猪圆环病毒2型感染对猪繁殖与呼吸综合征灭活疫苗体液免疫的影响

本试验采用ELISA方法对单独接种猪繁殖与呼吸综合征病毒(PRRSV)变异株灭活苗组(HP组,n=3)和PCV2感染且出现病毒血症后接种PRRSV变异株灭活苗组(PCV2/HP组,n=3)不同时相血清中的PRRSV抗体进行检测;并对HP和PCV2/HP组血清中PCV2特异的抗体和核酸分别进行ELISA和荧光定量PCR检测。结果表明,在首免后70 d HP组血清平均抗体效价极显著高于PCV2/HP组(P<0.01);首免后56、63和77 d HP组平均抗体效价明显高于PCV2/HP组。其中在首免后56和63 d HP组抗体阳性率均达67%(2/3),PCV2/HP组在相应时相抗体阳性率为0;在首免后70和77 d HP组抗体阳性率均达100%(3/3),PCV2/HP组在相应时相抗体阳性率仅为0和33%(1/3)。结果提示PCV2感染可在一定程度上抑制PRRSV变异株(JXA1)特异性的抗体反应。     

猪圆环病毒2型感染对猪瘟弱毒疫苗免疫效果的影响

为了解猪圆环病毒2型(PCV2)感染对猪瘟(CSF)弱毒疫苗诱导的体液免疫的影响,从未接种PCV2疫苗的猪场育肥猪中选取疑似病猪和健康猪各10头,分为A、B、两组,其中A组经ELISA试剂盒检测PCV2抗体呈阳性。A、B两组同时免疫CSF弱毒疫苗,免疫后每隔2周检测CSFV抗体水平。结果显示:感染PCV2后的猪只,CSFV抗体为阴性,而未感染PCV2组在接种CSF弱毒疫苗后抗体阳性率较高,抗体水平呈现逐渐升高的趋势,提示PCV2感染对CSF弱毒疫苗的产生有明显的抑制作用。     

猪圆环病毒2型和猪瘟病毒抗体检测与分析

目的检测临汾市某县猪圆环病毒2型(PCV2)和猪瘟病毒(CSFV)的抗体水平.方法分别从该县的企业、散户、大户的猪只中采集血清186份,应用ELISA方法检测抗体水平.结果 PCV2抗体阳性率平均为83.33%,CSFV抗体阳性率平均为54.83%.结论该县猪圆环病毒2型感染严重,猪瘟疫苗免疫抗体水平偏低,与猪瘟疫苗使用不当、猪圆环病毒2型感染存在一定关系.     

猪圆环病毒2型感染对伪狂犬疫苗免疫应答的影响

<正>为了明确PCV2感染对伪狂犬(PR)疫苗免疫应答的影响,本研究彩阻断ELISA方法对单独接种猪PR疫苗组(A组)及PCV2人工感染3周后接种猪PR疫苗组(PA组)不同时相血清中的猪PR病毒gB抗体进行检测;同时对不同时相     

猪瘟兔化弱毒疫苗与猪圆环病毒2型灭活疫苗同时接种的免疫效果评价

为评价猪瘟兔化弱毒疫苗和猪圆环病毒2型(PCV2)灭活疫苗同时免疫效果,本实验采用猪瘟活疫苗(细胞源)和PCV2灭活疫苗(LG株)两种疫苗同时分点注射和混合注射35日龄仔猪,于免疫后间隔3周、5周、7周、9周采血,通过猪瘟病毒(CSFV)抗体检测试剂盒和PCV2抗体检测试剂盒进行CSFV和PCV2特异性抗体检测。结果显示,无论是两种疫苗同时颈部两侧分点分别注射,还是用PCV2灭活疫苗稀释CSFV活疫苗后混合免疫,免疫后3周CSFV和PCV2抗体均呈阳性,CSFV抗体滴度和PCV2抗体滴度于7周和9周达到峰值,与各疫苗单独免疫对照组抗体产生时间和滴度无明显差异。实验表明,两种疫苗可以同时注射或采用PCV2灭活疫苗稀释CSFV活疫苗进行免疫,均不影响各自免疫抗体的产生。因此,可以将PCV2灭活疫苗作为CSFV活疫苗的稀释剂,采用一次性注射的形式同时接种这两种疫苗,为简化这两种病临床疫苗免疫注射方面提供了实验依据。     

