套式PCR在猪流行性腹泻病毒感染病例诊断中的应用

猪流行性腹泻是对养猪业危害较大的一种疾病,导致腹泻的原因多种多样,给临床诊断带来了困难。本研究将套式PCR技术成功应用于猪流行性腹泻临床病例的快速诊断中,旨在为猪流行性腹泻临床病例的快速鉴别诊断提供一种新的思路。     

猪流行性腹泻病毒ORF3和M基因的克隆与序列分析

为了解福建省猪流行性腹泻病毒(Porcine epidemic diarrhea virus,PEDV)毒株ORF3和M基因变异情况,本研究从4个不同规模猪场发生仔猪“顽固性腹泻”疾病的病料中扩增到4株PEDV的ORF3和M基因,并进行了序列分析.结果表明:4株PEDV ORF3基因均含有675个碱基,编码224个氨基酸,与GenBank中登录的代表性PEDV核苷酸的同源性为89.7％～100.0％,氨基酸的同源性为94.7％～99.6％,其中FJPT毒株核苷酸的同源性与CV777 truncated毒株最低,仅为89.7％,FJNP毒株核苷酸的同源性与中国北方流行的CH ZKFG 11毒株最高,达100.0％.M基因含有681个碱基,编码226个氨基酸,与GenBank中登录的代表性PEDV核苷酸的同源性为94.8％～100.0％,氨基酸的同源性为94.8％～99.6％,其中FJPT毒株核苷酸的同源性与日本D89752毒株最低,仅为94.8％,FJNP毒株核苷酸的同源性与韩国CPF193、泰国毒株最高,达100.0％.4株PEDV毒株均与国内外流行强毒株的亲缘关系较近,与attenuate DR13弱毒株的亲缘关系较远.     

猪两种腹泻病毒混合感染的病原诊断

对我省流行的疑似猪病毒性腹泻进行了病原诊断，通过细菌培养、ＲＮＡ电泳和电镜形态观察等辅助诊断和免疫萤光特异性诊断，确诊该病病原为猪传染性胃肠炎病毒（ＴＧＥＶ）与猪流行性腹泻病毒（ＰＥＤＶ）的混合感染，混合毒代号为ＴＰＶ３。     

Classical swine fever in The Netherlands 1997-1998: a description of organisation and measures to eradicate the disease

The central and regional organisation of the campaign to eradicate the CSF epidemic in the Netherlands in 1997/1998 is described. The main instruments used in the campaign were based on stamping-out and movement restrictions specified by the European Union. Additional instruments were used for the first time, namely, pre-emptive culling of contact and neighbouring farms, compartmentalisation of transport, monthly serological screening in established surveillance areas and supervised repopulation of all farms in the former surveillance zone. Two other measures, the killing of very young piglets and a breeding ban were introduced to reduce production in established surveillance zones. Several factors complicated the eradication campaign, for instance, the late detection of the first infection; artificial insemination as a source of infection; the organisation of pig farming in the Netherlands, with its highly concentrated production and dependence on the transport of stock from one unit to another; insufficient rendering capacity; decreasing sensitivity of clinical inspection; and extremely high costs.     

Identification and Molecular Characteristics Analysis of Porcine Epidemic Diarrhea Virus Variants

To ascertain a diarrhea case in a pig farm in Shandong province,the pathological changes of dead piglets were observed and nested RT-PCR test was carried out on 7 diarrhea samples for porcine epidemic diarrhea virus (PEDV).Pathological examination revealed that intestine was detected as enlargement,hyperemia and edema.After histological examination,the typical microscopic lesions of intestine were disappearance of epithelial cells,villus shrinkage and shortening.The result of nested RT-PCR showed that all of the 7 samples could amplify a specific target band of PEDV.Molecular characteristics of two field strains showed that they had an amino acid homology of 92.3% to 92.4% with vaccine CV777 and 96.6% to 98.6% with other previous field strains which sequences were downloaded from GenBank.Phylogenetic tree analysis further revealed that all of PEDV strains could be mainly divided into two clusters of G1 and G2. G2 consisted of the field strains of our study and other field strains from USA,China and so on,which had the same sequence characteristics of two insertions and one deletion,while G1 consisted of all vaccines of CV777 and several older field strains from China and Korea.These results indicated that our field strains were the dominant strains in recent epidemic diarrhea occurrence.Moreover,its molecular characteristics might be a characterization of differentiating the field and vaccine strains.     

