Estimation Models for the Morbidity of the Horses Infected with Equine Influenza Virus

Estimation formulas for the morbidity of horses infected with equine influenza virus by linear regression, logistic regression and probit transformation were developed, using data from the outbreak at the Sha Tin Racing Track in Hong Kong in 1992. Using these formulas, we estimated the equine influenza virus morbidity rates at training centers belonging to the Japan Racing Association (JRA) in October 1997 and in October 1998. In 1998 JRA started a new vaccination program, and every horse must now be vaccinated twice per year. At that time, the vaccine included two US lineage virus strains, the A/equine/Kentucky/81 strain and the A/equine/La Plata/93 (LP93) strain, against equine type-2 influenza viruses; it did not include any EU lineage virus strains, such as A/equine/Suffolk/89 (SF89). Comparing the geometric mean (GM) values of hemagglutination inhibition (HI) titers between the LP93 strain and the SF89 strain in 1997 and in 1998, they both rose significantly at every age (p<0.05) by Wilcoxon test. Calculations by the simulation models show the morbidity rates for LP93 diminished from 0.439 (linear), 0.423 (logistic) and 0.431 (probit) to 0.276 (linear), 0.265 (logistic) and 0.271 (probit), respectively. On the other hand, the estimated morbidity rates for SF89 diminished only slightly from 0.954 (linear), 0.932 (logistic) and 0.944 (probit) to 0.946 (linear), 0.914 (logistic) and 0.927 (probit), respectively. Our simulation models could estimate the effect of the vaccine on each of the equine virus strains represented by the morbidity of infected horses. Thus, they are useful for vaccine evaluation.     

野马鼻肺炎病毒的分离鉴定

从新疆昌吉州吉木萨尔野马饲料养繁殖中心送检的野马病料中分离到１株病毒,我们对其进行了鉴定。该病毒株在ＢＨＫ－２１细胞上连传６代出现典型的细胞病变,用适应ＢＨＫ－２１细胞的病毒接种鸡胚成纤维细胞,乳豚鼠肾原代细胞,豚鼠睾丸细胞均出现程度不同的细胞病变。     

Comparison of the Virucidal Effects of Disinfectant Agents Against Equine Influenza A Virus

Disinfection is one of the most important biosecurity measures to minimize disease spread during outbreaks of equine influenza. Although many disinfectants are commercially available, information about their effects against equine influenza A virus (EIV) is limited. This report describes an evaluation of the effects of six disinfectants against EIV (∼10^4.7 egg infectious dose 50/200 μL) under different conditions (reaction time [10 and/or 30 minutes], temperature [4°C–25°C], and the absence and/or presence of uninfected allantoic fluid that served as our best choice of “organic matter” equivalent). Although the efficacy of didecyldimethylammonium chloride decreased with decreasing reaction temperature, the compound showed the highest efficacy of the three quaternary ammonium compounds tested in this study. The effects of sodium dichloroisocyanurate and nonoxynol iodine were not affected by reaction time or temperature, but they were affected by the presence of organic matter. Antec Virkon S containing potassium peroxymonosulfate and sodium chloride consistently inactivated EIV regardless of reaction time, temperature, and the presence of organic matter. These findings will help us to take rational biosecurity measures during outbreaks of equine influenza.     

Phylogenetic and Molecular Characterization of the Equine Influenza Virus A (H3N8) Causing the 1997 and 2004 Outbreaks in Morocco

Reported here are the results of antigenic and genetic characterization of equine influenza strains causing local outbreaks reported in Morocco, respectively, in 1997 and 2004. The antigenic and genetic characterizations of the equine influenza virus H3N8 are reported here. The highest similarity between the HA1 nucleotide sequences of A/equine/Nador/1/1997 and those of A/equine/Rome/5/1991 and A/equine/Italy/1199/1992 demonstrate that A/equine/Nador/1/1997 belongs to the European lineage. On the other hand, A/equine/Essaouira/2/2004 and A/equine/Essaouira/3/2004 were classified in the predivergent lineage. The present work emphasizes the importance of a national influenza survey program, which requires a collaborative laboratory network to promote the collection and characterization (antigenic and genetic) of equine influenza viruses in real time.     

