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禽巴氏杆菌荚膜多糖-蛋白载体抗原的研究 总被引：4，自引：1，他引：3
用EDC法和CNBr法分别交联禽巴氏杆菌荚膜多糖(CPS)与破伤风类毒素(TT),经Sephadex-G150柱层析和薄层层析鉴定表明;两种方法均获得一种大分子的CPS-TT结合物,该结合物免疫鸡的血清经二巯基乙醇(2-ME)处理后用间接血凝试验(IHA)检测出较高滴度的IgG抗体和一定量的IgM抗体,抗体消长情况监测结果;IgG抗体的峰值(4.88)在免疫后第21天,其后缓慢下降,持续至第24周左右。IgM抗体的峰值(4.26)在第14天,其后陡降,至第8周降至阴性血清水平,与对照组差异极显著(P<0.01)。第8周和第24周时测出较强的二次反应和回忆反应。重复试验测出的IgG、IgM抗体滴度与本试验结果相近,第4周时攻毒保护率分别为100％和75％,且IgG抗体滴度与攻毒保护率的相关性较好,1:16以上可获得全保护。试验结果表明交联方法稳定,重复性较好,制备的CPS-TT载体抗原实现了CPS由Ti抗原向Td抗原的转换。为研究一种新型的禽霍乱载体菌苗提供了理论依据和实验手段。 相似文献
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不同培养工艺制备的禽多杀性巴氏杆菌菌苗抗原的免疫原性比较试验 总被引：1，自引：0，他引：1
为比较不同培养工艺制备的禽多杀性巴氏杆菌抗原的免疫原性,对禽多杀性巴氏杆菌标准强毒株C48-1株采用固体表面培养法和液体高密度发酵法制备疫苗并进行了免疫对比试验.结果表明,固体表面培养法制备的抗原菌体形态结构较均一,抗原成分稳定一致,并且含有较丰富的荚膜;而液体高密度发酵法制备的抗原形态结构大小不一,抗原成分不够稳定一致,含荚膜也较少.将两种方法制备的抗原按相同工艺制备成禽霍乱蜂胶灭活疫苗,效力检验结果显示二者差异极显著,固体表面培养法制备的疫苗免疫后14 d和90 d时保护率均为80%～100%,平均93%;液体高密度发酵法制备的疫苗免疫后14 d和90 d时保护率分别为40%～60%、60%～80%,平均为53%和67%. 相似文献
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禽多杀性巴氏杆菌是引起鸡、鸭和鹅等多种禽类的一种急性败血性传染病病原菌。制成的菌苗在安全性或免疫性等方面还不十分理想。近年来国外不少学者曾就禽巴氏杆菌细胞壁荚膜及免疫性进行了研究,我国对该菌菌体及其荚膜抗原性本身的研究还不多。我们曾用2.5％NaCl溶液自禽多杀性巴氏杆菌C_(48-1)强毒株浸提出一种保护性荚膜抗原,已经证明这种粗提荚膜抗原对鸡、鸭安全和具有良好的免疫保护力。有关对该菌的超微结构及其提取菌体胞壁的荚膜后的形态变化,目前在国内还未见有正式报道。为了给今后免疫研究工作提供依据,我们曾将我国的一株禽多杀性巴氏杆菌C_(48-1)强毒株用 相似文献
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hyaD为A型多杀性巴氏杆菌荚膜多糖合成相关基因,为探讨该基因对多杀性巴氏杆菌毒力及其免疫保护特性的影响,本研究利用同源重组方法,构建了牛源A型多杀性巴氏杆菌CQ2株（PmCQ2）的hyaD基因缺失株（ΔhyaD）。结果发现,与野生株相比,ΔhyaD的荚膜产生量及其感染后在脏器中的细菌定殖量均显著下降,其毒力显著降低。细胞试验发现,ΔhyaD更易黏附于巨噬细胞,被吞噬数量显著多于野生株,致使巨噬细胞相关炎性因子表达显著上调。hyaD基因的缺失,可调控与荚膜合成、LPS合成转运、铁转运等相关的基因表达显著下调,促使相关保护性抗原基因表达显著上调。以制备的PmCQ2株和ΔhyaD株灭活苗免疫小鼠（加强免疫1次）,免疫后第21天分别采用同源和异源多杀性巴氏杆菌攻毒,ΔhyaD株免疫小鼠肺组织感染后24 h无明显或轻微病理损伤,对牛源A型、B型和F型多杀性巴氏杆菌的免疫保护率分别为100%、100%和80%,对兔源、猪源和禽源A型多杀性巴氏杆菌的免疫保护率分别为90%、100%、100%;而野生株PmCQ2除对牛源A型多杀性巴氏杆菌的保护率在80%以上外,对牛源B型和F型及兔、猪、禽源A型多杀性巴氏杆菌均无明显交叉保护作用。研究结果表明,hyaD基因可通过调控荚膜产生及毒力相关因子表达影响菌株毒力;hyaD基因缺失可调控相关交叉保护性抗原表达,赋予菌株交叉免疫保护特性。该研究为多杀性巴氏杆菌通用型疫苗的研发提供了参考。 相似文献
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抗牛多杀性巴氏杆菌高免卵黄抗体IgY的制备及间接血凝检测方法的建立 总被引：1，自引：0，他引：1
本研究通过自制牛源荚膜血清A型多杀性巴氏杆菌灭活菌苗免疫产蛋鸡,采用卡拉胶结合硫酸铵沉淀法提取卵黄抗体IgY,并采用间接血凝方法检测抗体效价。结果表明,抗原致敏红细胞的最佳浓度是800μg/mL,免疫后第7周抗体效价达到高峰,效价为1：1024,高效价持续5周开始下降,测定提取的IgY浓度为8.258mg/mL,无菌检测及动物安全性实验表明制备的卵黄抗体安全可靠。本研究制备了抗牛多杀性巴氏杆菌卵黄抗体,为防治由荚膜血清A型多杀性巴氏杆菌所致的犊牛肺炎提供了新的手段。 相似文献
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以家畜伊氏锥虫弱毒株免疫家兔12只,然后进行攻虫试验。其中11只被保护,1只发病死亡。以BA-ELISA检测家兔免疫后及攻虫后特异性IgG、IgM的变化,发现免疫后IgG、IgM均于第21d达到峰值;攻虫后IgM于第6d达到峰值,IgG于第9d达到峰值。攻虫后,随着免疫组家兔体内特异性IgG、IgM的不断上升,其血液内的锥虫也同时被清除。体外中和试验也表明,特异性抗体对锥虫的感染力具有明显的抑制作用。 相似文献
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《中国兽医学报》2015,(4):549-552
分别以活菌浓度为109(A组)、106(B组)ccu/mL的(MG)F株及生理盐水(C组)点眼接种SPF鸡,采用流式细胞技术及ELISA方法对免疫前后外周血淋巴细胞(PBL)中Bu-1a+B、TCRγδ+CD3+T细胞比例及血清IgM、IgG、IgA的动态变化规律进行研究。结果显示各组Bu-1a+B/PBL于免疫后5、7d由高至低依次为A、B和C组,且各组差异显著(P0.05);A、B组TCRγδ+CD3+T/PBL于免疫后3d略有升高且比值较接近,都高于C组,但差异不显著(P0.05);各组IgM于免疫后7、14和21d差异显著(P0.05),IgG于免疫后14、21和28d差异显著(P0.05),IgA于免疫后28d差异显著(P0.05),由高至低依次为A、B、C组。研究结果表明,免疫(MG)F株可较好的提高Bu-1a+B、血清IgM、IgG、IgA介导的体液免疫作用,且(MG)F株活菌浓度与Bu-1a+B/PBL比值、血清IgM、IgG、IgA浓度呈正相关;但免疫(MG)F株不能显著提高鸡外周血TCRγδ+CD3+T/PBL比值,且(MG)F株活菌浓度与TCRγδ+CD3+T/PBL比值无明显相关性。 相似文献
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Takashima H Sakai H Yanai T Masegi T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2001,63(2):171-174
