The Complete Genomic Sequence of a Foot-and-Mouth Disease Virus Isolated from the Swine

The complete genomic sequence of foot-and-mouth disease virus (FMDV) Chinese strain OH/CHA/99 was determined. The 8040 nt sequence and the deduced amino acid sequence werecompared with FMDV sequences published. The results showed that OH/CHA/99 shared highersequence homology with OTYTW/97, indicating their close genetic relationship. However,the strain had lower sequence identity with O1/Kaufbeuren/66 strain. Besides, largedeletions in 3A coding region were observed in OH/CHA/99. It was shown that the poly (A)tail of OH/CHA/99 had 56 As at least.

Comparison of Three ELISA Kits for the Differentiation of Foot-and-mouth Disease Virus-infected from Vaccinated Animals

研究被执行验证工具包为 FMDV 的区别在中国开发了的 3 FMDV 3ABC-I-ELISA 感染并且种牛痘的动物。从天真、种牛痘的牛以及从被感染了的牛的 sera 的集合被商业诊断工具包对 FMDV 的 nonstructural 蛋白质(NSP ) 为抗体测试， Ceditest 吗？FMDV-NS (Ceditest？工具包) ， UBI？FMDV NONSTRUCTURAL 蛋白质 ELISA 方向插入(UBI？工具包) 并且一个 FMDV 3ABC-I-ELISA 工具包在 Lanzhou 发展了兽医研究院。三个工具包的测试参数(敏感和特性) 被决定，并且从 FMD 3ABC-I-ELISA 工具包获得的结果与从二个外国工具包获得了那相比。结果显示巧合在 FMDV 3ABC-I-ELISA 和 Ceditest 之间评价吗？工具包 98.05% 是，并且在 FMDV 3ABC-I-ELISA 和 UBI 之间的巧合率？工具包是 94.4% ；两 Ceditest 的敏感？并且 FMDV 3ABC-I-ELISA 工具包是 100% 。然而， UBI 的敏感？工具包仅仅是 81.8% 。与从天真或种牛痘的非感染的动物的 sera，所有测试的特性超过了 90% 。关键词 Foot-and-mouth 疾病病毒(FMDV )- 非结构的蛋白质 3ABC - ELISA CLC 数字 S852.65 基础项目：国家鈥 ? 73 鈥 ? 研究工程(G199901190， 2005CB23201 ) ；国际原子能机构(国际原子能机构)(10697/R2 )

Assessment of immunization procedures for foot-and-mouth disease in large-scale pig farms in China based on actual data and dynamics

Foot-and-mouth disease(FMD)is an acute,highly infectious and pathogenic animal disease.In recent years,with the rapid development of the swine breeding industry in China,pig farms have shown a trend of larger-scale development.Large-scale pig farms employ standardized management,a high level of automation,and a strict_system.However,these farms have a large trading volume,and increased transmission intensity of FMD is noted inside the farm.At present,the main control measure against FMD is pig vaccination.However,a standard for immunization procedures is not available,and currently adopted immunization procedures have not been effectively and systematically evaluated.Taking a typical large-scale pig farm in China as the research subject and considering the breeding pattern,piggery structure,age structure and immunization procedures,an individual-based state probability model is established to evaluate the effectiveness of the immune procedure.Based on numerical simulation,it is concluded that the optimal immunization program involves primary immunization at 40 days of age and secondary immunization at 80 days of age for commercial pigs.Breeding boars and breeding sows are immunized 4 times a year,and reserve pigs are immunized at 169 and 259 days of age.According to the theoretical analysis,the average control reproduction number of individuals under the optimal immunization procedure in the farm is 0.4927.In the absence of immunization,the average is 1.7498,indicating that the epidemic cannot be controlled without immunization procedures.

