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1.
3种免疫途径对嗜水气单胞菌灭活疫苗保护作用的影响 总被引：2，自引：0，他引：2
应用间接ELISA方法,研究经嗜水气单胞菌疫苗浸泡、口服、注射免疫后鳜皮肤、肠道黏液和血清中抗体的变化关系,揭示鳜对3种免疫途径的免疫应答效果。结果显示:黏液抗体产生快(7 d达到峰值),但持续时间短,抗体水平低(213);血清抗体产生慢(28 d方达峰值),但持续时间长,抗体水平高(225)。不同方式免疫鳜后均产生远高于对照组的抗体滴度(P<0.05)。注射组产生的抗体水平高(225),免疫保护率最理想(70.6%);口服组相对另两个实验组,其抗体峰值(215)和免疫保护率(41.2%)均较低;浸泡组在皮肤黏液产生水平和溶菌酶含量方面,产生较好的免疫效应,分别为213和178μg/mL,但相对保护率仅为47.1%,低于注射组的70.6%。浸泡和口服组可诱发局部黏膜免疫,在抗体动态变化方面表现出相似的规律且峰值时间早于注射组。初步推断浸泡和口服可以直接刺激鱼体黏膜组织产生局部的特异性抗体。 相似文献
2.
《广东海洋大学学报》2016,(4)
从患烂尾病眼斑拟石首鱼(Sciaenops ocellatus)的溃疡处分离到一株优势菌So GZ1401,回归感染试验证实该菌是引起烂尾病的病原菌,对眼斑拟石首鱼的半致死量(LD50)为8.6×103 CFU·g~(-1);结合其形态学和生理生化特征及HSP60测序分析,鉴定为哈维氏弧菌(Vibrio harveyi);利用杯碟法研究其胞外产物性质,结果表明:其胞外产物具有淀粉酶、明胶酶、酪蛋白酶、卵磷脂酶、脂酶活性以及溶血活性,但无脲酶活性。药敏试验表明,该菌株对氟哌酸、恩诺沙星、氟苯尼考和复方新诺明等抗菌药有较高的敏感性。制备哈维氏弧菌灭活疫苗,采用注射和浸泡2种途径免疫眼斑拟石首鱼,测定受免疫鱼的血清抗体效价和疫苗对眼斑拟石首鱼的免疫保护率。结果显示,免疫组血清中的抗体效价水平均高于对照组,差异有统计学意义(P0.05),注射组和浸泡组的抗体效价最高值分别达718.4和128,哈维氏弧菌攻毒后免疫保护率分别为75.0%~86.7%和46.4%~56.7%。哈维氏弧菌灭活疫苗对眼斑拟石首鱼具有较好的免疫保护性。 相似文献
3.
制备无乳链球菌(Streptococcus agalactiae)临床分离株ZJ02的灭活疫苗,采用注射和浸泡2种方法,研究疫苗对吉富罗非鱼(Oreochromis niloticus)的免疫保护效果和血清抗体效价。注射组以4个不同浓度的白油佐剂灭活疫苗免疫罗非鱼,浸泡组以相同免疫剂量的灭活疫苗分别与消旋山莨菪碱、脂肪醇聚氧乙烯醚和氯化钠3种佐剂免疫罗非鱼。结果显示,无乳链球菌疫苗对罗非鱼具有较强的免疫原性,与对照组相比,免疫组血清中的抗体效价水平均显著性提高(p<0.05),注射组和浸泡组的抗体效价最高分别达到1∶256和1∶128;注射组中不同剂量的疫苗免疫组免疫保护率较高,高浓度组可达到95%,浸泡组中使用3种佐剂的免疫组免疫保护率分别为59%、64%、55%。表明无乳链球菌灭活疫苗对罗非鱼具有很好的免疫保护性,免疫剂量在5×108~4×109cfu/mL之间对注射免疫效果有显著性影响,消旋山莨菪碱、脂肪醇聚氧乙烯醚和氯化钠对浸泡免疫效果具有不同的免疫保护率。 相似文献
4.
实时荧光定量PCR检测鳜IgM mRNA标准品质粒的构建 总被引：1，自引：0，他引：1
利用SYBR Green I荧光定量PCR技术,建立鳜IgM-DNA定量标准品的制备方法。从浸泡免疫后的鳜头肾中提取总RNA逆转录合成cDNA,对目的片段进行PCR扩增、电泳纯化、T-A克隆及测序鉴定。将所得预期质粒梯度稀释后构建标准曲线并进行融解曲线分析。结果表明,当标准品的浓度在3.29×102～3.29×108拷贝/μL时,模板浓度与循环阈值(Ct)间的线性关系良好,相关系数r2达到0.999;融解曲线分析显示具特异的单个峰,表明扩增产物特异性非常好。此法制备的重组质粒标准品可用于对鳜IgM基因的转录水平进行测定。 相似文献
5.