To detect serum antibody against Pasteurella multocida (P. multocida) in infected rabbits. a modified immunoperoxidase assay was applied. An outbreak of P. multocida infection in rabbits started from sudden death. The infected rabbits had severe fibrinous and purulent pneumonia with hemorrhage, and a large number of P. multocida (A:12) was isolated from the trachea and lungs of the animals. Antibodies of IgM and IgG to P. multocida were assessed by immunohistochemical staining using the sera of the animals as primary antibodies and applying them to formalin-fixed, paraffin-embedded sections of P. multocida attached to calf fibrin. IgM antibodies to P. multocida were first detected 7 days after the onset of the disease. IgG antibodies began to rise on the 7th or 14th day. These results suggested that the modified immunoperoxidase assay could detect antibodies against P. multocida. 相似文献
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Prado ME Prado TM Payton M Confer AW 《Veterinary immunology and immunopathology》2006,111(3-4):301-307
The dynamics and duration of maternally derived antibodies as well as the onset of acquired immunity against Mannheimia haemolytica and Pasteurella multocida in range-pastured beef calves were investigated. Two groups of unvaccinated cattle were used in this study. Serum antibody responses were measured by enzyme-linked immunoassay for antibodies of the IgG1, IgG2 and IgM isotypes binding M. haemolytica whole cells (WC) or leukotoxin (LKT) and P. multocida outer membrane proteins (OMPs). Comparisons of mean antibody responses to M. haemolytica LKT and WC and P. multocida OMPs were made within each group. Maternally derived antibodies against M. haemolytica and P. multocida reached lowest levels at 30-90 days after birth. Calves began production of antibodies against M. haemolytica and P. multocida between 60 and 90 days of age in both groups. Based on the results of this study, in beef herds vaccinated against M. haemolytica and/or P. multocida, it may be best to vaccinate calves around 3 months of age. In contrast, beef calves from unvaccinated herds might benefit from vaccination at 4 months of age. 相似文献
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Vaccination of neonatal colostrum-deprived calves against Pasteurella haemolytica A1. 总被引：1，自引：1，他引：0 
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下载免费PDF全文 Colostrum-deprived Holstein calves were vaccinated at 2 and 4 wk of age with a Pasteurella haemolytica A1 culture supernatant vaccine to determine whether active immune responses and protection could be induced in this age group in the absence of maternal antibodies. All calves responded to vaccination with high titers of IgM antibodies to capsular polysaccharide within 1 wk of primary vaccination. Mean titers of IgG1 and IgG2 antibodies to this antigen increased significantly by 2 wk after secondary vaccination, but peak antibody titers were low. All of the vaccinated calves seroconverted with production of leukotoxin-neutralizing antibodies, but peak antibody titers were low. Vaccinated calves experienced considerable lung damage after experimental challenge, but survival rate, clinical scores, and percent lung involvement were significantly better than those of control (placebo-injected) calves. 相似文献
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R A Oellermann P Carter M J Marx 《The Onderstepoort journal of veterinary research》1976,43(4):185-188
