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1.
Groups of two or three day old pigs were inoculated intravenously with cell culture grown transmissible gastroenteritis virus. A single or a multiple dosage schedule was used. The magnitude of immune response was measured in terms of serum neutralization indices. A single dose of relatively attenuated virus caused mild clinical signs of transmissible gastroenteritis infection in the pigs and induced a low level of antibody in the serum by the seventh day after inoculation. Repeated injections of virus at seven day intervals stimulated little increase in antibody titers. However, high serum antibody titers were obtained for all pigs if the time interval between injections was extended to 15 days. Sera obtained early after exposure to live transmissible gastroenteritis virus contained mainly IgM antibody whereas sera obtained later after exposure contained mainly IgG antibody. Ten plaque purified isolates of transmissible gastroenteritis virus, comprising eight American isolates, one Japanese isolate and one British isolate were indistinguishable by means of reciprocal plaque reduction neutralization tests.  相似文献   

2.
Summary

Direct diagnosis of swine influenza infection by an indirect immunofluorescence technique using anti‐nucleoproteine monoclonal antibody was compared with virus isolation. Five 8‐week‐old pigs were inoculated with 2 × 107 EID50 of strain A H1N1 Sww//4115/85. Clinical signs developed in only three pigs. Antigen was detected in nasal epithelial cells obtained from all animals the first day after inoculation; the antigen was detected in one pig 6 days after the infection. Fluorescence was present in the nucleus, nucleolus and cytoplasm of infected cells. The indirect immunofluorescence test was specific and as sensitive as virus isolation in embryonated eggs, allowing a rapid diagnosis that could be achieved within hours.  相似文献   

3.
Thirty-three pigs in three groups of nineteen, ten, and four pigs were infected with three different African swine fever (ASF) virus isolates, respectively. All virus isolates were attenuated to varying degrees by passaging in cell cultures, and they retained sufficiently low virulence to produce subacute and chronic infections in pigs. Sera collected at various intervals were tested for antibody activity by the immunoelectroosmophoresis, agar gel diffusion precipitin, and complement-fixation tests using a modified Kolmer technique. Results clearly indicated that the immunoelectroosmophoresis test is a rapid (30 minute) and accurate method with extreme sensitivity and superior to the complement-fixation and agar gel diffusion precipitin tests in detecting antibody against ASF virus. Possible use of this method in detecting ASF virus infection is suggested.  相似文献   

4.
An immunohistochemical study of the tonsils was carried out to gain further insight in the pathogenesis of acute African swine fever (ASF). Twenty-one pigs were inoculated by intramuscular route with a highly virulent isolate of ASF virus and painlessly killed at 1-7dpi. Viral antigen was highly distributed in the tonsil from 3 to 4dpi and an increase in the number of monocyte-macrophages was very evident at the same days post inoculation. This phenomenon was observed together with an increase of the expression of proinflammatory cytokines (Tumour necrosis factor alpha and Interleukin-1 alpha) and the apoptosis of lymphocytes studied by the terminal deoxynucleotidyltransferase-mediated dUTP nick end labelling (TUNEL) technique and haemorrhages. With these results, we can conclude that the tonsil is suffering similar lesions than those observed in other lymphoid organs in acute African swine fever, even when the route of inoculation is the intramuscular and not oral-nasal.  相似文献   

5.
非洲猪瘟(African swine fever,ASF)是由非洲猪瘟病毒(African swine fever virus,ASFV)引起的家猪和野猪的急性、出血性传染病,强毒株感染猪的致死率接近100%,给养猪业造成巨大经济损失。ASFV在猪群中可以快速有效传播,在环境中可稳定存在,为ASF的防控带来了挑战。研究人员一直致力于ASF疫苗的研究,迄今为止,仍没有有效的疫苗投入市场。综述了ASF灭活疫苗、减毒活疫苗、亚单位疫苗、DNA疫苗、病毒活载体疫苗的研究情况,以期为ASF疫苗的有效研发提供参考。  相似文献   

