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.     

定量检测马动脉炎病毒方法的建立

为建立快速、敏感、准确的马动脉炎病毒检测方法,笔者选取病毒基因组中高度保守的ORF7序列设计引物和TaqMan探针,分别使用马动脉炎病毒的总RNA和含有选定检测序列的克隆标准品进行一步法实时定量RT-PCR,绘制扩增曲线,两曲线斜率之差小于0.1,证明两者的扩增效率相同,可用选定检测序列的克隆标准品对马动脉炎病毒进行定量检测。     

鸡病毒性关节炎活疫苗对 SPF 鸡攻毒保护实验研究

本研究分别测定了两种病毒性关节炎活疫苗ZJS株和S1133株的TCID50。其中ZJS疫苗株的TCID50为104.5TCID50/羽份,S1133疫苗株的TCID50为103.5TCID50/羽份。通过进一步的攻毒保护实验,证实ZJS疫苗株对SPF鸡的临床保护力明显优于S1133疫苗株。     

牛病毒性腹泻病毒弱毒活疫苗免疫持续期的研究

为检测牛病毒性腹泻病毒(BVDV)弱毒活疫苗在免疫牛体内抗体产生及其消长规律,评价弱毒疫苗的保护效力,并确定免疫持续期,本试验对免疫试验牛每头颈部肌肉接种BVDV SM株弱毒疫苗104.5TCID50,监测血清抗体效价,进行免疫持续期的确定。在疫苗免疫后的6个月、9个月和12个月,分别抽取5头免疫组和5头对照组牛,采用BVDV-JL强毒株进行攻毒试验,每头牛攻毒剂量为6×107.0TCID50/mL。结果显示疫苗免疫后12个月时血清中和抗体效价仍维持在1∶1048以上。攻毒结果显示,在3个不同时间点进行强毒攻击后,免疫组所有动物白细胞数量都没有下降也没有分离到病毒,而对照组动物白细胞数下降均超过30%,6个月和9个月时动物血清中均能分离到病毒,而12个月对照组动物由于年龄大,没有分离到病毒,因此暂定此疫苗的免疫持续期为9个月。     

牛病毒性腹泻弱毒活疫苗免疫持续期的研究

为检测牛病毒性腹泻病毒(BVDV)弱毒活疫苗在免疫牛体内抗体产生及其消长规律,评价弱毒疫苗的保护效力,并确定免疫持续期,本试验对免疫试验牛每头颈部肌肉接种BVDV SM株弱毒疫苗10^4.5TCID₅₀/头,监测血清抗体效价,进行免疫持续期的确定。在疫苗免疫后的6、9和12个月分别抽取5头免疫组和5头对照组牛采用BVDV-JL强毒株进行攻毒试验,每头牛攻毒剂量为6×10^7.0 TCID₅₀/mL。结果显示疫苗免疫后12个月时血清中和抗体效价仍维持在1∶1048以上,攻毒结果显示3个时间点强毒攻击后,免疫组所有动物白细胞数量都没有下降也没有分离到病毒,而对照组动物白细胞数下降均超过30%,6和9个月动物均分离到病毒,而12个月对照组动物由于年龄大,没有分离到病毒,因此暂定此疫苗的免疫持续期为9个月。     

Interstate Equine Semen and Embryo Shipment Regulations in the United States and Their Implications on Control of Disease Transmission

Outbreaks of both equine viral arteritis and contagious equine metritis (CEM) in the United States have occurred in recent years. Shipped semen has been implicated in disease transmission and rapid spread. A survey was performed of state regulations regarding testing for equine arteritis virus (EAV) and CEM before interstate semen or embryo shipment. Results demonstrated lack of any requirements in 31 states. Four states had regulations regarding EAV; 17 required a Certificate of Veterinary Inspection (CVI) to accompany a semen or embryo shipment. Ten states required a negative equine infectious anemia test, primarily as a requirement of a CVI versus because of risk of dissemination of disease. No states required CEM testing. A comparison of state department of agriculture or veterinary medical association Web sites and direct communication with state veterinarians or their offices demonstrated contradictory information in six states. The lack of uniform regulations concerning CEM and EAV testing for shipped semen and embryos, and the threat they pose to the equine breeding industry and horse health, should be alarming to veterinarians and horse owners. Routine testing of animals before shipment of semen or embryos and veterinary involvement in collection and breeding activities are appropriate and necessary to help prevent future outbreaks and protect equine health.     

