口蹄疫疫苗研究进展

接种疫苗是特异性预防口蹄疫的有效手段,安全有效的疫苗是成功预防、控制乃至最终消灭口蹄疫的先决条件。在口蹄疫的防控中,灭活疫苗功不可没,但同时也存在许多隐患。随着分子生物学、遗传学及免疫学等学科的迅猛发展,口蹄疫疫苗的研究也取得了较快的进展,各种基因工程疫苗如亚单位疫苗、合成肽疫苗、重组活载体疫苗、核酸疫苗等不断涌现,为口蹄疫的防控带来了新的希望。现对口蹄疫灭活疫苗和新型疫苗的发展过程及其研究进展做一介绍。     

Temporal-spatial epidemiology of foot-and-mouth disease outbreaks in Mongolia, 2000 – 2002

Prior to 2000, foot-and-mouth disease (FMD) had not been observed in Mongolia since 1973; however, between April 2000 and July 2002, Mongolia reported 44 FMD outbreaks that affected cattle, sheep, goats, and camels. The objectives of this study were to describe the distributions of the 44 reported FMD outbreaks in Mongolia and to assess their spatial clustering and directions of movement. Official reports were collected to obtain the number and species of animals both affected and at risk, and the date and geographical coordinates of each outbreak. Significant global and local spatial clusters of reported FMD outbreaks were identified. Disease spread during the second epidemic moved 76° northeast and the spread of the disease during the third epidemic moved 110° northwest. FMD outbreaks were clustered intensely close to other FMD-positive counties. These findings can be used in the future to help plan prevention and control measures in high risk areas.     

猪O型口蹄疫基因工程疫苗候选株的构建及其免疫原性分析

近年来,O型口蹄疫病毒已演化出多种谱系.目前应用的疫苗已不能有效保护多谱系口蹄疫的流行,这给我国猪口蹄疫的防控带来了极大的困难.为了进一步发展免疫原性好、抗原谱广的猪O型口蹄疫疫苗候选株,本研究以O/HN/93现用疫苗毒株的感染性克隆为骨架,构建了含VP3(58位)和VP1(43、48、137、139、140、141和142位)氨基酸改造的全长克隆.线性化的全长cDNA和T7RNA聚合酶质粒共转染BHK细胞后获得拯救病毒.RT-PCR和乳鼠致病性试验表明拯救病毒遗传稳定,具有与亲本病毒相似的致病性.用拯救的基因工程病毒灭活疫苗免疫猪28 d后分别用中国谱系、泛亚谱系和缅甸98谱系猪源毒的流行毒攻击,结果均获得了完全保护(16/16).O/HN/93灭活疫苗免疫猪能完全保护泛亚谱系和缅甸98谱系猪源病毒的攻击(16/16),但不能完全保护(12/16)中国谱系猪源病毒的攻击.结果表明基因工程病毒制备的灭活疫苗提高了对中国型猪源谱系病毒的免疫保护,拓展了抗原谱,是具有良好开发前景的疫苗候选株.     

动物隐性携带口蹄疫病毒的检测方法研究进展

动物隐性带毒的检测在口蹄疫预防控制中有着十分重要的意义。多年来,为了有效地检测出动物隐性携带的口蹄疫病毒,人们采用了各种各样的检测方法,常用的有采集食管-咽喉分泌液进行病毒分离和应用血清学技术检测血清抗体等。近年来,随着分子生物学技术和基因工程技术的发展,不管是检测抗原还是检测抗体,对于口蹄疫隐性带毒的检测和研究都有了一些新的进展,如应用PCR技术检测病毒核酸和应用血清学技术检测抗非结构蛋白抗体,这些新的检测方法的建立使口蹄疫隐性带毒的检测更加可靠。文章对一些口蹄疫隐性带毒的检测方法和最新的研究进展进行了综述。     

RT-PCR evaluation for identification and sequence analysis of foot-and-mouth disease serotype O from 2006 to 2007 in Punjab, Pakistan