6.
Mechanism of thrombocytopenia in African swine fever   总被引:1,自引:0,他引:1  
Pigs were inoculated with an African swine fever (ASF) isolate of moderate virulence, and the changes in the number of circulating blood platelets during infection were correlated with the appearance of antiviral antibody and fluctuations in total plasma hemolytic complement concentrations. Thrombocytopenia was detected by postinoculation days (PID) 7 and 8, and antiviral antibody was detected by PID 7, using an indirect immunofluorescence technique. The total hemolytic complement concentration was moderately and transiently decreased from PID 5 to 9, but was consistently low from PID 18 to 26. Pigs inoculated with an ASF virus isolate of greater virulence had a decrease in platelet counts on PID 6 and 7, and the total plasma hemolytic complement levels decreased in all pigs by PID 6 to 7. Antibody to ASF virus was not detected in pigs inoculated with the more virulent isolate. Pigs sensitized to ASF viral antigen with an inactivated-virus vaccine or by previous infection with ASF were challenge exposed. Sensitized pigs became clinically ill and thrombocytopenic by 24 to 72 hours earlier than did inoculated, nonsensitized pigs. Vaccinated pigs inoculated with homologous virus had lower blood virus concentrations than did nonvaccinated pigs. African swine fever virus-sensitized pigs inoculated with heterologous virus had a higher fatality rate than did nonsensitized pigs, and the pigs died peracutely, with only a few gross lesions in evidence. In vitro experiments demonstrated that ASF virus antigen induced platelet aggregation in platelet-rich plasma from recovered, nonviremic pigs. Viral antigen, antibody, or complement was not demonstrable on the surface of platelets from pigs inoculated with ASF virus isolate, by direct immunofluorescence testing.  相似文献   

7.
Coronavirus-like particles were visualized by electron microscopy in the intestinal contents of piglets during a diarrheal outbreak on a Quebec pig farm. The precipitating antigens of transmissible gastroenteritis virus were not detected in the intestinal contents of diarrheic animals by counter-immunoelectrophoresis. Insignificant antibody titers against transmissible gastroenteritis virus were demonstrated in the sera of convalescent pigs by indirect immunofluorescence and these sera did not react with transmissible gastroenteritis virus when tested by immunoelectron microscopy. The causative agent could not be isolated in cell cultures. It was concluded that a coronavirus antigenically distinct from transmissible gastroenteritis virus was responsible for the enteric problems observed on this farm. The outbreak was controlled after oral inoculation of adult pigs with infected intestinal contents.  相似文献   

8.
V. Cultivation of the Virus in Primary Pig Kidney Cells   总被引:3,自引:1,他引:2       下载免费PDF全文
Six strains of African swine fever (ASF) virus were propagated in culture of primary pig kidney (PK) cells. The course of virus growth was followed by means of the fluorescent antibody staining technique. All 6 strains multiplied in the cultures, and 5 of these eventually showed cytopathic effects leading to cell death. Three of the strains were tested for pathogenicity in pigs at various passage levels. Each showed evidence of modification in virulence after a relatively few passages in PK cells. In one case modified virus produced resistance to challenge with homologous virulent virus. All strains rendered the PK cultures capable of hemadsorption of pig erythrocytes.  相似文献   

9.
非洲猪瘟(ASF)目前是生猪产业最重要的猪病。因为非洲猪瘟病毒(ASFV)本身的复杂性,以及和宿主相互作用的复杂性导致ASF已经被报道一百年了,还没有商业化疫苗。理想的ASF疫苗不仅要有好的免疫保护性,更重要的是其安全性,同时如果能区分野毒感染和疫苗接种,能在适合的高质量GMP车间进行稳定低价的生产,能用于不同物种就更好了。ASF灭活疫苗研制这条道路似乎不通;亚单位疫苗、DNA疫苗、病毒活载体疫苗的免疫保护能力不足;主要包含自然缺失、传代致弱、基因工程缺失的减毒活疫苗在免疫保护方面体现了非常大的优势,但是其潜在的持续感染风险,会造成猪只的副作用,包括皮肤溃烂、关节炎,导致神经系统、呼吸系统的损伤等问题非常值得警惕。复制缺陷单周期病毒能有效地解决减毒活疫苗的安全性问题,似乎是一个值得尝试的ASF疫苗研制方向,尽管存在难以确定缺失复制相关的基因,以及难以构建能高效表达缺失基因编码蛋白,且能让单周期病毒稳定大量生产的辅助细胞系。我国针对ASFV强毒的精准剔除策略,未注册非法弱毒苗造成临床严重损失,以及一些不使用疫苗但成功从国家层面净化ASF的案例,让我们认识到针对ASF基础研究的重要性。同时至少目前阶段,ASF的防控不一定要借助疫苗,更多的要做好生物安全管控和区域ASF净化。尽管ASF疫苗研制困难重重,但针对ASF理想型疫苗的研制也应该持续进行下去,未来可能作为ASF防控的一个突破点。  相似文献   

