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1.
应用间接ELISA方法,研究经嗜水气单胞菌(Aeromonas hydrophila)疫苗浸泡免疫后鳜(Siniperca chuatsi)皮肤黏液中抗体消长规律,以揭示鳜局部黏膜免疫在免疫保护中的作用及浸泡免疫的保护效果。结果表明,鳜皮肤黏液中抗体滴度在免疫后第7天达到峰值211,抗体从开始形成到消失持续21 d;添加佐剂(IMS1312、葡聚糖、莨菪碱、食盐)可以提高鳜皮肤黏液及血清中抗体滴度和相对免疫保护率,其中添加IMS1312组免疫保护率最高,达77.8%,免疫保护与抗体滴度成正相关(R2=0.79,P<0.05);以溶菌酶为代表的非特异性免疫在免疫后1周内即抗体未形成时,对鱼体起主要保护作用。此外,通过比较鳜血清与皮肤黏液中的抗体消长规律,发现血清中抗体滴度峰值出现时间较迟(第14天),抗体持续时间较长(42 d),初步推断鳜皮肤黏膜中可能存在相对独立于全身免疫系统的局部黏膜免疫系统。 相似文献
2.
蜂胶佐剂对异育银鲫嗜水气单胞菌灭活疫苗的免疫增强试验 总被引：2，自引：0，他引：2
本文就蜂胶佐剂对鱼类细菌性疫苗的免疫增强作用进行了首次评价。试验以异育银鲫嗜水气单胞菌灭活疫苗为对象,比较了蜂胶佐剂、铝胶佐剂和无佐剂三种疫苗对银鲫的免疫效果。经抗体监测表明,免疫接种后第2周,蜂胶佐剂组银鲫已检测到血清抗体,随后抗体效价迅速上升,第4周达到峰值2^5：铝胶佐剂组和无佐剂组第3周检测到抗体．铝胶佐剂组第5周达到峰值2^5,无佐剂组第4周达到峰值2^4。银鲫免疫后第5周进行攻毒保护试验,发现蜂胶佐剂组、铝胶佐剂组和无佐剂组的免疫保护率分别为77．3％．65．0％和56．3％。试验结果表明,蜂胶佐剂能有效增强异育银鲫细菌性灭活疫苗的免疫保护力。 相似文献
3.
山莨菪碱提高嗜水气单胞菌灭活疫苗浸泡免疫鲫的效果 总被引：4，自引：3，他引：1
为研究山莨菪碱作为佐剂在细菌灭活疫苗浸泡免疫中的作用,将山莨菪碱和嗜水气单胞菌全菌灭活疫苗联合浸泡免疫异育银鲫,首次浸泡免疫7d后加强免疫1次.通过实时荧光定量PCR检测第2、4、7、11、14和21天脾脏中IgM、IL-1β、C3、C-凝集素以及溶菌酶的mRNA表达量,并在第21天进行同源菌株活菌攻击实验.结果显示,山莨菪碱组第4天IgM和IL-1 β的表达量达到最高值,分别为28.3和332.7;而无佐剂疫苗组第11天IgM和IL-1β达到最高值,分别为56.1和791.8.山莨菪碱组的补体C3、C-凝集素以及溶菌酶的表达量都显著高于无佐剂组和对照组,且表达持续时间长.活菌攻击实验表明山莨菪碱组的相对免疫保护率可达80.0％,显著高于无佐剂组的56.0％.结果表明,山莨菪碱与嗜水气单胞菌灭活疫苗共同浸泡免疫银鲫,可以增强脾脏中IgM、IL-1β、C3、C-凝集素以及溶菌酶基因表达,提高银鲫相对免疫保护率. 相似文献
4.