Effect of Attenuated Highly Pathogenic Pig Reproductive and Respiratory Syndrome(HP-PRRS) TJM-F92 Strain Vaccine on Immune Antibody Levels against Classical Swine Fever(CSF) and Foot-and-Mouth Disease(FMD)

Effects of attenuated highly pathogenic pig reproductive and respiratory syndrome（HP-PRRS）TJM-F92 strain vaccine on immune antibody level against classical swine fever（CSF）and foot-and-mouth disease（FMD）were studied from October 8 to November 12 in 2014,in order to optimize vaccination program of CSF,HP-PRRS and FMD and to provide scientific guidance for animal disease control and prevention work.The results showed that attenuated HP-PRRS（TJMF92 strain）vaccine had no significant effect on immune antibody level of hog cholera lapinized virus（HCLV,ST passage cell vaccine）attenuated vaccine and FMD-O inactivated vaccines（OZK/93 strain）,and single or combined use of three vaccines received good immunization effects.

Expression,Purification and Activity Detection of Structural Protein VP1 of Foot-and-Mouth Disease Virus Serotype A

The paper was to obtain the VP1 protein of FMDV serotype A with high activity. With recombinant plasmid pMD19A-T-vp1 as the tem- plate, vpl gene fragment amplified by PCR was connected into prokaryotic expression vector pET28a to construct recombinant plasmid pET-A-vpl. The E. coli BL21 （DE3） strain containing recombinant plasmid pET-A-vpl were induced by IPTG. SDS-PAGE showed that VP1 protein was ex- pressed in the form of inclusion body, and its molecular weight was about 29 ku. Based on the optimizing IPTG concentration and expression time, the largest expression of VP1 protein was induced by 0.3 mmol/L IPTG for 6 h at 37 ℃. Western-Blot analysis indicated that the expression of VP1 protein could be specifically recognized by positive serum of FMDV serotype A. ELISA test showed that VP1 inclusion body protein had high activity after purification by washing and renaturation by urea concentration gradient dialysis.

Molecular and Serological Epidemiology of Foot-and-Mouth Disease Virus in North Region of Cameroon

The serological prevalence of foot and mouth disease virus (FMDV) among the cattle population in the North region of Cameroon was determined using ELISA (Enzyme Linked Immunosorbent Assays) serological tests for structural as well as non-structural proteins. In these cattle, FMDV RNA was identified, amplified, sequenced and the sequences were used to construct a phylogenetic tree. A sedentary cattle population randomly selected from six veterinary centres in the North region was sampled twice, six months apart. High prevalence of FMDV antibody was recorded in the first (402/466 (85.84%)) and second (358/411 (86.90%)) sampling periods. There was no significant difference (P > 0.05) in prevalence of FMDV antibody between the two sampling periods. Goudali and Peulh breeds of cattle and animals of three to five years old were the most infected with FMDV and mostly in the months of May and August. A seroprevalence of 100% (n = 14) of FMDV against serotypes A and O was observed in sera from convalescent animals in the study area. FMDV antigen detection ELISA showed a prevalence of 18/37 (48.65%) for serotypes SAT1 (8.1%), SAT2 (35.1%), A (10.8%) and O (2.7%) among the clinically infected animals. There was no significant difference (P > 0.05) in prevalence of FMDV RNA between the sampling periods. A prevalence of FMDV RNA (17.5% (n = 120) and 16.7% (n = 240)) was observed among the sedentary animals that were sampled four to five months apart. FMDV RNA prevalence of 28/37 (75.6%) among clinically infected animals was also observed, thus confirming all the 12 outbreaks investigated. Sequence analysis of VP1 coding gene of the SAT2 serotype showed that it was homologous to the Libyan isolates (that caused epidemics in northern Africa in 2012) and also clustered with the serotypes isolated from both Nigeria and Sudan in 2007.

Geographic Information System to Monitor and Forecast the Influence of Foot-and-Mouth Disease of Poultry and Cattle

The paper introduces the origin,technical features,scale,application efficiency of geographic information system(GIS)to monitor the situation and forecast the spread of foot-and-mouth disease of poultry and cattle—a technological product of Vietnam Software Academy for Water.Application of the system will help the producers and managers to quickly capture the situation of raising poultry and cattle,vaccination results in the related territory,flu outbreaks,epidemic foot-and-mouth disease,and the spread of these two dangers as they arise,resulting in appropriate decisions and solutions in order to prevent,minimize the risk effectively.