10.
Direct diagnosis of swine influenza infection by an indirect immunofluorescence technique using anti-nucleoproteine monoclonal antibody was compared with virus isolation. Five 8-week-old pigs were inoculated with 2 x 10(7) EID50 of strain A H1N1Sw/4115/85. Clinical signs developed in only three pigs. Antigen was detected in nasal epithelial cells obtained from all animals the first day after inoculation; the antigen was detected in one pig 6 days after the infection. Fluorescence was present in the nucleus, nucleolus and cytoplasm of infected cells. The indirect immunofluorescence test was specific and as sensitive as virus isolation in embryonated eggs, allowing a rapid diagnosis that could be achieved within hours.  相似文献   

11.
非洲猪瘟病毒无标签p30-ELISA抗体检测方法的建立及应用   总被引:1,自引:1,他引:0  
非洲猪瘟(African swine fever, ASF)是由非洲猪瘟病毒(African swine fever virus, ASFV)引起猪的一种急性、热性、出血性、高度接触性传染病,临床症状以败血症、皮炎和关节炎为特征,高发病率和高死亡率。为建立临床检测ASFV抗体的间接ELISA检测方法,本研究扩增了ASFV-CP204L基因,通过pET-30a原核表达系统表达p30蛋白,使用Ni-NTA纯化表达产物,通过肠激酶切除外源性蛋白,得到无His-组氨酸标签的p30蛋白,以此为诊断抗原,建立间接ELISA方法。结果显示:表达的无标签p30重组蛋白大小约为30 ku,与ASF阳性猪血清具有较好的反应原性;确定ELISA抗原包被浓度为1 μg·mL-1,根据ROC曲线下面积确定S/P值>0.398判定为阳性,批内、批间变异系数均<10%;与PCV2、CSFV、PRV-gE、PRRSV阳性血清无交叉反应与INGENASA商品化试剂盒总符合率为97.78%。用该方法分别检测标准阳性血清、动物感染试验血清和收集的区域性临床血清644份,该方法最低可检测到1∶512倍稀释的标准阳性血清样品;检测感染动物血清,其中80%(4/5)的试验动物在第10天抗体为阳性。644份临床猪血清样品中抗体阳性率为7.61%,其中,母猪、后备母猪、仔猪、保育猪和育肥猪抗体阳性率分别为3.03%、0%、4.94%、7.55%和28.7%。本试验建立的ASFV-p30间接ELISA方法具有良好的特异性、灵敏度和重复性,可应用于ASFV的抗体检测,为ASF的诊断和流行病学调查提供了技术手段。  相似文献   

12.
The K strain of Aujeszky's disease virus (ADV) grown in Vero cells was used to vaccinate pigs. Following intramuscular inoculation, the pigs remained healthy, no vaccine virus was excreted and virus could be detected only at the inoculation site. One inoculation gave good protection against challenge with a virulent strain of ADV, and the amount of virulent ADV excreted was geatly curtailed. Following vaccination only low leads of serum neutralizing antibody were detected (geometric mean titre 1/2), but three weeks after challenge very high levels were found (GMT 1/1773). Intranasal vaccination gave similar results. There was minimal excretion of vaccine virus. The clinical reaction on challenge was less severe than in the intramuscularly challenged group, although lower antibody levels were detected three wekks following challenge (GMT 1/483). A field trial, using this strain given subcutaneously, indicated that one inoculation of this vaccine is effective.  相似文献   