用鲶爱德华氏菌兔抗血清作为一抗,碱性磷酸酶(AP)标记的羊抗兔IgG作为酶标二抗,建立黄颡鱼"红头病"病原菌—鲶爱德华氏菌的间接酶联免疫(ELISA)快速检测法,并优化检测条件。抗原最佳包被浓度为107/mL,一抗工作的最佳稀释度为1∶211,病原菌的检测灵敏度为105/mL,交叉反应实验证明该方法特异性强,与迟钝爱德华氏菌、弧菌等13种标准菌株无交叉。应用上述技术对人工感染发病鱼中分离的优势菌进行检测,结果表明阳性检出率为80%;对自然发病黄颡鱼体内分离获得的20株优势菌检测结果表明,12株菌为鲶爱德华氏菌。 相似文献
6.
以黄颡鱼"红头病"致病菌——鮰爱德华菌(Edwardsiella ictaluri)A86作为实验菌株,通过十二烷基肌氨酸钠(Sarkosyl)抽提并结合超速离心的方法提取主要外膜蛋白(OMPs),SDS-PAGE图谱分析结果显示,OMPs主要有6条条带,相对分子质量分别为45 000、42 000、41 000、36 000、30 000和22 000。用A86 OMPs免疫新西兰大白兔,获得多克隆抗体22 mL,抗体效价为1∶20 480,抗体按照1∶2 560稀释时具有较高的特异性,仅与迟缓爱德华菌(E.tarda)、海豚链球菌(Streptococcus iniae)和创伤弧菌(Vibrio vulnificus)存在微弱的交叉反应,与鮰爱德华菌参考株及不同地区的分离株呈现特异性结合。多克隆抗体与OMPs的免疫印迹结果显示,主要有2条明显的免疫反应发色带,相对分子质量分别为36 000和30 000,且36 000蛋白染色最为明显。多克隆抗体与包括菌株A86在内的分离自南北方不同地区的10株鮰爱德华菌全菌免疫印迹试验发现,所有受检菌株反应结果相同,主要有4条明显的反应条带,相对分子质量分别为106 000、54 000、36 000和30 000,其中36 000和30 000两条蛋白带染色最深。因此,认为从黄颡鱼体内分离的鮰爱德华菌OMPs中36 000和30 000蛋白具有很好的免疫原性,36 000蛋白尤甚。 相似文献
7.
寡糖对三黄鸡生产性能及免疫机能的影响 总被引：2，自引：0，他引：2
用96只1日龄广东三黄鸡,研究了饲料添加0.2%或0.3%的寡糖对肉鸡生产性能与免疫机能的影响。结果表明:饲料中添加0.2%的寡糖能显著提高11~30日龄肉鸡的增重、免疫器官指数及新城疫H I抗体滴度(P<0.05),血液白细胞吞噬率和血清IgG含量亦高于对照组,但组间差异不显著;添加0.2%或0.3%的寡糖能明显改善全期的饲料转化率(P<0.05)。 相似文献
8.
以红笛鲷Lutjanus sanguineus弧菌病的主要病原菌——溶藻弧菌Vibrio alginolyticus为抗原制备兔抗血清,以辣根过氧化酶标记的羊抗兔血清为酶标二抗,建立了酶联免疫吸附试验检测技术。通过棋盘滴定法测定,得出溶藻弧菌菌悬液的最佳工作浓度为5×108mL-1,兔抗溶藻弧菌血清的最佳稀释度为1∶16000,酶标羊抗兔抗体的最佳稀释度为1∶2000。应用本实验建立的间接双抗体夹心法对红笛鲷进行现场检测,患病红笛鲷特异性抗体的检出率为70%,外观健康红笛鲷的检出率为20%。结果表明,本实验所建立的酶联免疫吸附试验(ELISA)检测法可以用于红笛鲷弧菌病的血清流行病学研究,而且可以检测出隐性感染状态下的红笛鲷。 相似文献
9.
吉富罗非鱼最适生长水温研究 总被引：3，自引：0，他引：3
为研究不同养殖水温对吉富罗非鱼摄食和生长的影响,了解吉富罗非鱼的最适生长水温,实验设计28℃、30℃、32℃、34℃4个不同温度梯度,周期45 d。结果表明:不同水温环境下吉富罗非鱼表现出不同的生长特性,养殖在30℃水体中的罗非鱼生长速度明显高于其他温度组;水温对吉富罗非鱼的特定生长率、绝对增重率、饲料转化率、摄食率有显著影响(P<0.05),之间的相关关系可以用二次回归曲线来描述,特定生长率(SGR)与水温的关系式为rSGR=-19.255+1.3794T-0.0235T2,摄食率(FR)与水温的关系式为rFR=-22.175+1.5901T-0.026 6 T2,饲料转化率(FCR)与水温的关系式为rFCR=34.041-2.2256T+0.0382 T2;吉富罗非鱼的最适生长水温为29.3℃,最大摄食率的水温为29.9℃,最高食物转化率水温为29.1℃。 相似文献
10.