为了探讨迟缓爱德华菌菌蜕疫苗预防鳗鲡爱德华菌病的可行性,实验采用腹腔注射、口服、浸泡3种途径探讨了迟缓爱德华菌菌蜕疫苗对欧洲鳗的免疫保护效果。结果表明,免疫后欧洲鳗血清抗体效价均明显升高,但菌蜕疫苗(ETG)组与福尔马林灭活疫苗(FKC)组间血清抗体效价无显著性差异。欧洲鳗注射、口服、浸泡ETG组的相对免疫保护率分别为75.0%、52.5%、37.5%,分别高于FKC组的55.0%、40.0%、32.5%,且ETG组欧洲鳗死亡时间明显推后,表明菌蜕疫苗的免疫保护效果优于福尔马林灭活疫苗。易操作、对鱼体应激少的ETG疫苗口服免疫欧洲鳗获得了52.5%的免疫保护率,在预防鳗鲡爱德华菌病中具有良好的应用前景。 相似文献
5.
嗜水气单胞菌疫苗的初步研究 总被引：1，自引：1，他引：1
用来源于乌鳢腹水病的嗜水气单胞菌，制成菌体抗原与可溶性抗原，分6组注射接种湘鲫，检测供试鱼血清中凝集抗体效价。结果表明：菌体抗原组与可溶性抗原组分别在第2l天和第14天达到峰值，然后随免疫时间逐渐下降。加弗氏不完全佐剂的菌体抗原组与可溶性抗原组分别在第2l天与第35天分别达到峰值。并保持此水平。对鱼体免疫35天后的保护率进行测定，结果表明：疫苗组为100％，佐剂组为30％．生理盐水对照组为0％。 相似文献
6.
白鲢对嗜水气单胞菌疫苗的免疫应答 总被引：4，自引：0，他引：4
用嗜水气单胞菌9120株制备了鱼类出血性败血症疫苗。用该疫苗分二组对白鲢进行免疫，检测免疫后白鲢血清中的抗体水平，其中加佐剂免疫组获得了较高滴度的凝集抗体，不加佐剂免疫组产生的凝集抗体滴度较低，但用间接血球凝集试验可以检出。 相似文献
7.
为研究罗非鱼源无乳链球菌溶血素(Hemolysin,Hly)对鱼体的免疫保护作用,根据已获得的无乳链球菌ZQ0910全基因组序列设计引物扩增hly基因,定向克隆于原核表达载体p ET-28a中,构建原核重组质粒p ET-28a-hly,经IPTG诱导表达后,制成亚单位疫苗免疫吉富罗非鱼,并分析疫苗的免疫保护力。结果显示,hly基因产物大小1335 bp,编码444个氨基酸,经测序与Gen Bank报道的链球菌属Hly氨基酸序列同源性可达99%。经IPTG诱导表达后,SDS-PAGE分析可见一条51.7 k D的特异条带;Western blotting分析结果说明表达的Hly蛋白能与His-Tag单抗特异性结合;制备的亚单位疫苗免疫鱼体后第14天即可检测到抗体产生,并在第28天达到峰值,抗体效价为1∶4096,免疫保护率为70%。由此证实,该亚单位疫苗有望成为预防由无乳链球菌引起的罗非鱼链球菌病的基因工程类疫苗。 相似文献
8.
嗜水气单胞菌灭活疫苗浸泡后鳜IgM基因表达量和抗体效价的变化 总被引：3，自引：2，他引：1
应用荧光定量PCR技术,研究了嗜水气单胞菌疫苗浸泡免疫后,鳜(Siniperca chuatsi)鳃、皮肤、脾脏和头肾中IgM基因表达量变化,同时应用ELISA检测皮肤黏液和血清中抗体滴度变化。结果显示:最早检测到IgMmRNA转录水平上调的是皮肤和鳃(第4天),而脾脏和头肾在第7天才达到高峰。IgM基因在头肾和脾脏中表达量较高(高峰值分别达到16.3和23.8),皮肤和鳃中的表达量较小(高峰值分别为4.3和8.6)。抗体效价方面,皮肤黏液中抗体滴度峰值出现时间较早(第7天),但抗体从开始形成到消失持续时间较短(28 d);血清中抗体滴度峰值出现时间较迟(第21天),但持续较长(42 d)。结果表明:浸泡免疫能够引发鳜机体和局部均出现免疫应答。从抗体滴度和IgM基因表达量方面考虑,系统免疫组织均高于黏膜免疫组织,表明前者是合成IgM的主要场所。从时间上比较,对抗原刺激最早做出应答的是皮肤黏膜和鳃,其后系统免疫才表现应答作用,这表明了局部黏膜能够在抗原入侵的早期起到抵制作用。 相似文献
9.
