unlikely to be determinants of FMDV-induced cell lysis. However, differences in receptor distribution or viral replication amongst cell layers could influence the development of lesions, but only if viral replication rates are much lower than current estimates. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]