溶藻弧菌疫苗对凡纳滨对虾免疫功能的影响 总被引：3，自引：0，他引：3
应用溶藻弧菌疫苗饲喂凡纳滨对虾,测定了弧菌疫苗对凡纳滨对虾免疫指标的影响以及 疫苗的免疫保护作用。结果发现,投喂弧菌疫苗后,溶菌活力在12～20d内迅速增强,在20d时 活力达到最高,VF1组高于VF2和VF3组,但VF2和VF3之间差异不显著;凝集效价在投喂后的 第8天开始升高,在14d时达到最高,实验组的凝集活力均高于对照组,但实验组之间无明显的差 异;而超氧化物歧化酶(SOD)活力和溶血活力变化不明显。以每克饲料含疫苗细胞1.0×109、 1.0×108和1.0×107个的浓度投喂,对凡纳滨对虾的免疫保护率分别为66.7%、46.7%和13.3%。 可见弧菌疫苗对凡纳滨对虾的免疫保护率随疫苗浓度的增大而升高。 相似文献
11.
Bacterin was prepared by formalin-inactivating the virulent strain of Vibrio hollisae isolated from diseased Seriola dumerili （amberjack） suffering from vibriosis. Healthy S. dumeriIi were vaccinated by respective procedures of intramuscular injection, immersion, and orally administration. Results of the three different vaccinations were compared. Blood was drawn from the vaccinated fish every 7 days, and the antibody titers and lysozyme activities of the sera were determined. The antibody titer of injected fish was 1：40 at 7 d, and reached its peak of 1：320 at 28 d, while the fish vaccinated by immersion and orally administration exhibited weak antibody responses, the antibody titres of 〈1：10, 1：20, 1：160 were observed at 7 d, 14 d, 35 d respectively. Compared with the control, the vaccinated fish exhibited significantly higher lysozyme activities （P〈0.05）. Upon challenge with virulent strain, the relative percent survival （RPS） of injected, immersed and oral administrated fish were 75%, 45%, and 40% respectively, and the injected fish showed significantly higher RPS than immersed and oral administrated fish. The results suggested that vaccination of S. dumerili by the injection would be the best strategy to prevent the vibriosis in S. dumerili farm. 相似文献
12.
SERUM IMMUNOGLOBULIN OF THE MANDARIN FISH,SINIPERCA CHUATSI WITH DEVELOPMENT OF POLYCLONAL ANTIBODY 总被引：2，自引：0，他引：2
Serum immunoglobulin from the mandarin fish,or the so-called Chinese perch,Siniperca chuatsi(Basilewsky),was successfully purified using affinity chromatography.Heavy and light chains were detected on electrophoresis gel,with molecular weights being estimated at 72 and 29 kDa,respectively.The tetrameric IgM of S.chuatsi was calculated to be 808 kDa.The rabbit polyclonal antisera against the purifed immunoglobulin were developed and tested by Westem blot analysis.The antisera reacted strongly with the heavy chains of S.chuatsi immunoglobulin.Humoral immune responses of the mandarin fish can then be examined using the developed polyclonal antibody. 相似文献
13.
Bacterin was prepared by formalin-inactivating the virulent strain of Vibrio hollisae isolated from diseased Seriola dumerili (amberjack) suffering from vibriosis. Healthy S. dumerili were vaccinated by respective procedures of intramuscular injection, immersion, and orally administration. Results of the three different vaccinations were compared. Blood was drawn from the vaccinated fish every 7 days, and the antibody titers and lysozyme activities of the sera were determined. The antibody titer of injected fish was 1:40 at 7 d, and reached its peak of 1:320 at 28 d, while the fish vaccinated by immersion and orally administration exhibited weak antibody responses, the antibody titres of <1:10, 1:20, 1:160 were observed at 7 d, 14 d, 35 d respectively. Compared with the control, the vaccinated fish exhibited significantly higher lysozyme activities (P<0.05). Upon challenge with virulent strain, the relative percent survival (RPS) of injected, immersed and oral administrated fish were 75%, 45%, and 40% respectively, and the injected fish showed significantly higher RPS than immersed and oral administrated fish. The results suggested that vaccination of S. dumerili by the injection would be the best strategy to prevent the vibriosis in S. dumerili farm. 相似文献
14.