镉对鲫特异性免疫力的影响 总被引：2，自引：0，他引：2
研究镉对鲫特异性免疫力的影响。按大肠杆菌粗蛋白浓度3n训kg免疫鲫后，立即以Ca2^ 浓度1．25、2．5和3、75mg／kg腹腔注射染镉，在免疫染镉后的不同时间用反向间接血凝抑制法检测其抗体效价。结果显示：在试验剂量和时间范围内，鲫抗体生成的时间约为免疫后10d左右；1．25mg／kg组的反向间接血凝抑制抗体效价(RIHIAT)与对照组的RIHIAT、比较，其差异不显著；在10d、14d、21d和28d时，对照组的RIHAIT、与2．50mg／kg和3．75mg／kg组的RIHIAT、比较，其差异均有显著性，表现为镉对鲫RIHIAT始终存在着抑制作用并且有剂量-效应关系和时间-效应关系。因此Ca2^ 浓度低时对鲫体液免疫应答水平的影响不大，而Ca2^ 浓度高时对鲫体液免疫应答有抑制作用。 相似文献
10.
草鱼出血病病毒vp6核酸疫苗的免疫效果评估 总被引：2，自引：0，他引：2
为了评估草鱼出血病病毒(GCRV)vp6基因核酸疫苗的免疫效果,将vp6基因克隆进pFastBacTMDual载体杆状病毒的多角体蛋白(Ph)启动子下游,同时将团头鲂(Megalobrama amblycephala)β-actin启动子控制的vp6基因克隆进杆状病毒的P10启动子下游,获核酸疫苗载体pFastBac-FA-VP6-ph-VP6。按每尾分别注射疫苗载体10、30、60μg的剂量免疫草鱼(Ctenopharyngodon idellus)(体长14～20 cm,体质量60～120 g),同设pFastBacTMDual载体(30μg/尾)阴性对照组及空白对照组(0.4 mL/尾无菌水)。于免疫后不同时间通过RT-PCR检测免疫鱼体中vp6基因的表达,并于免疫后第14、21、28、49、70天分别通过间接凝集反应检测血液中的抗体水平,并在免疫第21天感染GCRV评估免疫保护效果。结果显示,核酸疫苗免疫草鱼后,各免疫组均有抗体产生,抗体效价在免疫后第28天达到最高;攻毒后每尾注射疫苗载体10、30、60μg组的死亡率分别为0%、0%、5%,pFastBacTMDual载体对照组和空白对照组分别为30%和100%。表明构建的核酸疫苗对草鱼病毒性出血病有较好的免疫保护效果。 相似文献
11.
Abstract. Groups of channel catfish, Ictalurus punctatus (Rafinesque). were administered Edwardsiella ictaluri twice in the form of whole outer membrane proteins (OMP) or heat-inactivated whole bacteria (IWB) orally or IWB intraperitoneally. Antibody titres and lysozyme concentrations were determined in serum, mucus, gut contents and gut washings taken 7, 14, 21 and 28 days after the second antigen administration. Bacterial killing by anterior kidney neutrophils was determined 7, 14 and 21 days after the second antigen administration. Enhanced killing of tumour targets by nonspecific cytotoxic cells (NCC) was determined 7 days after the second antigen administration. Serum antibody titres of all treatment groups were significantly increased above those of the control group throughout all sampling periods. Fish receiving oral administration of OMP or IWB exhibited maximum serum antibody titres on day 21 or 28. Antibody titres were also detected in mucus, gut washings and gut contents, but did not reach the level of those in the serum. Bacterial killing was significantly increased only on day 7, and could not be correlated to antibody titre or lysozyme concentration. Bacterial killing was found to be the result of a heat-labile serum factor. NCC activity of fish vaccinated with IWB orally was significantly higher than that of fish vaccinated with OMP orally or controls. Although intraperitoneal vaccination consistently produces higher titre antisera, the results of this study support the idea that oral vaccination can induce antibody systhesis. 相似文献
12.