The IgM and IgG response of mice to sheep erythrocytes (SRBC) and bluetongue virus (BTV) was determined by means of haemolytic plaque assays. Maximum primary IgM response to SRBC occurred after 4 days but declined rapidly to 4% of the maximum by Day 9. A lag period of about 2 days was observed in the appearance of IgG haemolytic plaque-forming cells (PFC) but they reached a maximum after 6-9 days. Secondary immunization resulted in the stimulation particularly of IgG PFC and from Day 6 onwards IgG predominated in the immunological response. The IgM response to BTV was remarkably similar to that observed when SRBC were used as antigen. IgG PFC, however, appeared within a day of the IgM, reaching a peak on Days 4-5. From then onwards, IgG PFC predominated in the response. At BTV concentrations of up to 10 mug per mouse, the virulent strain of BTV type 3 produced the weakest response. At higher antigen concentrations there was very little difference in the response to the serotypes tested, although the virulent strain of BTV type 4 tended to produce the strongest response. 相似文献
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OBJECTIVE: To determine the seroprevalence of antibodies to gram-negative core antigens (GNCA) in specific-pathogen-free (SPF) rabbits (ie, free of Pasteurella multocida) and rabbits of undefined bacterial status (conventional). SAMPLE POPULATION: Serum samples were obtained from 7 groups of rabbits. The SPF rabbits comprised 2 adult groups and 1 immature group, whereas the 4 groups of conventional rabbits were all adults. PROCEDURE: A seroprevalence survey was conducted on rabbit sera for antibodies against GNCA, using an Escherichia coli J5 antigen-capture ELISA. RESULTS: Collective geometric mean titer (GMT) of adult rabbits was 1:6,463. The GMT of each of the 6 groups of adult rabbits was 1:956, 1:1,133, 1:4,525, 1:5,338, 1:7,669, and 1:25,600. Titers of populations differed significantly. CONCLUSION: Data analysis revealed there were anti-GNCA antibodies in rabbits. Similar to other species, the prevalence of IgM and IgG anti-GNCA antibodies increased with age. The IgG response was more marked than the IgM response. The SPF rabbits had lower IgG anti-GNCA titers than conventional rabbits, indicating possible cross-reactive epitopes between P multocida and Enterobacteriaceae. Rabbits with the highest anti-GNCA titers were those used in polyclonal antibody production, possibly stemming from endotoxin contamination of antigen or adjuvant. CLINICAL RELEVANCE: The possible cross-reactive antibodies directed at homologous wall components of Pasteurellaceae and Enterobacteriaceae could prove to be a possible heterotypic vaccination strategy for the protection of rabbits against pasteurellosis. Investigators should determine whether antigen impurity (endotoxin contamination) influences epitope focus during polyclonal antibody production and whether it affects sera variability among rabbits. 相似文献
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An indirect enzyme-linked immunosorbent assay (ELISA) was developed to measure humoral antibody responses of chickens against Pasteurella multocida. A standard indirect hemagglutination (IHA) test was used to compare serologic results with those of ELISA. The ELISA was also used following challenge with P. multocida to compare the efficacy of three commercial fowl cholera vaccination regimens. Although antibody titers measured by ELISA and IHA were highly correlated, ELISA was at least twice as sensitive as IHA. Antibody measured by ELISA and IHA also correlated significantly with protection against P. multocida challenge. No mortality occurred in any of the three vaccinated challenged groups. However, control unvaccinated chickens experimentally infected with P. multocida developed signs of acute pasteurellosis and died by the 10th day post-challenge. Impression smears made of hepatic tissue from all chickens were stained (Wright's stain), and typical bipolar rods characteristic of Pasteurella were identified in smears from unvaccinated challenged controls only. 相似文献