规模化驴场中马流感病毒H3N8亚型感染情况调查

[目的]调查山东省聊城市规模化驴场中马流感病毒的感染情况,并分析其可能的来源。[方法]从聊城的规模化驴场采集病料和血清,通过HI试验检测驴血清中的马流感病毒H3N8亚型抗体的阳性率。使用RT-PCR技术扩增肺脏和鼻腔棉拭子样品中的马流感病毒M基因,对获得的马流感病毒M基因与不同流感病毒的M基因进行序列比对,推测其来源。[结果]HI试验表明,120个血清样品中马流感病毒H3N8亚型血清抗体阳性率为33.3%(40/120);其中,母驴的马流感病毒H3N8亚型血清抗体阳性率为42.5%(17/40)、公驴为32.5%(13/40)、驴驹为25.0%(10/40)。通过RT-PCR检测发现,32.3%(21/65)的样品可测出目的条带。通过序列比对得出,该试验获得的流感病毒M基因与马属动物的H3N8亚型流感病毒高度同源(CY032222、CY032318、CY028821等),同源性最高可达99.8%。[结论]马流感病毒在聊城周边的数个规模化养驴场发生流行。该研究从驴体内分离的流感病毒M基因属于马流感病毒H3N8亚型M基因。     

H3N2亚型禽流感病毒及猪流感病毒感染人肺腺癌细胞后增殖的差异性变化

人肺腺癌细胞A549被H3N2亚型禽流感病毒和猪流感病毒感染后,探讨不同毒株跨种属感染人呼吸道组织的可能性和趋势性。通过观察病毒感染A549细胞后的细胞病变(CPE)、血凝试验(HA)和50%组织细胞感染量(TCID50)来比较不同毒株感染细胞后的复制差异和毒力改变,发现同一亚型不同来源的流感病毒对A549细胞感染和复制能力有较大差异,哺乳动物来源的病毒更有感染人呼吸道细胞的趋势,SW/GX/NS2783/2010有潜在的感染人细胞的能力。感染过程中的细胞因子变化和病毒基因组组成特点是进一步研究的方向,应重点关注具有跨种属感染趋势的流感病毒及其特殊分子决定簇的组成和特点。     

实验性鸡大肠杆菌病病理学动态变化

用致病性大肠杆菌O18分离株和／或低致病性禽流感病毒（Mildly pathogenic avian influenza virus ,MPAIV)接种10－12日龄SPF鸡。在接种后1－96h进行临床症状与大体病理变化、组织学观察发现：大肠杆菌接种组、MPAIV接种组和健康接种组除扑杀鸡外未见鸡死亡，MPAIV与大肠杆菌混合接种组除扑杀鸡外死亡率为24％。混合接种组的病变比大肠杆菌接种组出现的时间早，恢复也慢，各脏器的病理变化更严重。MPAIV主要引起各实质器官的坏死，结果表明，大肠杆菌经气管内接种后试验鸡主要表现为呼吸道的炎症反应；MPAVI可使鸡大肠杆菌病严重化。     

H3N8亚型马流感病毒单克隆抗体的制备及鉴定

以灭活马流感病毒(EIV)A/Equine/Jilin/1/1989(H3N8)为免疫原,免疫Balb/c小鼠,经常规细胞融合后,用血凝抑制试验(H1)和间接ELISA方法筛选获得3株(3C2、5G10和5A10)能稳定分泌H3N8亚型马流感病毒单克隆抗体(mAb)的杂交瘤细胞株.其中3C2和5G10为IgG2α,5A...     