MURIEL COSSARINI-DUNIER 《Journal of fish diseases》1986,9(1):27-33
Abstract. Rainbow trout, Salmo gairdneri Richardson, (average weight 100g) were vaccinated intraperitoneally (i.p.) with Yersinia ruckeri bacterin in saline or in oily adjuvant. Agglutinating antibody kinetics were followed during 445 days before challenge (1.2×107 bacteria i.p.). Fourteen days after challenge 88.5% of the control fish had died while few mortalities were observed with vaccinated fish regardless of their agglutinating titre. Protection against Y. ruckeri does not seem to be due to antibodies. 相似文献
13.
B K Gudmundsdóttir H Jónsdóttir V Steinthórsdóttir B Magnadóttir & S Gudmundsdóttir 《Journal of fish diseases》1997,20(5):351-360
Atlantic salmon were vaccinated against Aeromonas salmonicida ssp. achromogenes (Asa) by injection with three vaccines developed in our laboratory and an autogenous bacterin (IcelandBiojec.OO, IBOO) produced by a commercial vaccine producer. The humoral antibody responses to bacterial antigens were monitored by ELISA and Western blotting. The fish were challenged by infection with Asa 6 and 12 weeks post-vaccination. Protection was induced in all groups of vaccinated fish. The protection achieved was time-dependent. The autogenous bacterin, IBOO, induced a protective immune response later than our experimental vaccines. All the vaccines tested induced specific antibody response that increased between 6 and 12 weeks after vaccination. The antibody response was mainly directed against the A-layer protein, but antibodies to other bacterial components were also detected. Significant correlation was obtained between the antibody titre to extracellular Asa antigens, induced by the different vaccine preparations, and survival of vaccinated fish challenged by a virulent Asa strain. Furthermore, the detection of antibodies directed against an extracellular toxic metallo-caseinase, AsaP1, in fish sera correlated with protection. 相似文献
14.
草鱼出血病病毒VP6蛋白的原核表达、纯化及免疫效果 总被引：2，自引：2，他引：0
为了探讨草鱼出血病病毒(GCRV)VP6亚单位疫苗对草鱼出血病的免疫保护作用,将vp6基因克隆进原核表达载体pET-28a(+),构建了重组质粒pET28a(+)-VP6。SDS-PAGE和Western-blotting显示,重组VP6蛋白的分子量约为43 ku,主要以包涵体形式存在,重组蛋白占菌体总蛋白的20%左右;Ni2+柱纯化后的重组蛋白,以每尾500μg肌肉注射免疫草鱼(14～20 cm,60～120 g),并在第14、21、28、49、70天通过间接凝集反应检测抗体水平,结果显示,免疫注射后第14天可检测到鱼体产生的特异性抗体,第21天达到最高峰,第70天仍可检测到抗体的存在;免疫注射第21天人工接种GCRV,免疫后的草鱼对出血病的保护力达100%。 相似文献
15.
Abstract. A bacterin for immunization against bacterial kidney disease of salmonid fishes caused by Renibacterium salmoninarum is described. Cultures were grown in Evelyn's KDM2 medium containing 10% calf serum in a fermenter under the following conditions: pH 7.2, 15°C, 800ml/min air, 200 rev/min agitation and 5–15 days of incubation. Possible substitutes for calf serum were 10% horse serum 0.15% starch and leptospira medium. The bacterins were inactivated with 0.3% formalin and no adjuvants were used. Other tests evaluated pH-lysed bacterin, 50% concentrated bacterin and 50% concentrated pH-lysed bacterin. Juvenile rainbow trout, salmo gairdneri Richardson, were vaccinated either by intraperiotoneal (i.p.) injection, 2 min immersion or 2-step hyperosmotic infiltration. Fish were held from four to six weeks at 11°C, then challenged by i.p. injection with the homologous virulent bacterium. Fish died from days 19 to 40 after challenge. The best preparation was pH-lysed bacterin given by a single i.p. injection; hyperosmotic and immersion vaccination were not effective. Typically when 80% or more of unvaccinated controls were infected as detected by Gram stain, 10% or less of the vaccinated fish were infected. 相似文献
16.