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C N Gresham A W Confer L J Bush J A Rummage 《American journal of veterinary research》1984,45(11):2227-2230
A survey of antibody to Pasteurella haemolytica and P multocida, using a fluorometric immunoassay, was conducted on sera collected from 264 dairy cattle from 3 herds. Serum antibody titers to P haemolytica were 0 to 270 with low titers (less than 25) seen in 48.1% of the cows and heifers. Serum antibody titers to P multocida were 0 to 380 and the frequency of distribution of these titers were more even than for P haemolytica. Mean serum antibody titers to P haemolytica were significantly (P less than 0.005) higher in cattle from an open dairy herd when compared with those from 2 closed herds. Antibody titers to these organisms was determined in 7 colostrum samples. Pasteurella haemolytica antibody titers varied, depending on the whey separation technique used. Passive transfer of colostrum-derived antibody in 5 neonatal calves resulted in a maximum mean serum antibody titer at 20 hours after birth for P haemolytica and at 8 hours after birth for P multocida. Serum titers were higher overall for P multocida than for P haemolytica. Serum titers for P haemolytica declined rapidly. A significant (P less than 0.05) increase in antibody to P multocida was observed at 5 days of age. 相似文献
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Rabbit pasteurellosis: induced disease and vaccination 总被引：1，自引：0，他引：1
Pasteurellosis was induced in rabbits by conjunctival inoculation with 2 strains of Pasteurella multocida. The LD50 of strain P1062 (a bovine isolate) was 10(5.1) colony-forming units and that of strain P1059 (a turkey isolate) was 10(5.5) colony-forming units. Pasteurella-free rabbits were vaccinated IV or mucosally with boiled cells of P multocida or a cross-reactive uridine diphosphogalactose epimerase-deficient mutant of Escherichia coli J5. In rabbits challenge exposed with P multocida strain 1062 or 1059, homologous P multocida strain gave the best protection against fatal bacteremia. Partial protection was provided by J5; mucosal routes of vaccination (aerosol or conjunctival) gave better protection than did the IV route. Serum antibody titers were lower in rabbits vaccinated by mucosal routes than in those vaccinated IV. Cross-reactive IgG and IgM titers to P multocida were demonstrated when rabbits were vaccinated with J5. On the basis of bacteriologic examination of nasal secretions, rabbits that died were considered culture positive sooner than were those that survived. On the basis of bacteriologic examination of blood, rabbits that died were considered culture positive, and those that survived were considered culture negative. Seemingly, heat-stable antigens were protective, the cross-reactive E coli J5 mutant (with only core lipopolysaccharide) provided partial protection against pasteurellosis, and the mucosal route was somewhat useful for cross-protective immunization. 相似文献
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禽霍乱免疫母鸡血清抗体和蛋黄抗体消长规律及其相互关系的研究 总被引：3，自引：0，他引：3
通过对禽霍乱菌苗免疫产蛋母鸡的血清和蛋黄的抗体定时检测后发现，免疫母鸡的血清抗体和蛋黄抗体的升降趋势基本一致（ｒ＝０．９４），但后者较前者迟后３～６ｄ；血清抗体和蛋黄抗体的滴度分别在加强免疫后的１～３ｄ和３～６ｄ都有不同程度下降，加强免疫前的滴度越高，免疫后的滴度下降幅度越大；蛋黄抗体水平普遍比血清抗体水平低，两者间的差异极显著（Ｐ＜０．０１），这可能是与禽多杀性巴氏杆菌本身的主要抗原成分（ＴＩ抗原）在鸡体内主要产金ＩｇＭ有关。 相似文献