13.
In this study, an intranasal immunization strategy was set up in maternally immune pigs in order to protect them not only clinically but also virologically. Two genetically engineered Aujeszky’s disease virus (ADV) strains, Kaplan gE?gI? and Kaplan gE?gC?, were used for intranasal immunization. Both strains were safe for 4-week-old pigs. A single intranasal inoculation of 106.0 TCID50 of Kaplan gE?gI? and Kaplan gE?gC? at 4 weeks of age in the presence of moderate titres of maternally derived antibodies (SN titres: 12–16) reduced the amount of weight loss, fever and virus excretion upon challenge 6 weeks later. In a second experiment, the effect of an additional intramuscular booster with three different commercial vaccines (containing attenuated Bartha or NIA3-783 or inactivated Phylaxia; all suspended in an oil-in-water emulsion) at 10 weeks of age was evaluated. One month after the last intramuscular booster, between five and seven pigs from each group were selected for challenge. All intranasally/intramuscularly immunized pigs showed a significantly better clinical and virological protection after challenge than the single intranasally immunized pigs. In the double immunized group, the protection was better when Kaplan gE?gC? was used for the intranasal priming (only two of 14 pigs excreted virus with a duration of 4 days) than when Kaplan gE?gI? was used (13 of 18 pigs excreted virus with a duration ranging from 1 to 4 days). The virological protection was not influenced by the type of vaccine used for booster vaccination. Because the intranasal/intramuscular immunization approach is very compatible with current pig movements on farms and pigs with moderate levels of maternally derived antibodies can effectively be immunized, it can be considered as a good alternative to intramuscular/intramuscular vaccinations especially in regions with a high ADV prevalence.  相似文献   

14.
The primary objective of the study was to determine strain specificity of the immune response of pigs following vaccination with selected strains of porcine reproductive and respiratory syndrome virus (PRRSV). The experimental design included five groups (I through V, six pigs per group) free of antibody for PRRSV at the beginning of the experiment (day 0). On day 0, groups III, IV, and V were vaccinated with attenuated versions of PRRSV strains 8, 9, and 14, respectively. On day 21, the immunity of group II (non-vaccinated/challenged controls) and groups III, IV, and V was challenged by exposing each pig to a composite of the virulent versions of these same three strains. On day 35, all pigs, including non-vaccinated/non-challenged pigs of group I, were necropsied. Lungs and selected lymph nodes were examined for lesions. Serum samples obtained at weekly intervals throughout the study and lung lavage fluids obtained at necropsy were tested for the presence of PRRSV and its strain identity. Before challenge the strain of PRRSV identified in the sera of vaccinated pigs was always that with which the particular pig had been vaccinated (i.e. homologous strain), whereas, with one exception, only heterologous strains were identified after challenge. This apparent strain exclusion as a result of vaccination was statistically significant (P = 0.004). The tendency for heterologous strains to predominate after challenge suggests that a pig's immune response to PRRSV has some degree of strain specificity. Whether this finding has any clinical relevance in regard to immunoprophylaxis remains to be determined.  相似文献   

15.
In this study, an intranasal immunization strategy was set up in maternally immune pigs in order to protect them not only clinically but also virologically. Two genetically engineered Aujeszky's disease virus (ADV) strains, Kaplan gE-gI- and Kaplan gE-gC-, were used for intranasal immunization. Both strains were safe for 4-week-old pigs. A single intranasal inoculation of 10(6.0) TCID50 of Kaplan gE-gI- and Kaplan gE-gC- at 4 weeks of age in the presence of moderate titres of maternally derived antibodies (SN titres: 12-16) reduced the amount of weight loss, fever and virus excretion upon challenge 6 weeks later. In a second experiment, the effect of an additional intramuscular booster with three different commercial vaccines (containing attenuated Bartha or NIA3-783 or inactivated Phylaxia; all suspended in an oil-in-water emulsion) at 10 weeks of age was evaluated. One month after the last intramuscular booster, between five and seven pigs from each group were selected for challenge. All intranasally/intramuscularly immunized pigs showed a significantly better clinical and virological protection after challenge than the single intranasally immunized pigs. In the double immunized group, the protection was better when Kaplan gE-gC- was used for the intranasal priming (only two of 14 pigs excreted virus with a duration of 4 days) than when Kaplan gE-gI- was used (13 of 18 pigs excreted virus with a duration ranging from 1 to 4 days). The virological protection was not influenced by the type of vaccine used for booster vaccination. Because the intranasal/intramuscular immunization approach is very compatible with current pig movements on farms and pigs with moderate levels of maternally derived antibodies can effectively be immunized, it can be considered as a good alternative to intramuscular/intramuscular vaccinations especially in regions with a high ADV prevalence.  相似文献   