猪圆环病毒2型感染对伪狂犬疫苗免疫应答的影响

为明确PCV2感染对伪狂犬(PR)疫苗免疫应答的影响,本研究采用阻断ELISA方法对单独接种猪PR疫苗组(A组)及PCV2人工感染3周后接种猪PR疫苗组(PA组)不同时相血清中的猪PR病毒gB抗体进行检测;同时对不同时相前腔静脉血进行CD4+/CD8+流式细胞术及血常规分析。结果表明,在PCV2感染后2周至5周间,A组白细胞含量均高于PA组,随后PA组白细胞恢复至与A组略高的正常水平;在整个实验中,除接种猪PR疫苗后1周(WPI)和9周(WPI)外的所有时相PA组的淋巴细胞含量均略高于A组;PCV2感染后可使记忆/激活Th细胞数量略有升高,幼稚型Th细胞含量的下降;PCV2感染后2周～7周PA组Tc细胞均高于A组,在9WPIPA组Tc细胞数量显著下降(p0.05);除9WPI外,A组的S/N值均低于PA组。结果表明,PCV2感染看降低机体产生针对PRVgB特异性抗体水平,而且在一定程度上降低了幼稚型Th细胞及Tc细胞含量。     

猪圆环病毒2型感染对猪瘟弱毒疫苗接种猪免疫应答的影响

为探索猪圆环病毒2型(PCV2)感染对猪瘟(CSF)弱毒疫苗接种猪免疫应答的影响,将20头28 d断奶仔猪随机分为V-I、I-V、V和C4组,5头·组-1.V-I组在接种CSF弱毒疫苗后2d感染PCV2;I-V组在感染PCV2 后2d接种CSF弱毒疫苗;V组只接种CSF弱毒疫苗;C组为空白对照组.共免疫2次,间隔21 d.免疫后定期检测血清CSFV特异性抗体水平、外周血淋巴细胞(PBLC)增殖活性和PBLC中IFN-γ、IL-2、IL-4和IL-10 mRNA的表达水平.结果显示:在CSF弱毒疫苗免疫前或免疫后感染PCV2,均会导致CSFV抗体水平低下,血清抗体阳转率下降,PBLC增殖活性降低.初次免疫后,V-I与I-V组的PBLC内IFN-γ、IL-2、IL-4和IL-10 mRNA的表达量严重不足,其中V-I组的表达量最低.加强免疫后V-I与I-V组的PBLC内IFN-γ与IL-10的表达失衡,IL-2和IL-4的表达缺乏,其中I-V组的细胞因子表达失衡和缺乏更严重.上述研究表明,CSF弱毒疫苗免疫前或免疫后感染PCV2均会影响机体的体液和细胞免疫应答水平,导致PBLC内细胞因子的表达严重抑制和紊乱.     

小单元全封闭隔离栏舍在猪疫病控制与净化上的效果研究

【目的】 研究小单元全封闭隔离栏舍在猪疫病控制、净化及疫苗减免上的应用效果。【方法】 试验设置猪圆环病毒2型（Porcine circovirus type 2,PCV2）血清抗体全阴、全阳及部分阳性组,每组选取同日龄的保育猪6头,进行为期140 d的A、B模式饲养。A模式为小单元全封闭隔离栏舍养殖140 d,B模式为小单元全封闭隔离栏舍养殖70 d＋常规敞开型栏舍养殖70 d。所有试验猪进行猪瘟病毒（Classical swine fever virus,CSFV）和猪伪狂犬病病毒（Pseudorabies virus,PRV）单次疫苗免疫。试验在饲养的第0、70和140天检测猪群血清PCV2、猪繁殖与呼吸综合征病毒（Porcine reproductive and respiratory syndrome virus,PRRSV）、CSFV、PRV-gB、PRV-gE抗体水平及血清抗体阳性率。【结果】 与第0天相比,A模式所有组PCV2抗体水平及全阳组和部分阳性组PCV2血清抗体阳性率在第70和140天显著下降（P<0.05）,B模式第70天全阳组PCV2抗体水平极显著下降（P<0.01）,全阴组及部分阳性组PCV2抗体水平维持,第140天所有组PCV2抗体水平均极显著升高（P<0.01）。B模式PCV2血清抗体阳性率前70 d大幅度下降,后70 d大幅度上升至100%。与第0天相比,A、B模式前70 d CSFV抗体水平显著升高（P<0.05）,血清抗体阳性率快速升高;与第70天相比,后70 d A模式全阴组和部分阳性组CSFV抗体水平维持小幅度升高趋势,全阴组和全阳组血清抗体阳性率维持100%,B模式全阴组和全阳组CSFV抗体水平显著下降（P<0.05）并接近第0天。PRV-gB血清抗体水平及阳性率在A、B两种养殖模式的前70 d部分组下降,后70 d全部组下降。在整个试验期内A、B养殖模式下,PRRSV和PRV-gE抗体水平均处在较低水平,血清抗体阳性率均为0。【结论】 小单元全封闭隔离栏舍可明显阻断PCV2的传播,具有净化PCV2、增强CSFV疫苗免疫的保护率、降低疫苗使用成本的作用,是一种可行的增强生猪养殖疾病防御能力的保障设备。     