犬瘟热弱毒疫苗免疫犬抗体消长规律及影响因素

应用犬瘟热病毒（ＣＤＶ）血清中和试验，对从日本引进的犬瘟热弱毒疫苗免疫犬的血清中和抗体效价进行了检测，探讨了免疫剂量、免疫程序和犬的年龄对抗体效价的影响。结果表明，试验犬于第１次免疫后１４ｄ左右产生１∶９以上的血清中和抗体，随后抗体水平迅速升高，３５ｄ时７８．３８％的免疫犬抗体效价超过１∶１００，６０ｄ达到高峰，６个月试验结束时，血清中和抗体效价仍维持在１∶１００以上。该犬瘟热弱毒疫苗的最低免疫剂量为１０３．０ＴＣＩＤ５０，以２周或３周间隔连续免疫３次均可使免疫犬获得完全保护。     

Experimental Infection of Calves with Bovine Viral Diarrhoea Virus Type-2 (BVDV-2) Isolated from a Contaminated Vaccine

A non-cytopathic strain of BVDV-2 was isolated from a batch of live infectious bovine rhinotracheitis (IBR) vaccine, and inoculated intranasally into four 3-month-old calves. Severe signs of disease developed by days 4 and 6 in three of the calves, free of BVDV and antibodies to BVDV, that had been exposed to the virus. These calves survived the acute phase of the infection and progressively recovered. BVDV was consistently isolated, or the respective viral RNA was detected, in the buffy coats from blood samples collected starting from days 2 or 4 up to days 11 or 14 after the experimental infection. Viral RNA was also detected in sera from these infected calves until the presence in the serum of virus neutralizing antibodies was demonstrated. By contrast, the only calf having pre-existing neutralizing antibodies to BVDV at the start of the study was protected from the disease. No virus was detected at any time after experimental inoculation of this calf. Genomic characterization of the BVDV-2 isolated in cell cultures, or detected in sera from the experimentally infected animals, revealed 100% homology in the nucleotide sequence with the BVDV-2 detected as a contaminant of the live IBR virus vaccine. These findings provided evidence of the infective nature of the contaminant BVDV-2 and of its potential to generate disease outbreaks when inoculated into susceptible animals.     

Equine viral arteritis: A respiratory and reproductive disease of significant economic importance to the equine industry

Equine arteritis virus is the causative agent of equine viral arteritis, a respiratory and reproductive disease that affects the members of the family Equidae. The virus was first isolated from the lung of an aborted fetus after an extensive outbreak of respiratory disease and abortion on a Standardbred breeding farm near Bucyrus, Ohio, in 1953. Since then, periodic outbreaks of equine viral arteritis have been reported in a number of countries around the world. This disease may result in significant economic loss to the equine industry due to the occurrence of abortion in pregnant mares, neonatal mortality, and establishment of the carrier state in stallions. This article provides an extensive review on equine arteritis virus, epidemiology, disease, pathogenesis, and prevention and control measures.     