FMD clinically positive 250 tissue samples (mouth and hoof epithelium and vesicle swabs, tongue tissue) and 175 secretion samples (milk, saliva, serum, plasma) were evaluated by RT-PCR for the diagnosis of FMD with different pair of universal and serotype-specific primers from 2006 to 2007 in Punjab, Pakistan. Universal primer pair P1/P2 from VP1 gene detected FMD in 182 out of 250 (72.8%) tissues and 92 out of 175 (52.6%) secretion samples, while universal primer 1F/1R from 5′UTR region detected FMD in 218 out of 250 (87.2%) tissues and 142 out of 175 (81.1%) secretion samples. 1F/1R proved better than the P1/P2 primer pair for primary diagnosis of FMD, direct from the clinical positive samples. Direct sequencing of the universal primer pair P1/P2 revealed that O serotype of FMD was circulating in this region. O serotype of FMD was detected with O-1C(ARS4)/PNK 61, AU(O)/AU(rev), AU(O)/PNK61 primer pairs, these primer pairs also compared with each other. AU(O)/AU(rev) and AU(O)/PNK61 detected O serotype of FMD in 88.9% tissue and swab (mouth and hoof vesicle swabs) samples and 71.9% different secretion (milk, saliva, serum, plasma) samples, while O-1C(ARS4)/PNK 61 detected 48.1% tissue and swab (mouth and hoof vesicle swabs) samples and 37.5% different secretion (milk, saliva, serum, plasma) samples. AU(O)/AU(rev), AU(O)/PNK61 primer pairs detected 40.8% more tissue and swab samples, while these pairs detected 34.4% more secretion samples. Cloning of PCR product of AU(O)/AU(rev) VP1 gene and sequencing for phylogenetic studies revealed that O serotype of FMD circulating in Punjab, Pakistan was genetically very diverse from the ‘O’ serotype in Middle East and Europe. The dendrogram showed that Pakistan ‘O’ serotype was very much similar genetically to its neighbor countries (Sri Lanka, India, Iran, Iraq, and China) and PanAsia 1 lineage which caused 2001-outbreak in UK and 1994-outbreak in Saudi Arabia, etc.     

Spatial distribution of foot-and-mouth disease in Pakistan estimated using imperfect data

We estimated the spatial distribution of foot-and-mouth disease (FMD) in Pakistan; we used a probability co-kriging model and the number of FMD outbreaks reported between 1996 and 2000 by Pakistan to the Office International des Epizooties. We used a k-Bessel model and small-ruminant and human densities as surrogate covariates for the population at risk and for livestock markets and movements, respectively. Compared to no or only one covariate, the co-kriging model with both densities provided the best fit to independently obtained data on the spatial distribution of virus isolations (P = 0.57). The estimated probability of an FMD outbreak per 25 km² cell ranged from 0.017 to 0.812, with the maximum relative probability of 47.8 (0.812/0.017). Areas with the highest relative probability of having an FMD outbreak were located in the Punjab region; this is a major animal-production area located along a traditional international animal-trade route.     

口蹄疫综合防控与环境控制问题

口蹄疫是由口蹄疫病毒引起的一类急性、热性、高度接触性传染病,对世界各国养殖业造成了严重的经济损失.目前通过环境控制的策略来综合防控口蹄疫引起了人们的重视.论文就口蹄疫的流行概况、综合防控及环境控制与口蹄疫的防控相互关系进行探讨,旨在为口蹄疫的综合防控提供参考.     

口蹄疫转基因植物疫苗研究进展

口蹄疫是国际社会广泛关注的严重动物传染病之一,其控制措施主要是疫苗免疫。传统疫苗在该病的防控中起了重要的作用,但是其弊端也不容忽视,研制新型的口蹄疫疫苗是今后的发展方向。近几年来,随着生物技术的发展,可饲疫苗由于其具备的诸多优点而成为国内外研究的热点,口蹄疫转基因植物疫苗的研究在国内外也已经开始,并取得了一定的成果,期望易感动物食用此转基因植物后产生免疫力,达到预防口蹄疫的目的。文章综述了国内外口蹄疫转基因植物疫苗研究的成果。     