Tick-borne encephalitis epidemic in Hungary 1951–2021: The story and lessons learned

The authors analysed epidemiological data of the Hungarian tick-borne encephalitis epidemic from the past seven decades. A total of 911 meningitis serosa cases were described from 1930-1950 s by local hospital physicians, indicating that the virus had been present in the country decades before its official identification in 1952. The virus spread freely in the 1950s–1960s, occupying almost all habitats where ticks occurred in large numbers. The increasing number of cases drove authorities to classify this illness as a notifiable disease in 1977 and to organize the first measures to stop the epidemic. Statistical analysis revealed that the large-scale vaccination launched from the 1990s was responsible for the sharp decrease in the number of human cases from 1997. A significant negative correlation was found between the number of vaccine doses sold and human cases 6 years later. The TBEV endemic area covers 16.57% of the territory and 16.65% of the population of the country. In the last 10 years, 186,000 vaccine doses/year in average were enough to keep the incidence of human TBEV infections between 0.45 and 0.06/100,000 persons. A 20-year-long study found evidence for easing clinical signs in TBEV-infected hospitalized patients. Statistics found a sharp decrease in the number of samples sent for TBEV diagnosis after 1989. Male dominance of patients was characteristic of the epidemics since the 1940s, but now analysis of detailed data from the 1981–2021 period (60.5%–87.5%) proved the statistical significance of this dominance. Obviously, the voluntary vaccination programme was the tool which broke the spread of the epidemic. Widespread public awareness of the disease and the tick vector, probable evolutionary spread of less pathogenic virus strains supplemented with the vaccination campaign led to a negligible level of human TBE cases in Hungary in the last years.     

Etiological Investigation of Diarrhea in Newborn Piglets and Molecular Characterization and Genetic Variation Analysis of ORF3 Gene of PEDV in North Guangdong

In order to understand the main pathogen of newborn piglets diarrhea in North Guangdong region, 31 diarrhea samples were collected from six pig farms in North Guangdong, the pathogen of porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV) and porcine rotavirus (PoRV) were detected by Real-time RT-PCR, meanwhile, ORF3 gene of PEDV amplified from positive samples were sequenced and analyzed. Pathogen detection results showed that 83.87%(26/31) samples were positive for PEDV, all herds and samples were negative for TGEV and PoRV. The sequence analysis revealed that the ORF3 gene of 5 epidemic strains of PEDV were all 675 bp, homologies of nucleotides were 98.7% to 100.0%, and homologies with reference sequences of nucleotides were 94.5% to 100.0%, and some gene mutation in common nucleotides site. The results of gene phylogenetic trees showed that PEDV could be divided into two groups, PEDV genetic relationship between field strains in North Guangdong and some regions in China, Southeast Asia, North America, Europe from 2013 to 2015 was closer, classed as a gene subgroup, from 2011 to 2012 main epidemic strains in our country and vaccine strains classed as other two gene subgroups. These results indicated that PEDV infection was the main pathogen of newborn piglets diarrhea in North Guangdong region, as time passed, the PEDV epidemic strains gene presented a tendency of evolution and variation.     

河南省部分地区猪流行性腹泻病毒ORF3和N基因的克隆及其遗传进化分析

为探究河南省猪流行性腹泻病毒部分毒株的遗传进化情况,采用RT-PCR对2017年2月至2018年1月在河南省部分地区猪场收集到的25份PEDV阳性病料进行ORF3和N基因的扩增,并对其进行克隆、序列比对及遗传进化分析。结果显示,PEDV毒株的ORF3基因序列是由675个核苷酸组成的,与经典毒株CV777之间核苷酸及氨基酸同源性分别为95.2%~97.5%和95.1%~96.9%。N基因之间的核苷酸与氨基酸同源性分别为96.2%~100%和93.8%~99.8%;与经典毒株CV777核苷酸与氨基酸的同源性分别为94.7%~95.8%和93.2%~96.8%。河南部分地区PEDV流行毒株与经典毒株CV777不在同一分支,说明猪场暴发猪流行性腹泻与免疫接种疫苗后依旧难以控制的原因,可能与大多数PEDV河南流行株发生变异有关。     

非洲猪瘟形势下猪流行性腹泻的防控措施

自2018年第一例非洲猪瘟疫情报道至今已经两年多了.为了快速扩繁猪群,许多清空场及新建场不得不大量外购来源复杂后备猪进群.因此,猪场在对猪流行性腹泻的防控上面临着更大的挑战.文章介绍了病毒性腹泻暴发的原因、防控难点和综合防控措施,期望为养殖业同人在实际生产中,科学防控猪流行性腹泻提供一定的参考借鉴.     

Study on Anti-viral Effect of Rhubarb on Porcine Epidemic Diarrhea Virus in vitro

The inhibitory effects of rhubarb on cell lesion, which was caused by porcine epidemic diarrhea virus (PEDV), and its mechanism had been studied in this research.The maximum safe concentration of rhubarb solution applied to Vero cells was determined through morphological observation, and then the PEDV infected Vero cell model in vitro was established, by which the inhibitory effect of rhubarb on virus was studied.The experimental method had also been established based on this cell model.The result showed that the maximum safe concentration was 3.28 mg/mL.Both morphological observation and MTT assay were indicated that rhubarb could not only block the attachment of the virus to Vero cell and inhibit the synthesis and reproduction of the virus, but also could directly inactivate the virus.This research showed that rhubarb had an inhibitory effect on PEDV, and provided a theoretical basis for the prevention and treatment of clinical porcine epidemic diarrhea.