猪圆环病毒2型感染对口蹄疫O型合成肽疫苗免疫应答的影响

采用阻断ELISA方法对单独接种口蹄疫O型(FMD)组(C组,n=3)及PCV2人工感染4周后接种FMDO型疫苗组(PC组,n=4)后不同时相血清中的FMDO型抗体进行检测;同时对不同时相前腔静脉血进行CD4/CD8流式细胞术及血常规分析。结果显示,除接种后4周C组白细胞及淋巴细胞含量与略低于PC组外,接种疫苗后2～8周C组白细胞及淋巴细胞含量均略高于CP组,且在接种疫苗后2周,C组淋巴细胞含量显著高于PC组(P0.05);在整个试验过程中PC组的幼稚型Th细胞含量均高于C组,且在接种疫苗后0周和8周2组间幼稚型Th细胞含量差异显著(P0.05);在接种疫苗后2～6周期间,除在接种疫苗后4周2组间Tc细胞含量较为接近外,C组的Tc细胞含量均高于PC组;除在8WPIPC组CD4+CD8+记忆/激活Th细胞含量略高于C组外,在接种疫苗后0～6周过程中,C组记忆/激活Th细胞含量均略高于PC组,且在接种疫苗后6周,2组间记忆/激活Th细胞含量差异显著(P0.05);在接种前疫苗4周和前2周(-4-2WPI)期间,C组的抗体阻断率均高于PC组,在接种疫苗后4～8周2组间抗体阻断率差异逐渐变小,且PC组抗体阻断率略高于C组。结果表明,PCV2感染对机体产生针对FMDO型特异性抗体早期有一定的抑制作用,但在后期这种抑制作用逐步减弱,甚至消失;PCV2感染确实可以在一定程度上抑制机体对猪FMDO型合成肽疫苗体液免疫应答,导致机体的淋巴细胞及白细胞含量降低,进而导致Tc细胞及记忆/激活Th细胞含量下降。     

某猪场4种猪病毒性疫病抗体水平检测及分析

为了解贵州省某规模化猪场4种猪常见病毒性疫病的抗体水平及感染情况,实验采用ELISA抗体检测法,对该猪场送检的370份血清进行了CSFV、PRRSV、PCV2和PRV 4种抗体水平检测。实验结果表明:该猪场种猪CSFV、PRRSV、PCV2和PRV抗体阳性率分别为100%、94.74%、100%、5.26%,保育猪CSFV、PRRSV、PCV2和PRV抗体阳性率分别为94.44%、83.33%、16.7%、5.56%;其中种猪CSF、PRRS、PCV2、PR已接种疫苗;保育猪CSF、PRRS、PR已接种疫苗,未接种PCV2疫苗,上述数据说明该猪场种猪CSF、PCV2、PRRS、PR免疫效果较好;保育猪CSF、PRRS、PR免疫效果较好,PCV2有野毒感染现象。该猪场种猪应在保育猪阶段做好PCV2的预防接种工作,同时做好猪场的疫病净化措施。     

Porcine circovirus type 2 (PCV2) infection decreases the efficacy of an attenuated classical swine fever virus (CSFV) vaccine

ABSTRACT: The Lapinized Philippines Coronel (LPC) vaccine, an attenuated strain of classical swine fever virus (CSFV), is an important tool for the prevention and control of CSFV infection and is widely and routinely used in most CSF endemic areas, including Taiwan. The aim of this study was to investigate whether PCV2 infection affects the efficacy of the LPC vaccine. Eighteen 6-week-old, cesarean-derived and colostrum-deprived (CDCD), crossbred pigs were randomly assigned to four groups. A total of 105.3 TCID50 of PCV2 was experimentally inoculated into pigs through both intranasal and intramuscular routes at 0 days post-inoculation (dpi) followed by LPC vaccination 12 days later. All the animals were challenged with wild-type CSFV (ALD stain) at 27 dpi and euthanized at 45 dpi. Following CSFV challenge, the LPC-vaccinated pigs pre-inoculated with PCV2 showed transient fever, viremia, and viral shedding in the saliva and feces. The number of IgM+, CD4+CD8-CD25+, CD4+CD8+CD25+, and CD4-CD8+CD25+ lymphocyte subsets and the level of neutralizing antibodies against CSFV were significantly higher in the animals with LPC vaccination alone than in the pigs with PCV2 inoculation/LPC vaccination. In addition, PCV2-derived inhibition of the CSFV-specific cell proliferative response of peripheral blood mononuclear cells (PBMCs) was demonstrated in an ex vivo experiment. These findings indicate that PCV2 infection decreases the efficacy of the LPC vaccine. This PCV2-derived interference may not only allow the invasion of wild-type CSFV in pig farms but also increases the difficulty of CSF prevention and control in CSF endemic areas.     