Space-Time Analysis of Foot-and-Mouth Disease from 2005 to 2014 among Cattle Populations in Benin:A Retrospective Study

Many cases of foot-and-mouth disease (FMD) are reported every year in Benin. In order to elucidate the epidemiology of this disease, a space-time analysis was carried out in all the 77 municipalities of the country aiming to identify high risk areas as well as risk factors such as season and transhumance on the period of 2005 to 2014. Data were collected retrospectively from the Directory of Animal Production of Benin. The method of Kulldorff was used with the software SaTScanTM for the space-time analysis while a script was designed in the software R to generate new sizes with three different models of transhumance. From 2005 to 2014, 434 foci were recorded. Many outbreaks occurred in August, September and October. This period corresponds to the small rainy season in the South and the rainy season of the North. The municipality of Parakou was regarded as the source FMD outbreaks in Benin because it hosts one of the largest livestock markets in the country and many rivers. It was the municipality at the highest risk. The other municipalities at risk were Nikki, Pèrèrè and Kalaléas well as Karimama (hosting the national parkW), Kouandé and Toucountouna located nearby Pendjari’s national park. This study revealed that the space-time configuration is real and the main factors of persistence and dissemination of FMD virus were national parks, classified forests and the livestock market of Parakou all located in the North. The variation of the number of cattle due to their transhumance from the North to the South did not influence the zones at risk. Therefore, Northern Benin is probably at high risk of FMD.

Effect of amino acid mutation at position 127 in 3A of a rabbitattenuated foot-and-mouth disease virus serotype Asia1 on viral replication and infection

An amino acid mutation(R127→I) in the 3A non-structural protein of an FMDV serotype Asia1 rabbit-attenuated ZB strain was previously found after attenuation of the virus. To explore the effects of this mutation on viral replication and infection, the amino acid residue isoleucine(I) was changed to arginine(R) in the infectious cDNA clone of the rabbit-attenuated ZB strain by sitedirected mutagenesis, and the R127-mutated virus was rescued. BHK monolayer cells and suckling mice were inoculated with the R127-mutated virus to test its growth property and pathogenicity, respectively. The effects of the R127 mutation on viral replication and virulence were analyzed. The data showed that there was a slight difference in plaque morphology between the R127-mutated and wild-type viruses. The growth rate of the mutated virus was lower in BHK-21 cells and its virulence in suckling mice was also attenuated. This study indicates that the R127 mutation in 3A may play an important role in FMDV replication in vitro and in pathogenicity in suckling mice.

Adverse Effects of Inactivated Foot-and-Mouth Disease Vaccine—Possible Causes Analysis and Countermeasures

Foot-and-mouth disease (FMD) is an infectious and sometimes fatal viral disease that affects cloven-hoofed animals, and Chinese government adopts compulsory immunization measures for FMD. The adverse effects of FMD vaccine to pigs, cattle and goats have been reported increasingly frequent during the spring and autumn seasons when large numbers of farm livestock are vaccinated. The financial losses caused by vaccine adverse effects have been a serious concern for both farmers and primary prevention personnel. There are various causative factors reported to involve into adverse effect of FMD vaccine, including the inappropriate vaccine production, transportation and storage, livestock poor tolerance, and unqualified vaccinating manipulations. Symptomatic treatment and early drug prevention have a certain effect on the adverse effects. To analyze causes and propose countermeasures, in the current study possible reasons during the production and processing procedures of inactivated FMD vaccine were reviewed and corresponding countermeasures were recommended. The review may provide references for better use of vaccine to prevent FMD.