Foot-and-mouth disease. Challenge of cattle after multiple vaccinations

Four groups of six cattle were vaccinated from two to five times at 6 month intervals with two different trivalent FMD vaccines licensed in the given year. The FMDV type A strains in the vaccines designated A5F and A5B were closely related. Three months after the last vaccination the cattle were challenged by contact with animals inoculated with the original field strain A5B. The inoculated animals developed typical FMD symptoms with vesicles in the mouth and on the feet. Those cattle which had received vaccines that did not contain strain A5B also became severely sick, even after five vaccinations. Animals vaccinated twice with type B containing vaccine were also not completely protected. A safe protection can obviously only be achieved for fairly short periods of time if vaccine and challenge strain are homologous. It is proposed to change the rules of licensing, to speed up the procedure to vaccinate in cases of outbreaks. The need for further research, especially into improving vaccines, is stressed.     

Risk factors for foot-and-mouth disease in Zambia, 1981–2012

The aim of this study was to describe the spatial distribution of foot-and-mouth disease (FMD) outbreaks in Zambia for the period January 1981–December 2012 and to quantify the association between geographical features (proximity to roads, national parks, wetland areas) and the spatial distribution of FMD using a Poisson point process model.     

国内现有猪口蹄疫O型灭活疫苗对2010年流行毒株的抵抗力

在2010年上半年,猪O型口蹄疫不断发生,作为国内几个生产厂家的猪口蹄疫现用疫苗种毒的提供者,我组进行了猪源流行毒对该种毒灭活疫苗免疫猪的攻毒保护试验及观察。结果如下:1)、猪源流行毒致猪发病速度慢。猪源流行毒接种猪后,致猪发病的时间在3～5天,而疫苗株致猪发病时间是接种后2～3天;2)、该疫苗毒株能有效抵抗猪源流行毒的攻击,PD50大于6;3)、免疫保护和ELISA抗体水平没有线性相关性;4)、免疫保护与140S抗原量有相关性。     

Microarray-based detection and typing of foot-and-mouth disease virus

Foot-and-mouth disease virus (FMDV) is the most economically important veterinary pathogen because of its highly infectious nature and the devastating effects the virus has on the livestock industry. Rapid diagnostic methods are needed for detection and typing of FMDV serotypes and differentiation from other viruses causing vesicular diseases. We developed a microarray-based test that uses a FMD DNA chip containing 155 oligonucleotide probes, 35-45 base pair (bp) long, virus-common and serotype-specific, designed from the VP3-VP1-2A region of the genome. A set of two forward primers and one reverse primer were also designed to allow amplification of approximately 1100 bp of target sequences from this region. The amplified target was labelled with Alexa-Fluor 546 dye and applied to the FMD DNA chip. A total of 23 different FMDV strains representing all seven serotypes were detected and typed by the FMD DNA chip. Microarray technology offers a unique capability to identify multiple pathogens in a single chip.     

Foot-and-mouth disease: a review of intranasal infection of cattle, sheep and pigs

In an outbreak of foot-and-mouth disease (FMD) it is important to identify animals at risk from airborne virus. Investigations have been carried out over the years to determine the dose required to infect cattle, sheep and pigs by the intranasal route. This paper reviews the results of investigations for animals which have been infected by instillation or spraying a virus suspension into the nostrils or by exposure to affected animals through a mask or by indirect contact. The lowest doses were found by use of a mask. With virus from affected pigs given through a mask, doses of 18 infectious units (IU) in cattle and 8 IU in sheep were found to cause infection and give rise to lesions. Overall, cattle required the least amount of virus followed by sheep. Pigs required a dose of 22 IU to cause infection and a dose of 125 IU to give rise to lesions. In many experiments pigs failed to become infected. With all three species the dose varied with the individual animal and the virus strain. For modelling previous outbreaks and in real time, a dose of 8 IU or 10 and 50% infectious doses (ID50) could be used where cattle and sheep were involved. Experience in the field, combined with the results from experiments involving natural infection, indicate that pigs are not readily infected by the intranasal route. However, for modelling purposes a dose of about 25 IU should be used with care. Investigations are needed to determine doses for virus strains currently in circulation around the world. In addition, the nature of the aerosol droplets needs to be analysed to determine how the respective amounts of infective and non-infective virus particles, host components and, in later emissions, the presence of antibody affect the survival in air and ability to infect the respiratory tract. Further work is also required to correlate laboratory and field findings through incorporation of the doses into modelling the virus concentration downwind in order that those responsible for controlling FMD are provided with the best available assessment of airborne spread. Finally, the doses found for infection by the intranasal route could be applied to other methods of spread where virus is inhaled to assess risk.     