马动脉炎病毒N蛋白基因GST融合表达载体的构建及表达

采用RT-PCR方法扩增出马动脉炎病毒核衣壳蛋白基因0RF7,并将其克隆到pMDl8-T载体,构建成重组质粒pMDl8-N,在上海生物工程公司测序。结果表明,所克隆的核衣壳蛋白基因序列与EAVNC-002532株的同源性为99％。表明0RF7是EAV基因组内的保守序列,将0RF7亚克隆到原核表达载体pGEX-6P-1中,构建成重组质粒pGEX-6P-N,用pGEX-6P-N转化表达菌株BL21（DE3）,诱导表达后SDS-PAGE和Westernblotting分析表明,克隆在谷胱苷肽硫转移酶（GST）下游的核衣壳蛋白基因与GST获得了高效融合表达,表达的融合蛋白GST-N分子质量约为40ku,为马动脉炎病毒病血清学诊断方法的建立奠定了基础。     

Recombinant Strain A/HK/Otar/6:2/2010 (H3N8) for Development of a Live Intranasal Equine Influenza Vaccine

Recombinant cold-adapted strain A/HK/Otar/6:2/2010 (H3N8) constructed by the method of classic genetic reassortment, inherited the hemagglutinin and neuraminidase genes from the epizootic A/equine/Otar/764/2007 (Н3N8) virus and genes of nonglycosylated proteins from attenuation donor A/Hong Kong/1/68/162/35 (H3N2). The resulted virus was of cold-adapted (efficient growth at 25°C) and temperature-sensitive (restricted replication at 39°C) phenotype. Like the parental virus of subtype H3N2, the resulted reassortant was attenuated, with limited replication in lungs of mice (1.75 lgEID₅₀/mL) versus replication in turbinate (2.5 lgEID₅₀/mL). Intranasal immunization of mice with A/HK/Otar/6:2/2010 (H3N8) reassortant did not induce animal weight reduction in contrast to epizootic A/equine/Otar/764/2007 (Н3N8) virus.     

布鲁氏菌病弱毒活疫苗经不同黏膜途径免疫奶牛的排菌程度及免疫反应

为探究布鲁氏菌病疫苗经不同黏膜途径免疫奶牛后对体液免疫与细胞免疫水平及疫苗体外排菌方面的影响,将80头10~12月龄的奶牛平均分为四组,通过眼部滴注(A组)、口腔喷注(B组)、阴道灌注(C组)三种黏膜免疫途径对奶牛接种布鲁氏菌病弱毒活疫苗(A19株),并与皮下注射免疫组(D组)进行比较,测定了免疫后7 d内黏膜免疫组的排菌情况,180 d内四组免疫奶牛的抗体效价和60 d内四组免疫奶牛的外周血淋巴细胞细胞因子分泌(IFN-γ、IL-2和IL-4)水平。结果显示:黏膜免疫组在免疫后5 d内均有不同程度的排菌,以免疫后1 d内排菌量为最高,并且在黏膜免疫组中阴道灌注组排菌量最低;各组奶牛在免疫后20 d时MSAT测定抗体效价平均值均上升至最高,之后持续下降,黏膜免疫组的抗体水平显著低于注射免疫组,且在黏膜免疫组中阴道灌注组的抗体水平最高;A组和B组奶牛在免疫后120 d RBT检测无阳性,C组在免疫后150 d RBT检测无阳性,D组在免疫后180 d RBT检测阳性率35%;B组、C组在免疫后7 d,A组、D组在免疫后20 d IFN-γ分泌量达到最高值,各组IL-2和IL-4分泌量未见明显变化,黏膜免疫组中阴道免疫组的IFN-γ分泌量为最高,仅次于注射免疫。结果表明,阴道灌注是一种较优的奶牛布鲁氏菌病黏膜免疫方式。     

犬细小病毒基因疫苗对幼貉的免疫试验

应用已构建的pcDCPV和pIRCPVIL2重组质粒配制成疫苗,对幼貉进行免疫,并对其免疫反应进行观察。应用夹心ELISA方法检测外周血中特异性的抗细小病毒血清抗体免疫球蛋白(IgG)的水平,MTT比色法检测外周血中淋巴细胞的转化率,对基因疫苗的免疫保护进行观察。结果表明,用pcDCPV和pIRCPVIL2重组质粒免疫后的幼貉对细小病毒的攻击具有一定的保护性,为pcDCPV和pIRCPVIL2重组质粒的应用奠定了基础。     