Microorganisms living in polar zones play an important part as the potential source of organic activity materials with low temperature characteristics in the bio-technological applications. A psychrotrophic bacterium (strain Ar/w/b/75°/10/5) , producing cellulose at low temperatures during late-exponential and early-stationary phases of cell growth, was isolated from sea ice-covered surface water in Chuckchi Sea, Arctic. This bacterium, with rod cells, was Gram-negative, slightly halophilic. Colony growing on agar plate was in black. Optimum growth temperature was 15℃. No cell growth was observed at 351 or above. Optimum salt concentration for cell growth was between 2 and 3 % of sodium chloride in media. Maximal cellulase activity was detected at a temperature of 35℃ and pH8. Cellulase was irreversibly inactivated when incubated at 55℃ within 30 min. Enzyme can be kept stable at the temperature no higher than 25℃. Of special interest was that this bacterium produced various extracellular enzymes i 相似文献
15.
Activities of hexokinase(HK), pyrnvate kinase(PK)and levels of HSPT0 were measured to evaluate the response of Litopenaeus vannamei to rapid temperature changes under controlled laboratory conditions. Shrimps were subjected to a quick temperature change from 27℃ to 17℃ for the summer case(Cold temperature treatment), or from 17℃ to 27℃ for the winter case(Warm temperature treatment). After 0. 5, 1, 3, 6, 12, 24, 48, and 72 h of exposure time, shrimps were sampled and prepared for further analysis. The results showed that the effect of acute temperature changes on activities of HK was significant. Patterns of variations of the two glycolytic enzymes suggested that enzymes in the glycolysis cycle could adjust their activities to meet the acute temperature change. The HSP70 level increased in both cold and warm temperature treatments, suggesting that the rapid temperature changes activated the process of body's self-protection. But the difference in expression peak of HSP70 might be related to the different body size and the higher thermal sensitivity to temperature increase than to temperature decrease of L. Vannamei. 相似文献
16.
Activities of hexokinase(HK),pyruvate kinase(PK) and levels of HSP70 were measured to evaluate the response of Litopenaeus vannamei to rapid temperature changes under controlled laboratory conditions.Shrimps were subjected to a quick temperature change from 27℃ to 17℃ for the summer case(Cold temperature treatment),or from 17℃ to 27℃ for the winter case(Warm temperature treatment).After 0.5,1,3,6,12,24,48,and 72 h of exposure time,shrimps were sampled and prepared for further analysis.The results showed that the effect of acute temperature changes on activities of HK was significant.Patterns of variations of the two glycolytic enzymes suggested that enzymes in the glycolysis cycle could adjust their activities to meet the acute temperature change.The HSP70 level increased in both cold and warm temperature treatments,suggesting that the rapid temperature changes activated the process of body's self-protection.But the difference in expression peak of HSP70 might be related to the different body size and the higher thermal sensitivity to temperature increase than to temperature decrease of L.vannamei. 相似文献
17.
干热岩地热开发中的钻井、储层压裂及热交换等环节均涉及高温岩石冷却的问题,为揭示其中岩石损伤演化规律,基于巴西劈裂试验和声发射技术,研究了不同高温及冷却方式对花岗岩抗拉性质的影响。结果表明:①25~600℃下花岗岩抗拉强度随温度升高而下降,遇水冷却使抗拉强度进-步下降并使其开始大幅下降的温度阈值提前到200,500℃后抗拉强度对遇水冷却更敏感。②荷载达到峰值,声发射累计振铃计数突增,岩样内形成断裂区;受遇水冷却影响,岩样的振铃计数峰值和能量峰值有所下降,间接反映岩石内裂纹更发育,200~300℃时降幅均较大,300℃时和500℃后花岗岩对热处理方式较敏感。③花岗岩破裂面随温度升高由平整向粗糙曲折变化,由脆性向延性转变,遇水冷却促进岩石破裂并促使脆性向延性转变的温度区间提前。研究结果为地热开采中高温岩石的稳定性评价提供理论参考。 相似文献
18.
LIU Chune LIN Hong CAO Limin JIANG Jie 《中国海洋大学学报(英文版)》2005,4(3):262-266
A two-step zero-length cross-linking procedure using active esters was successfully adopted for conjugating enrofloxacin （EF） to human serum albumin （HSA）. The derived conjugate was characterized by UV spectrum and then used for immunization of BALB/C mice. In enzyme-linked immunosorbent assay （ELISA） and competitive inhibition ELISA experiments, the derived antiserum exhibited high antibody titer （greater than 1 ： 250 000） as well as varied cross-reactivity （from 97.8% to 161.7%） to three analogs of EF belonging to fluoroquinolones family. But over the concentration range studied, no significant cross-reactivity was observed to other group of antibiotics （chloramphenicol, oxytetracycline, sulphamethoxazole and nysfungin）. It was confirmed that the synthesized immunogen was highly antigenic and elicited specific antibody responses in BALB/C mice against EF. 相似文献