Review of Air Dispersion Modelling Approaches to Assess the Risk of Wind-Borne Spread of Foot-and-Mouth Disease Virus

Foot-and-mouth disease virus (FMDV) is one of the most economically serious veterinary pathogens due to its negative effects on livestock and its highly infectious nature via a variety of transmission paths through oral and inhalation routes. Measures to enhance outbreak management can be designed according to analytical results predicted by mathematical models for wind-borne dispersion, an important path of virus transmission. Accurate atmospheric dispersion models are useful tools for properly determining risk management plans, while inaccurate models may conversely lead to accidental loss in two possible ways. Overly strict measures, e.g., slaughter for too wide an area, can cause severe economic difficulties, including irreversible loss of business operations for a number of farms. On the contrary, inestimable loss potentially caused by lax controls is a persistent threat. In this paper, available modelling procedures for forecasting the spread of FMDV, which have been used since the 1970s, each having its advantages and limitations, are reviewed for the purpose of ensuring suitable application in various conditions of any future emergency cases.

Pancreatitis in hand-foot-and-mouth disease caused byenterovirus 71

Some viruses, including certain members of the enterovirus genus, have been reported to cause pancreatitis, especially Coxsackie virus. However, no case of human enterovirus 71(EV71) associated with pancreatitis has been reported so far. We here report a case of EV71-induced hand-foot-and-mouth disease(HFMD) presenting with pancreatitis in a 2-year-old girl. This is the first report of a patient with acute pancreatitis in HFMD caused by EV71. We treated the patient conservatively with nasogastric suction, intravenous fluid and antivirals. The patient's symptoms improved after 8 d, and recovered without complications. We conclude that EV71 can cause acute pancreatitis in HFMD, which should be considered in differential diagnosis, especially in cases of idiopathic pancreatitis.

Development of a Synthetic Peptide ELISA Assay for the Detection of Foot-and-Mouth Disease Virus Nonstructural Protein Antibodies

In order to develop an ELISA assay with synthetic peptides for the detection of antibody to the nonstructural proteins of foot-and-mouth disease virus, specific peptides were synthesized by a solid-phase method according to FMDV NSPs B-cell epitopes, and were conjugated to carrier protein BSA. An ELISA system was developed to detect FMDV NSPs antibody with the conjugated proteins as the coating antigen. The optimal coating concentration of the antigen was determined as 2.5 μg mL-1. The comparative study of this assay with UBI NSP ELISA kit and national commercial 3ABC ELISA kit in the detection of 199 serum samples showed that they were very coincident, and the identity rates were 96.48 and 97.48%, respectively. The development of ELISA using the synthetic peptides as coating antigen is specific, reproducible, stable, and easy, and can be used to differentiate FMDV infected pigs from immunized pigs.

Comparison of transient and stable expression of foot-and-mouth disease virus capsid proteins in mammalian cells

Foot-and-mouth disease is a highly contagious disease that produces severe economic losses in the livestock industry. This disease is being controlled by the use of an inactivated vaccine. However, the use of recombinant empty capsids as a subunit vaccine has been reported to be a promising candidate because it avoids the use of virus in the vaccine production. A plasmid containing the capsid precursor P12A and protease 3C sequences of foot-and-mouth disease virus (FMDV) was constructed and used to compare transient and stable expression in mammalian cells. When BHK-21 cells were transfected with the recombinant vector, protease 3C cleaved the capsid precursor P12A into the structural proteins VP0, VP1 and VP3. A sucrose gradient demonstrated that the structural proteins assembled into different subviral particles. Attempts to generate a stable cell line only allowed isolating low-level-expressing clones, probably due to the effect of protease 3C on the cells. Moreover, the recombinant protein yield achieved in transient expression assays was much higher than the one achieved in stable expression assays. Results indicate that mammalian cells are a good strategy to produce recombinant FMDV subviral particles. However, the alternative approach of transient gene expression in scalable systems should be used instead of the standard method that involves the generation of a stable cell line.