表达猪瘟病毒E2蛋白的重组腺病毒对猪的免疫效力评价

为了进一步验证含有猪瘟病毒E2基因的重组腺病毒（rAdV—E2）在猪体上的免疫效力,将rAdV—E2按108TCID50／头接种猪2次,同时用野生型腺病毒wtAdV作为阴性对照,当抗体上升到一定程度后用致死剂量的猪瘟强毒石门株进行攻击。结果表明,rAdV—E2免疫组（n=5）所有免疫猪在加强免疫后均产生了猪瘟特异性中和抗体,并于加强免疫后3w达到峰值,攻毒后所有猪只抗体迅速升高,除了一头猪短期体温升高外,未出现任何其它临床症状;而野生型腺病毒wtAdV免疫组（n=5）猪只在攻毒前一直没有检出特异性抗体,攻毒后全部出现典型的猪瘟临床症状和严重的病毒血症,剖检时可见典型猪瘟病理变化。这表明构建的猪瘟病毒E2基因重组腺病毒rAdV—E2免疫猪后产生了很好的免疫效果,有望成为具有开发价值的活载体疫苗。     

河南省部分猪群主要疫病疫苗免疫抗体水平监测与效果分析

2009年1月—2011年12月,从河南省信阳和驻马店地区62个猪场共收集免疫过猪瘟病毒(CSFV)、伪狂犬病毒(PRV)、猪繁殖与呼吸综合征病毒(PRRSV)和猪圆环病毒2型(PCV2)疫苗的猪血液,利用ELISA方法对样品进行抗体水平的检测。结果显示:该地区猪场猪群的疫苗免疫合格率最高是PRV疫苗(87.06%);其次是CSFV疫苗(76.24%);PRRSV和PCV2的疫苗免疫合格率较低,分别为65.20%和50.78%。4种疫苗免疫抗体合格率在不同规模的猪场有较大的差别,规模猪场的猪群某些疫病的抗体未必比散养猪群高。种猪群的4种疫苗的免疫合格率最高,商品猪中各个生长阶段的免疫后抗体合格率比较后发现,断奶仔猪群PRRSV抗体合格率明显高于哺乳仔猪,育肥猪群合格率要比哺乳仔猪群和断奶仔猪群的抗体合格率低。     

Efficacy of a live attenuated vaccine in classical swine fever virus postnatally persistently infected pigs

Classical swine fever (CSF) causes major losses in pig farming, with various degrees of disease severity. Efficient live attenuated vaccines against classical swine fever virus (CSFV) are used routinely in endemic countries. However, despite intensive vaccination programs in these areas for more than 20 years, CSF has not been eradicated. Molecular epidemiology studies in these regions suggests that the virus circulating in the field has evolved under the positive selection pressure exerted by the immune response to the vaccine, leading to new attenuated viral variants. Recent work by our group demonstrated that a high proportion of persistently infected piglets can be generated by early postnatal infection with low and moderately virulent CSFV strains. Here, we studied the immune response to a hog cholera lapinised virus vaccine (HCLV), C-strain, in six-week-old persistently infected pigs following post-natal infection. CSFV-negative pigs were vaccinated as controls. The humoral and interferon gamma responses as well as the CSFV RNA loads were monitored for 21 days post-vaccination. No vaccine viral RNA was detected in the serum samples and tonsils from CSFV postnatally persistently infected pigs for 21 days post-vaccination. Furthermore, no E2-specific antibody response or neutralising antibody titres were shown in CSFV persistently infected vaccinated animals. Likewise, no of IFN-gamma producing cell response against CSFV or PHA was observed. To our knowledge, this is the first report demonstrating the absence of a response to vaccination in CSFV persistently infected pigs.     