Vaccination strategies with oral booster for surubim hybrid (Pseudoplatystoma corruscans × P. reticulatum) against haemorrhagic septicaemia 
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下载免费PDF全文 Gabriella do Vale Pereira Bruno Corrêa da Silva Felipe do Nascimento Vieira Walter Quadros Seiffert Thiago Tetsuo Ushizima José Luiz Pedreira Mouriño Maurício Laterça Martins 《Aquaculture Research》2015,46(8):1831-1841
This study evaluated the efficiency of differently prepared vaccines against Aeromonas hydrophila in the hybrid surubim (Pseudoplatystoma corruscans × P. reticulatum). Survival and haemato‐immunological parameters were compared between the treatments: non‐vaccinated fish (C); bacterin‐vaccinated fish (B); bacterin plus oral booster vaccinated fish (B+O); bacterin and toxoid‐vaccinated fish (B+T) and bacterin, toxoid and oral booster‐vaccinated fish (B+T+O). Fourteen‐days vaccinated fish from B+O and B+T+O were fed with an oral booster for 4 days. After 1 week, the fish were intraperitoneally challenged with 2 × 108 CFU mL?1 of A. hydrophila. Fish from the treatment B+T+O showed the lowest cumulative mortality (11.36%) 96 h after challenge, compared with other treatments (22.72–44.04%), and a relative survival of 74%. Serum immunoglobulin in B+T+O fish was higher than in other treatments. All vaccinated fish showed an increased agglutination titre when compared with non‐vaccinated fish, both before and after challenge. Fish fed with oral booster showed an increase in phagocytic percentage before and after challenge. It can be inferred that the oral booster vaccination was efficient in reducing mortality in hybrid surubim by enhancing the response against haemorrhagic septicaemia due to A. hydrophila infection. 相似文献
17.
Abstract. Turbot, Scophthalmus maximus (L.), and sea bass, Dicentrarchus labrax (L.), are susceptible to vibriosis, a septicaemia caused in France by Vibrio anguillarum 408. A bacterin produced with this strain by Rhône–Merieux has been tested orally in both species and, as a comparison, by intraperitoneal injection. During a challenge carried out 4 weeks later by intraperitoneal inoculation, the protection observed in orally vaccinated fishes was significant (72 % in sea bass and 70 % in turbot). Eleven weeks after vaccination, this protection is still significant. The mean titre of serum agglutinins was weakly increased in orally vaccinated fishes. Bacteriostatic antibodies have also been searched for in serum, but compared to agglutination, the applied technique leads to lower titres and it has not been possible to prove the presence of such antibodies after oral vaccination. Passive immunizations using serum from orally vaccinated fishes (2 months after vaccination) confers some protection against a challenge carried out by inoculation of virulent vibrio. Therefore, the production of serum factors following oral vaccination of sea bass and turbot may be involved in generalized protection of fish. 相似文献
18.