A型流感病毒蛋白研究进展

流感是由A型流感病毒引起的一种感染和／或疾病综合征。A型流感病毒已成为鸟类、人类和低等哺乳动物疾病的主要病原之一，其病毒基因组由8个节段组成，共编码10种以上的多肽，每种多肽都有着重要的生物学功能。     

MPAIV、NDV Lasota株分别与较低致病性禽源E.coli的联合感染试验

以 2× 1 0 5EID50 的低致病性禽流感病毒 ( mildly pathogenic avian influenza virus,MPAIV)、2× 1 0 6 EID50 新城疫病毒 L asota株( Newcastle disease virus Lasota strain,NDVL asota)气管内注射 1 0日龄 SPF鸡 ,2 4h后 ,同剂量、同法重复感染一次 ;48h后 ,分别气管内注射较低致病性禽病原性大肠杆菌 1 2 0( O1 8)和 1 73( O2 6 )株 ,2× 1 0 7CFU/羽 ,2 4h后同剂量、同法重复攻毒一次 ,连续观察 1 0d。结果 :MPAIV单独感染组死亡率为 53% ;NDV Lasota株单独攻毒组未见死亡 ;大肠杆菌 1 2 0株单独攻毒组死亡率为 40 % ,1 73株单独攻毒组死亡率为 7% ;MPAIV与大肠杆菌1 2 0株联合攻毒组的死亡率为 87% ,NDV L a-sota株与 1 2 0株联合攻毒组的死亡率为 40 % ;MPAIV与大肠杆菌 1 73株联合攻毒组的死亡率为 80 % ,NDV Lasota株与 1 73株联合攻毒组的死亡率为 2 0 %。     

猪传染性胃肠炎病毒呼吸道途径侵染后在体内的动态分布

应用猪传染性胃肠炎病毒（TGEV）分别对25日龄和40日龄的SPF仔猪进行气管内注射攻毒。攻毒后每24h致死1头仔猪．用免疫荧光染色检测各脏器内病毒的消长情况。并采血检测抗体。结果．在25日龄的仔猪中．仅3头出现腹泻症状。且很快耐过；剖检可见不同程度的肺部病理变化；攻毒后24h气管和肺脏样品的IFA呈阳性。抗体效价随着时间的延长而增高。40日龄仔猪临床表现和训检变化基本正常。所有的胃肠道样品的IFA均为阴性。结论：气管内注射后，TGEV可在呼吸系统中增殖．但未能在消化道系统中检出。     

自噬与A型流感病毒感染

自噬是广泛存在于真核细胞内的新陈代谢过程,对于维持细胞自稳具有关键作用。在病毒感染过程中,自噬表现出两面性,既可被利用来促进病毒复制,又在宿主抗感染免疫应答中发挥重要作用。论文以严重危害畜禽生产和人类健康的A型流感病毒为例,论述了细胞自噬与病毒感染及在宿主免疫应答过程中的相互关系,旨在进一步深入理解A型流感病毒的致病分子机制,为抗流感病毒感染的相关研究提供参考。     

Antibody Responses Against Equine Influenza Virus Induced by Concurrent and by Consecutive Use of an Inactivated Equine Influenza Virus Vaccine and a Modified Live Equine Herpesvirus Type 1 Vaccine in Thoroughbred Racehorses

An inactivated equine influenza virus (EIV) vaccine and a live equine herpesvirus type 1 (EHV-1) vaccine are usually administered concurrently to Thoroughbred racehorses in Japan. The objective of this study was to evaluate whether concurrent administration of an inactivated EIV vaccine and a live EHV-1 vaccine in Thoroughbred racehorses influences the antibody response against EIV. We compared the antibody response against EIV in horses administered both vaccines on the same day (Group A; n = 27) and the response in horses administered an inactivated EIV vaccine first and then a live EHV-1 vaccine 1–2 weeks later (Group B; n = 20). In both groups, geometric mean hemagglutination inhibition (HI) titers against A/equine/Ibaraki/1/2007 and A/equine/Yokohama/aq13/2010 increased significantly after EIV vaccination. However, the percentage of horses that showed a twofold increase or greater in HI titers against A/equine/Yokohama/aq13/2010 was significantly higher in Group B (75%) than in Group A (37%; P = .02). These results suggest that the concurrent use of an inactivated EIV vaccine and a live EHV-1 vaccine reduced the immune response against EIV to some extent, and it would be better to use these vaccines consecutively, especially for naïve horses or horses whose vaccination history is incomplete.     