猪瘟疫苗在猪圆环病毒2型阳性猪场的免疫效果观察

为了研究猪圆环病毒2型(PCV-2)感染对猪瘟疫苗免疫效力的影响,对PCR证实为PCV-2阳性的试验猪进行猪瘟疫苗的免疫,分别在免疫后第1、3、7、14、21、28和63天对试验猪进行猪瘟病毒(CSFV)和PCV-2的抗体检测。检测结果表明PCV-2阳性猪在猪瘟疫苗免疫后均未能产生有效的CSFV抗体,从免疫猪的血液和内脏组织中也未能检测到CSFV核酸。虽然只能从1头PCV-2阳性猪的血清中检测到PCV-2抗体,但是却能从全部实验猪的淋巴结中检测到PCV-2的核酸。试验猪的病理组织学观察和白细胞计数也表明PCV-2阳性猪的淋巴结呈典型的PCV-2感染的病理变化,且白细胞数量显著低于健康猪。表明PCV-2的感染会对猪的免疫系统造成损害从而抑制猪瘟疫苗的免疫效果。     

Yeast-expressed classical swine fever virus glycoprotein E2 induces a protective immune response

Classical swine fever (CSF) is an economically important swine disease worldwide. The glycoprotein E2 of classical swine fever virus (CSFV) is a viral antigen that can induce a protective immune response against CSF. A recombinant E2 protein was constructed using the yeast Pichia pastoris expression system and evaluated for its vaccine efficacy. The yeast-expressed E2 (yE2) was shown to have N-linked glycosylation and to form homodimer molecules. Four 6-week-old specified-pathogen-free (SPF) piglets were intramuscularly immunized with yE2 twice at 3-week intervals. All yE2-vaccinated pigs could mount an anamnestic response after booster vaccination with neutralizing antibody titers ranging from 1:96 to 1:768. Neutralizing antibody titers at 10 weeks post booster vaccination ranged from 1:16 to 1:64. At this time, the pigs were subjected to challenge infection with a dose of 1 × 10⁵ TCID₅₀ (50% tissue culture infective dose) virulent CSFV strain. At 1 week post challenge infection, all of the yE2-immunized pigs were alive and without symptoms or signs of CSF. Neutralizing antibody titers at this time ranged from 1:4,800 to 1:12,800 and even to 1:51,200 one week later. In contrast, the control pigs continuously exhibited signs of CSF and had to be euthanized because of severe clinical symptoms at 6 days post challenge infection. All of the yE2-vaccinated pigs were E^rns antibody negative and had seroconverted against E^rns by post challenge day 11, suggesting that yE2 is a potential DIVA (differentiating infected from vaccinated animals) vaccine. The yeast-expressed E2 protein retains correct immunogenicity and is able to induce a protective immune response against CSFV infection.     

猪瘟野毒感染抗体阻断ELISA检测方法的建立与初步评价

以猪瘟野毒E2蛋白为包被抗原、辣根过氧化物酶标记的猪瘟野毒单抗作为酶标抗体,建立检测猪瘟野毒抗体的阻断ELISA方法。猪瘟野毒E2最适包被浓度为0.03μg/mL,待检血清最适稀释度为1∶4,酶标猪瘟野毒单抗稀释度为1∶1 000。用建立的阻断ELISA方法检测369份临床阴性血清,计算阻断率,确定临界值,阻断率>40%为猪瘟野毒抗体阳性,阻断率≤40%为猪瘟野毒抗体阴性。用建立的ELISA方法检测84份血清,其中78份为免疫猪瘟疫苗的血清,6份为猪瘟病毒感染血清。结果显示,78份免疫血清均检测为猪瘟野毒抗体阴性,6份猪瘟感染血清均检测为猪瘟野毒抗体阳性。因此可初步判定该方法可用于鉴别诊断猪瘟病毒自然感染动物和C株疫苗免疫动物的血清抗体,并为临床检测猪瘟野毒抗体提供便捷、快速,精准的检测工具,对猪瘟的临床诊断、预防以及猪瘟净化工作具有非常重要的参考意义。     

2017-2020年浙江省猪主要病毒性传染病的流行病学调查与分析

为了解浙江省猪主要病毒性传染病的流行情况及变化规律,本试验对2017-2020年浙江省不同地区规模化猪场送检的血清和病料样品进行猪繁殖与呼吸综合征病毒(PRRSV)、猪瘟病毒(CSFV)、猪圆环病毒2型(PCV2)和猪伪狂犬病病毒(PRV)4种疫病痛原和抗体检测.抗体检测结果显示:PRRSV、CSFV、PCV2、PRV...