马传贫弱毒疫苗接种马(骡)免疫反应动力学研究

本试验对注射马传贫弱毒疫苗后的马(骡)50匹,分别进行了体温、血液学、血清抗体及血液中T淋巴细胞等项目的检测。在此基础上,探讨了注苗后马(骡)免疫反应及其动态规律。提出注苗马(骡)在T细胞恢复到正常水平前,仍有感染强毒的可能,因此对马(骡)仍需进行监控。     

牛流行热病毒灭活疫苗免疫后牛外周血淋巴细胞亚类分布的研究

对牛流行热病毒灭活疫苗免疫并攻每后，牛外周血淋巴细胞亚类的变化进行了研究。结果表明，灭活苗免疫后，牛的CD4^ 显著升高，可能与参与辅助B淋巴细胞合成抗体有关，攻毒后，γδT细胞均显著上升，免疫组在攻毒后3周仍保持在相当的高水平，IL－2Rα阳性细胞在攻毒后高热期也有显著升高，而CD8^ 细胞没有明显变化。     

A study of the effects of Marek's disease in a broiler flock

In a flock of 17000 broilers, characteristic Marek's disease (MD) paralysis, accompanied by increased mortality occurred during the fourth week of the flock's life. Cytolytic lesions and atrophy in the bursae of Fabricius, and the severity of lymphocytic infiltration of nerves, livers, proventriculus and brain are suggestive of the involvement of MD virus of greater pathogenicity than has been encountered previously in New Zealand.     

Vaccination Response of Young Foals to Keyhole Limpet Hemocyanin: Evidence of Effective Priming in the Presence of Maternal Antibodies

The ability to produce antibodies is essential for protection from infectious disease; however, in the neonate, maternal antibodies have been proposed to interfere with the foal's ability to respond to vaccination. In species other than the equid, keyhole limpet hemocyanin, a high-molecular weight protein, is used in vivo as an experimental vaccine component because of its high intrinsic immunogenicity. In this study, we show that young foals are able to produce a primary antibody response to vaccination at an early age. Thus, foals, like human infants, are capable of responding to antigenic exposure to a novel antigen (keyhole limpet hemocyanin) during the neonatal period. Although vaccinating foals in the presence of maternal antibodies failed to induce a primary serological response, priming occurred as comparable anamnestic responses were detected upon subsequent exposure to the antigen. There was no evidence of tolerance induction.     

Treatment of Metastatic Equine Melanoma with a Plasmid DNA Vaccine Encoding Streptococcus Pyogenes EMM55 Protein

A 19-year-old castrated male Arab/Quarter horse presented with an extensive history of cutaneous metastatic melanoma. Over a period of 8 months, a total of 8 doses of plasmid DNA vaccine expressing the Streptococcus pyogenes emm55 gene (pAc/emm55) were administered intratumorally at 300 μg/dose via a needless injector. Upon completion of the vaccination protocol, the size of the injected lesions, on average, were reduced by 40.3% from the initial size measurements. Lesions that were not injected were reduced by 47.6%. The overall reduction in total tumor burden was 42.3%. Tumor regression was also associated with the augmentation of antimelanoma IgG antibody response, thus implying that an induction of an effective antimelanoma response would be of great advantage in the management of equine melanoma.     

牛病毒性腹泻-牛传染性鼻气管炎多价二联蜂胶灭活苗的研究

牛病毒性腹泻、牛传染性鼻气管炎是牛的主要传染病,采用我所分离的地方毒株,以蜂胶为免疫佐剂制成多价二联灭活苗,经免疫及攻毒试验、二联灭活苗保护率的中间试验等实验,表明该苗对牛的保护性良好。