Foot-and-Mouth Disease Outbreak at Bageshwori Gaushala,Chitwan,Nepal

Cow being the national animal of Nepal has special place in the society.Religious institutions have since prehistoric time reared and cared for cattle herds and the tradition still continues.The Bageshwori Gaushala(cow shelter),initiated by the Hindu Rastraguru Yogi Narhaharinath in mid-1990s is one of the largest of such nucleus indigenous cattle herds in Nepal.Indigenous cattle conservation strategy of Nepal has recognized the importance of this nucleus herd,kept by grazing in the surrounding forest.Ethnoveterinary approaches of managing the herd health is in practice and vaccination was never done.Occasional outbreak did happen in past but overall the herd never suffered severe impact.In April 2018,an outbreak of foot-and-mouth disease(FMD)occurred at Bageshwori,Gaushala,Devghat,Bharatpur Metropolitan-1,Chitwan district of Nepal.In the bovine herd studied,80(53.33%)out of 150 affected animals presented characteristics FMD symptoms as vesicular lesions,drooling salivation,foot lameness,high fever and weakness.Following prompt supportive treatment,the condition improved.The complete recovery of infected herd was observed after 45 d.Laboratory analysis showed the presence of Aphthovirus serotype O in the clinical sample.The present study attempts to provide fresh insights into the recent FMD outbreak,clinical signs,their effective treatment protocol and preventive measures that must be practiced following FMD outbreak.

Chinese herbal medicine for hand-foot-and-mouth disease in children: An overview of systematic reviews

Objective:To summarize the use of Chinese herbal medicines (CHM) for the treatment of hand-foot-and-mouth disease (HFMD) in children and to provide high-level evidence for clinical decision-making.Methods:We conducted an overview of systematic reviews (SRs).Two English-language and four Chinese-language electronic databases were searched from inception to March 31,2018.Published SRs and meta-analyses evaluating CHM use in children with HFMD and reporting clinically-relevant outcomes such as time to fever resolution were eligible for inclusion in this overview.Reviews were accepted if the intervention featured CHM with or without other treatment.Two authors evaluated the methodological quality of the included SRs by using ASMTAR and ROBIS.Results:Thirty SRs comprising 90 244 children with HFMD were included in this overview of SRs.All SRs were published in Chinese between 2011 and 2017.All intervention arms received CHM with or without conventional treatment and were compared against control arms receiving no treatment,conventional treatment alone,or placebo.All 30 reviews reported the time to fever resolution and results demonstrated that CHM had a greater benefit in reducing fever compared with controls.Twenty-three reviews reported the time to rash resolution which presented similar results.Fourteen SRs reported the time to oral ulcer healing and CHM demonstrated a significantly reduced time to resolution compared to controls.The quality of the included SRs was low to moderate as assessed by the AMSTAR tool.Conclusion:Published reviews demonstrated potential benefits of CHM in children with HFMD.Overall,the methodological quality of reviews included in this overview of SRs was low and our findings should be interpreted with caution.We would strongly recommend that future SRs be designed and reported rigorously following PRISMA in order to provide more robust evidence on which to base clinical guidance.

Effects of lonicera granules + symptomatic treatment on serum indexes and immune function in children with hand-foot-and-mouth disease

Objective:To investigate the effects of lonicera granules + symptomatic treatment on serum indexes and immune function in children with hand-foot-and-mouth disease.Methods: A total of 120 children with HFMD who were treated between April 2016 and September 2017 were divided into the control group (n=60) and lonicera granules group (n=60) by random number table method. Control group received clinical symptomatic treatment for hand-foot-mouth disease, lonicera granules group received symptomatic treatment combined with lonicera granules therapy, and the treatment lasted for 7 d. The differences in serum levels of inflammatory factors, myocardial enzyme spectrum indexes and immunoglobulin were compared between the two groups before treatment and after 7 d of treatment.Results: Before treatment, there was no statistically significant difference in serum levels of inflammatory factors, myocardial enzyme spectrum indexes and immunoglobulin between the two groups. After 7 d of treatment, serum inflammatory cytokines IL-6, IL-10, IL-12 and TNF-α levels of lonicera granules group were lower than those of control group;serum myocardial enzyme spectrum indexesα-HBDH, LDH, AST and cTnⅠ levels were lower than those of control group;serum immunoglobulin IgA, IgG and IgM levels were higher than those of control group.Conclusion: Symptomatic treatment combined with lonicera granules therapy can further inhibit the systemic inflammatory response, protect the myocardial cells and enhance the immunity of children with HFMD.