16.
Summary

The pathogenicity and pathogenesis of Lelystad virus was studied in six 6‐day‐old SPF piglets. A third passage of the agent was propagated on porcine alveolar macrophages and intranasally inoculated into pigs. Pigs were killed at hours 24, 48, 60, and 72, and on days 6 and 8 after inoculation. From day 2 on pigs developed diffuse interstitial pneumonia with focal areas of catarrhal pneumonia, and from this day on splenic red pulp macrophages were enlarged and vacuolated. Lelystad virus was re‐isolated from the lungs of infected pigs from day 2 after inoculation. Lelystad virus antigens were detected by immunohistochemical techniques in bronchiolar epithelium and alveolar cells, and in spleen cells of infected pigs from day 2 after inoculation. Ultrastructural examination of tissues by electron microscopy revealed degenerating alveolar macrophages and epithelial cells in lungs and nasal mucosa, with excessive vacuolation of the endoplasmic reticulum.

Although the respiratory tract seems to be the target organ for this virus, macrophages in other organs, such as the spleen, can also be infected. This preference for macrophages may impair immunological defences.  相似文献   

17.
Antibody levels in post-infection sera from a pig inoculated with a low virulent strain of classical swine fever virus (Hannover 62) and in sera from two pigs inoculated with another low virulent strain (Spielbach 66) and from an in-contact pig were assayed by complement fixation and immunofluorescence using classical swine fever virus (ALD strain) and bovine virus diarrhoea virus (UG 59 strain) as antigens. The complement fixation test used was modified by addition of a preparation of porcine Glq to the complement and by mercaptoethanol treatment of the immune serum before use. The mercaptoethanol treatment of the immune serum resulted in complete elimination of a haemolytic prozone often seen with porcine immune sera.In the sera from the inoculated animals complement-fixing antibodies appeared earlier than neutralizing antibodies. A few weeks after inoculation there was a correlation between the presence of complement-fixing and neutralizing antibodies.During the entire observation period of 13 weeks it was not possible to demonstrate complement-fixing or neutralizing antibodies in serum from a pig exposed to infection by contact with the two pigs inoculated with the Spièlbach 66 strain of classical swine fever virus.  相似文献   

18.
The production of interferon by pigs in response to viral and synthetic inducers was studied. The inducers used included polyriboinosinic-polyribocytidylic acid (Poly I:C), swine influenza virus and pseudorabies virus. Following intravenous inoculation of pigs with the inducers, sera were examined for interferon by the plaque-reduction method in porcine kidney (PK15) cell cultures using vesicular stomatitis virus as the challenge inoculum. It was shown that pigs can produce interferon in response to each of these inducers. The pseudorabies virus used in this investigation was found to be a better interferon inducer than the swine influenza virus.

The interferon produced in pigs was identified as an interferon because it was pH stable, non-dialyzable, sensitive to trypsin, non-sedimentable and possessed broad-spectrum antiviral activity as well as host-species specificity.

  相似文献   

19.
Chronic pneumonia developed in 14 pigs inoculated with an attenuated strain of African swine fever (ASF) virus. The pathogenesis of the pneumonia was as follows: (1) Interalveolar septums became thickened by accumulation of lymphocytes and monocytes; (2) lung developed focal areas of lymphocytes and macrophages; (3) necrosis began abruptly in these foci, beginning with the cells in the alveolar lumens, developing in centrifugal direction, and eventually affecting all structures in its path; (4) necrotic tissue became calcified; and (5) a mantle of mononuclear cells (including plasma cells) and fibrous tissue formed around the necrotic area. Viremia occurred in the 14 pigs at postinoculation day (PID) 14, and precipitating antibody was increased significantly at PID 58.  相似文献   

20.
用固定细胞阻断酶联免疫吸附试验(ELISA)对来自丹麦猪的180份血清进行了猪传染性胃肠炎病毒(TGEV)和猪呼吸道冠状病毒(PRCV)感染的鉴别诊断,共检出了PRCV抗体阳性血清107份(59.4%),TGEV抗体阳性血清0份。同时也检测了一些来自国内不同TGEV感染类型的猪场血清。该鉴别诊断方法在我国的建立和应用为从PRCV阳性国家进口猪的TGE的检疫提供了一条有效途径。  相似文献   

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