Dietary levamisole modulates the immune response and disease resistance of Asian catfish Clarias batrachus (Linnaeus) 总被引：2，自引：0，他引：2
In order to determine the immunomodulatory effect of dietary levamisole in Asian catfish (Clarias batrachus), fish were fed four different diets for 10 days: a formulated diet as control and the same diet supplemented with 50, 150 or 450 mg levamisole kg?1 feed. The serum bacterial agglutination titre against Aeromonas hydrophila as a measure of specific immunity, serum haemagglutination titre, natural haemolytic complement activity (ACH50), myeloperoxidase and lysozyme activities, total protein level and oxidative radical production by neutrophils as a measure of non‐specific immunity as well as disease resistance against A. hydrophila challenge to separate vaccinated and non‐vaccinated groups were evaluated at 0, 1, 2 and 3 weeks after last administration of levamisole. Levamisole supplement at the lowest level (50 mg kg?1) significantly enhanced oxidative radical production and serum myeloperoxidase (MPO) content immediately after 10 days of feeding, which reached peak values after 3 and 2 weeks of feeding respectively. Haemolytic complement and haemagglutination titre were significantly enhanced after 3 and 1 weeks respectively. Haemolytic complement activity and MPO activities were significantly raised to 150 mg kg?1 after 3 and 2 weeks, respectively. At the highest level of levamisole feeding (450 mg kg?1) significant decreases in superoxide production and complement activity were measured immediately after levamisole feeding, which returned to the normal level after 1 week post‐ feeding. Fish were challenged with a virulent strain of A. hydrophila at 0, 1, 2 and 3 weeks after levamisole feeding, and the cumulative per cent survival was recorded over 10 days. Feeding levamisole at 50, 150 or 450 mg kg?1 increased per cent survival in vaccinated fish immediately after levamisole feeding, and survival was significantly higher at 450 mg kg?1. There was no difference in mortality patterns in non‐vaccinated fish. The results support the use of levamisole at 50 mg kg?1 feed for 10 days as an immunostimulant in Asian catfish farming. 相似文献
19.
Onset of immunity in salmonid fry vaccinated by direct immersion in Vibrio anguillarum and Yersinia ruckeri bacterins 总被引：3，自引：0，他引：3
Abstract. The fry of several salmonid species were vaccinated by direct immersion in either Yersinia ruckeri or Vibrio anguillarum bacterin and the level of protective immunity was determined by the survival of fish after bath challenge with virulent organisms. The immersion time for effective vaccination was obtained within 5 s and protective immunity was demonstrated within 5 days at 18°C and within 10 days at 10°C. The minimum size at which maximum protective immunity occurred was between 1.0 and 2.5 g. Immunity appeared to be a function of size and not age, but differences in response among several species were indicated. In fish under 1.0 g, the level of protective immunity could be increased using a more concentrated bacterin. The results were similar with both bacterins. 相似文献
20.
注射菌苗的生殖期黄颡鱼亲鱼的外周血细胞和体液免疫变化 总被引：1，自引：0，他引：1
阐明生殖期亲鱼在注射灭活菌苗后的血细胞和体液免疫变化规律,是预防亲鱼产后虚弱患病和仔鱼染病死亡的理论基础。本研究在试验组和对照组黄颡鱼胸鳍基部分别注射0.2 mL浓度为1.0×108 CFU/mL福尔马林灭活温和气单胞菌苗（F-AS）和0.65%灭菌生理盐水,在免疫后第1、2、4、7、14、21、28、35天,从尾静脉采血,测定外周血液血细胞数量及组成比例,血细胞吞噬活性和抗体效价。结果显示：注射菌苗后白细胞数量和淋巴细胞的白细胞分类计数逐渐增加,显著高于对照;中性粒细胞、单核细胞的白细胞分类计数以及吞噬百分比（PP）、吞噬指数（PI）在4～7 d显著高于对照;抗体效价逐渐升高且在第14天达到最高,随后逐渐下降;而红细胞数和血栓细胞的白细胞分类计数逐渐下降,显著低于对照组。结果表明：单次注射菌苗后亲鱼可以产生良好的免疫应答,1～7 d以增强单核细胞和中性粒细胞等非特异细胞免疫为主;7～35 d淋巴细胞增值,并大量释放抗体且在第21天淋巴细胞出现极显著升高,此后虽逐渐下降但仍显著高于对照,抗体维持在较高水平上的时间为1周左右,建议在黄颡鱼催产前10～14 d进行单次菌苗注射以增强亲鱼免疫力。 相似文献