马流感病毒双抗体夹心ELISA检测方法的建立

为建立一种快速、有效的检测马流感病毒(Equine influenza virus,EIV)的方法,以EIV中国分离株A/马/新疆/07(H3N8)制备的多克隆抗体为捕获抗体,原核表达的核蛋白(NP)制备的单克隆抗体为检测抗体,在国内首次建立了检测EIV的双抗体夹心ELISA方法.用该检测方法分别检测EIV、马动脉炎病毒、马疱疹病毒1型、马疱疹病毒4型和马乙型脑炎病毒阳性样品.结果表明,该ELISA方法具有良好的特异性;与常规检测EIV的血凝试验相比,其敏感性是后者的2.5～10倍;同时与H7N7亚型EIV有交叉反应.攻毒试验结果表明该方法可有效检测鼻腔分泌物中的EIV.该方法的建立为EIV的检测及早期防控提供了有效工具.     

Prevalence of Influenza A Virus in Exhibition Swine during Arrival at Agricultural Fairs

The exhibition swine at agricultural fairs provides a critical human–swine interface that allows for the bidirectional transmission of influenza A virus (IAV). Previous IAV surveillance at the end of fairs has resulted in frequent detection of IAV‐infected swine; little is known, however, about the frequency with which swine arrive at fairs already infected with IAV. We investigated the IAV prevalence among exhibition swine entering fairs to better understand the epidemiology of IAV in this unique human–swine interface. In 2014, snout wipes were collected from 3547 swine during the first day of nine agricultural exhibitions in Indiana and Ohio. Samples were screened for IAV using rRT‐PCR and positive samples were inoculated into cultured cells for virus isolation. The overall IAV prevalence detected among swine arriving at exhibitions was 5.3% (188/3547) via rRT‐PCR and 1.5% (53/3547) via virus isolation, with IAV being detected and recovered from swine at 5 of the 9 exhibitions. Within the fairs with IAV‐positive swine, the individual exhibition IAV prevalence ranged from 0.2% (1/523) to 34.4% (144/419) using rRT‐PCR and 0.2% (1/523) to 10.3% (43/419) with virus isolation. Single IAV subtypes were detected at three of the fairs but subtype diversity was detected among the pigs at two fairs as both H1N1 and H3N2 were recovered from incoming swine. At two of the exhibitions, a temporal relationship was observed between the order of the individual swine in sampling and the associated IAV rRT‐PCR results, indicating the fomite transmission of IAV through common contact surfaces may occur. With the knowledge that a small proportion of swine arrive at fairs shedding IAV, resources should be directed towards preventive strategies focused on limiting transmission during fairs to protect swine and humans during exhibitions.     

抗流感及禽流感病毒新药"帕拉米韦"研究进展

帕拉米韦（Peramivir）是一种以流感病毒表面糖蛋白神经氨酸酶为作用靶点的新型环戊烷类抗流感药物.本文综述了帕拉米韦的合成工艺、作用机理、药效学、药动学、临床研究及毒副作用等方面的研究进展.     

新城疫、禽流感病毒同胚接种的研究

根据新城疫病毒、禽流感病毒两种病毒均能在鸡胚中繁殖的特点,将两种病毒以最适稀释比例稀释后,分别接种于9—11日龄的鸡胚尿囊腔,同时进行两种病毒同胚培养,并利用血凝试验进行病毒效价的测定。结果表明,在同一鸡胚中两种病毒均能较好地增殖,病毒间干扰现象不明显。本试验在增强疫苗的免疫原性和进一步阐明病毒的增殖机理方面具有重要意义。     

禽流感、新城疫、传染性支气管炎三联没乳剂灭活疫苗的研制

以禽流感病毒（AIV）H5N4株，新城疫病毒（NDV）LaSota株与传染性支气管炎病毒（IBV）M41株为抗原研制了AI－ND－IB三联油乳剂灭活疫苗，并对疫苗的物理性状，安全性，免疫效力，保存期及抗体消长规律进行了检测，结果表明，疫苗在免疫后3周后6个月内，对AIV－H5N4，NDV－北京株的攻击均获全部保护，在免疫后52周内，免疫鸡箅清中AIV－N5N4，MDV，IBV的HI抗体也保持较高的水平，疫苗在4℃保存15个月，其免疫力没有下降。