A survey to investigate movements off sheep and cattle farms in New Zealand,with reference to the potential transmission of foot-and-mouth disease

AIM: To quantify the numbers and extent of movements off sheep and cattle farms in New Zealand, in order to construct more realistic simulation models to investigate how infectious diseases such as foot-and-mouth disease (FMD) might spread.

METHODS: Farmers from 500 randomly selected farms, comprising 100 from each of the following sectors, viz beef, dairy, grazing/dairy heifer rearing, sheep, and mixed sheep and beef, were asked to fill in diaries in which they recorded the movements of all animals, products, people, vehicles and equipment coming on to or leaving their farms during two separate 3-week periods, representing relatively ‘busy’ and ‘quiet’ times of the year with respect to livestock movements. Where possible, the destination of each movement was identified and geo-coded, to allow the distance travelled to be calculated. Each movement was then classified according to the risk of transfer of FMD virus (FMDV), should the disease have been present on the study farm at the time of the movement. The data were then analysed to establish movement frequencies and distributions of distances travelled, by the different pastoral livestock sectors.

RESULTS: Two hundred and seventeen farmers returned one or more diaries. One hundred and ninety-three farmers completed a Busy-period diary, recording a total of 12,052 movements off their farms, a crude average of 62.4 per 3-week period, or 2.97 per day. Of these, 4.0% involved the transport of livestock, equating to 0.12 livestock consignments per day. In contrast, 186 Quiet-period diaries were returned, recording a total of 10,885 movements off, representing a crude average of 58.5 during the 3-week period, or 2.78 per day. Of these, 2.1% involved livestock, equating to 0.06 livestock consignments per day. The mean and median distances travelled during the Busy periods were 30.9 km and 13.1 km, respectively (range 0–1,167 km). In comparison, the mean and median distances travelled during Quiet periods were 41.3 and 14 km, respectively (range 0.4–1,203 km).

CONCLUSIONS: People, vehicles, livestock and other items can travel off pastoral livestock farms in New Zealand to other farms either directly or via saleyards over extensive distances. This has implications for the potential spread of infectious diseases such as FMD. Movement parameters intended for use in the InterSpread Plus inter-farm simulation model of FMD were established, which will facilitate the prediction of likely spread and efficacy of controls in the unlikely event of a real-life outbreak.     

Use of the scan statistic on disaggregated province-based data: foot-and-mouth disease in Iran

The spatial scan statistic was applied to density-smoothed data that approximated the spatial distribution within the area and reduced the potential bias produced when location data have been aggregated for large areas. The method is illustrated, using data on the location of foot-and-mouth disease (FMD) outbreaks in Iran. Data examined were 4477 FMD outbreaks reported on a per province basis between June 1996 and September 2003. A kernel density of the outbreak locations was estimated, using a fixed radius and the centroid of each province as the designated location of all cases reported for the province. The radius that produced a density map with the highest correlation with expert opinion was 4° (latitude/longitude). Livestock density was used as a proxy for the underlying population at risk of acquiring FMD. Livestock and outbreak density maps were overlain to obtain the number of outbreaks and livestock in each of 15,599 cells covering the mapped surface of the country. A spatial scan statistic was applied to the density-smoothed data assuming that the outbreaks had a Poisson distribution. Results were compared with those obtained using a spatial scan statistic on provincially aggregated data. Application of the spatial scan statistic on the density-smoothed data allowed identification of clusters (P < 0.01) related more to the actual geographic distribution of cases (expert opinion) and of animals at risk, than to the distribution of the provinces. Significant clusters of FMD were identified that coincided with roads, neighboring countries, and high-density population areas, suggesting that the region may represent a route for cross-continent transmission of FMD.     