Amplification and Characterization of Bull Semen Infected Naturally with Foot-and-mouth Disease Virus Type Asia1 by RT-PCR

To investigate the security of semen biologically, 15 bull semen samples were collected (of which 5 exhibited clinical signs of Foot-and-mouth disease) and identified by RT-PCR and virus isolation. The results indicated that the semen of the infected bulls were contaminated by Foot-and-mouth disease virus (FMDV), but FMDV was not detected in semen samples from those bulls not showing clinical signs of Foot-and-mouth disease (FMD). This is the first report of the presence of FMDV in bull semen due to natural infection in China. The analysis of the partial sequence of the VP1 gene showed that the virus strain isolated from semen has 97.9% identity with the virus isolated from vesicular liquid of infected bulls showing typical signs of FMD and belonged to the same gene sub-group.

B Cell Epitopes within VP1 of Type O Foot-and-mouth Disease Virus for Detection of Viral Antibodies

In this study,the coding region of type O FMDV capsid protein VP1 and a series of codon optimized DNA sequences coding for VP1 amino acid residues 141-160(epitope1),tandem repeat 200-213(epitope2(+2)) and the combination of two epitopes(epitope1-2)was genetically cloned into the prokaryotic expression vector pPROExHTb and pGEX4T-1,respectively.VP1 and the fused epitopes GST-E1,GST-E2(+2)and GST-E1-2 were successfully solubly expressed in the cytoplasm of Escherichia coli and Western blot analysis demonstrated they retained antigenicity.Indirect VP1-ELISA and epitope ELISAs were subsequently developed to screen a panel of 80 field pig sera using LPB-ELISA as a standard test.For VP1-ELISA and all the epitope ELISAs,there were clear distinctions between the FMDV-positive and the FMDV-negative samples.Cross-reactions with pig sera positive to the viruses of swine vesicular disease virus that produce clinically indistinguishable syndromes in pigs or guinea pig antisera to FMDV strains of type A,C and Asia1 did not occur.The relative sensitivity and specificity for the GST-E1 ELISA,GST-E2(+2),GST-E1-2 ELISA and VP1-ELISA in comparison with LPB-ELISA were 93.3%and 85.0%,95.0%and 90%,100%and 81.8%,96.6%and 80.9%respectively.This study shows the potential use of the aforementioned epitopes as alternatives to the complex antigens used in current detection for antibody to FMDV structural proteins.

Single amino acid substitution of VP1 N17D or VP2 H145Y confers acid-resistant phenotype of type Asia1 foot-and-mouth disease virus

Infection by foot-and-mouth disease virus(FMDV) is triggered by the acidic pH in endosomes after virus uptake by receptor-mediated endocytosis. However, dissociation of the FMDV 146S particle in mildly acidic conditions renders inactivated foot-and-mouth disease(FMD) vaccines much less effective. Type Asia1 FMDV mutants with increased resistance to acid inactivation were selected to study the molecular basis of viral resistance to acid-induced disassembly and improve the acid stability of FMDV. Sequencing of capsid-coding regions revealed four amino acid replacements(VP1 N17D, VP2 H145Y, VP2 G192D, and VP3 K153E) in the viral population of the acid-selected 10th passage. We performed single or combined mutagenesis using a reverse genetic system, and our results provide direct experimental evidence that VP2 H145Y or VP1 N17D substitution confers an acid-resistant phenotype to type Asia1 FMDV.