猪口蹄疫的综合防控

口蹄疫是由口蹄疫病毒(Foot-and-mouth disease virus, FMDV)感染引起偶蹄动物共患的急性、热性、高度接触性传染病,最易感染的动物是黄牛、水牛、猪、骆驼、鹿、羊等;黄羊、麝、野猪、野牛等野生动物也易感染此病。世界动物卫生组织将其列为A类动物传染病,我国将其列为I类动物疫病。文章从口蹄疫病原的特性、流行病学、临床特征及病理变化等方面对口蹄疫的防控措施进行了总结、阐述。     

口蹄疫灭活疫苗研究进展

灭活疫苗仍然是当前口蹄疫免疫控制的主要疫苗,研制口蹄疫灭活疫苗具有重要的现实意义,其研制方法在不断丰富和完善。文章详细描述了口蹄疫灭活疫苗研制的重要步骤及其进展,包括毒株筛选,病毒生产、灭活、抗原浓缩、纯化和配苗及疫苗的质量控制。制苗毒株的选择是研制疫苗的首要问题,直接关系到疫苗的免疫效果;病毒灭活和随后的安全试验是在制备灭活FMD疫苗中最关键的步骤;抗原浓缩纯化和配苗是保证疫苗良好免疫效果的必需。最后阐述了口蹄疫灭活疫苗在口蹄疫防控中的作用。     

Village level risk factors for foot-and-mouth disease in Northern Thailand

A study was undertaken in northern Thailand to identify factors which put some villages at higher risk of foot-and-mouth disease (FMD) outbreaks than others. The number of FMD outbreaks experienced in the previous 5 years and data on 145 putative risk factors were obtained by interview from 60 villages during 1991–1992. Univariable analyses identified 27 factors for further investigation using logistic regression. When villages were classified into three FMD frequency groups of zero to one, two to three or four or more outbreaks in the last 5 years, the important factors explaining the differences in risk were the total number of cattle and buffaloes purchased in the previous year, the number of neighbouring villages which shared a common water source and whether agriculture was the most important source of cash income for the village. These factors were also the most important variables in explaining the difference in risk when comparing villages with zero or one outbreak with those having four or more. We concluded that the greatest impact on reducing spread of FMD among villages would be obtained through the development of strategies to reduce the likelihood of introduction through livestock purchases and for villagers to take greater care when livestock are grazed with those from neighbouring villages and when sharing common water supplies.     

口蹄疫病毒免疫学研究进展

口蹄疫病毒(FMDV)是小RNA病毒科、口蹄疫病毒属成员,其感染后引起的口蹄疫是一种严重危害畜牧业发展的传染病,并对经济发展、国家声誉及国际关系造成重要影响.论文就FMDV的主要抗原位点、FMDV的细胞受体、机体对FMDV抗原的免疫反应、FMDV抗原的变异与进化等做一综述.     

A dynamic, optimal disease control model for foot-and-mouth disease: I. Model description

A dynamic optimization model was developed and used to evaluate alternative foot-and-mouth disease (FMD) control strategies. The model chose daily control strategies of depopulation and vaccination that minimized total regional cost for the entire epidemic duration, given disease dynamics and resource constraints. The disease dynamics and the impacts of control strategies on these dynamics were characterized in a set of difference equations; effects of movement restrictions on the disease dynamics were also considered. The model was applied to a three-county region in the Central Valley of California; the epidemic relationships were parameterized and validated using the information obtained from an FMD simulation model developed for the same region. The optimization model enables more efficient searches for desirable control strategies by considering all strategies simultaneously, providing the simulation model with optimization results to direct it in generating detailed predictions of potential FMD outbreaks.