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1.
Protective immunity against enteric septicaemia of catfish (ESC) following immunization with Edwardsiella ictaluri bacterins and exposure to live E . ictaluri was investigated. Mean cumulative percentage survival was significantly higher ( P 0.05) in controlled live vaccinates (100%) than in immersion and oral bacterin vaccinates (68.3% and 50.0%, respectively). Bactericidal activity against E . ictaluri by peritoneal macrophages from controlled live vaccinates (85.9%) was significantly greater ( P 0.05) than bactericidal activity of macrophages from immersion bacterin vaccinates (71.4%) or non-vaccinates (68.1%). No significant ( P > 0.05) difference was found in the bactericidal activity of macrophages from oral bacterin vaccinates and macrophages from controlled live vaccinates. The E . ictaluri -specific antibody response of controlled live (0.08 OD) and immersion bacterin vaccinates (0.11 OD) was significantly higher ( P 0.05) than that of oral bacterin vaccinates and non-vaccinates (0.01 OD) 15 days post-vaccination. A significantly higher antibody response was seen in controlled live vaccinates (0.17 OD), when compared to other vaccinates or non-vaccinates 33 days after vaccination. Neither immersion nor oral bacterins protected the vaccinates against ESC. Controlled live E . ictaluri immunization of channel catfish resulted in production of specific antibodies, increased macrophage bactericidal activity and protection against ESC.  相似文献   

2.
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   

3.
Abstract. Mutants of Aeromonas salmonicida strains lacking either the A-protein, O-antigen or both of these major surface antigens were tested in rainbow trout, Oncorhynchus mykiss (Walbaum), for their suitability as live vaccines (LV). All of these mutants were shown to be attenuated, as fish receiving ∼5 × 107 of the respective strains showed no clinical signs of furunculosis. Immersion vaccination of fish in 5 × 107 cfu ml-1 of these strains with an identical immersion dose 14 days later resulted in significant protection by all strains from challenge with a heterologous virulent strain of A. salmonicida 5 weeks later. The levels of protection conferred were all greater than or equal to that provided by an injected bacterin using the same vaccination schedule. With one exception, all LV strains that still possessed a functional O-antigen provided protective indices (PI) four- to seven-fold greater than the PI for the fish injected with bacterin. When antibody responses of vaccinated fish were compared, it was found that only vaccination by bacterin gave rise to a measurable agglutinating litre. Western immunoblots using the immune fish sera failed to reveal any major differences in antigen recognition in fish that received any of the vaccines tested. These data suggest that the immune response generated by the use of live vaccine strains is different from that generated by a bacterin, and that these useful mutations may be incorporated into existing furunculosis LVs for further attenuation.  相似文献   

4.
Passive immunization of tilapia, Oreochromis niloticus, was conducted to determine whether anti- Streptococcus iniae whole sera (ASI), heat inactivated anti- S . iniae whole sera (HIASI) and normal whole sera (NWS) were protective when intraperitoneally (i.p.) injected into tilapia. The ASI was produced in tilapia actively immunized (challenged) with virulent S. iniae by i.p. injection. An antibody response against S. iniae was demonstrated by enzyme linked immunosorbent assay (ELISA) and 18% of the immunized fish died because of the S. iniae infection. The actively immunized tilapia demonstrated a secondary antibody response and immunity to S. iniae after challenge with S. iniae by i.p. injection. Survival was 100% in the actively immunized fish. The NWS was obtained from tilapia free of ASI antibody and susceptible to S. iniae infection (40% mortality). In two separate experiments, significantly higher mortality was noted in tilapia passively immunized with NWS (33 and 53%) and phosphate buffered saline (PBS) (30 and 60%), in comparison with mortalities of 0 and 10% or 3.3 and 6.7% in the fish passively immunized with ASI or HIASI 14 days after S . iniae infection by i.p. injection ( P  = 0.0003 and 0.0023). Results suggest that immunity provided by ASI and HIASI was because of antibody against S. iniae . Inactivation of complement in the HIASI treatment further suggests that ASI antibody plays a primary role in immunity against S. iniae infection.  相似文献   

5.
为了解日本鳗鲡(Anguilla japonica)受嗜水气单胞菌(Aeromonas hydrophila)感染后血清酶活力的变化规律,将日本鳗鲡分为感染组和对照组,每组30尾,腹腔分别注射0.1 mL浓度为3×106CFU/mL的菌液和等量灭菌生理盐水。在注射后0、1、5、9、13、17、21、25、29、33 d从两组各取3尾日本鳗鲡,尾静脉取血,测定其酸性磷酸酶(ACP)、超氧化物歧化酶(SOD)、一氧化氮合酶(NOS)活力的变化规律。结果显示:注射嗜水气单胞菌一段时间后,实验组鳗鲡的ACP和SOD的活力都显著高于对照组,且都在21 d时达到最高值,随后逐渐下降。结果表明:嗜水气单胞菌感染后,日本鳗鲡能够通过提高血清中ACP和SOD活力来提高机体对嗜水气单胞菌的免疫力,而对一氧化氮合酶的活力没有显著影响。  相似文献   

6.
制备哈氏弧菌SpGY020601外膜蛋白复合物(OMPC)和溶藻弧菌EpGS021001脂多糖(LPS)偶联疫苗,设置偶联疫苗组、偶联疫苗 ISA763佐剂组、偶联疫苗 β-葡聚糖佐剂(DEAE)组以及生理盐水对照组,并以相同程序免疫卵型鲳鲹。溶菌酶活性检测、微量凝集反应试验结果显示:各疫苗组血清中的溶菌酶含量无显著差异(P>0.05),但均远高于生理盐水对照组(P<0.05);ISA763佐剂组抗哈氏弧菌SpGY020601抗体滴度较其他组出现的早,抗溶藻弧菌EpGS021001抗体滴度较其他组持续时间长;β-葡聚糖佐剂组次之,其次是无佐剂组,对照组始终未检测到抗体。各组分为腹腔注射攻毒与浸泡攻毒,对EpGS021001的攻击,ISA763佐剂组相对保护率分别为90%和70%,高于β-葡聚糖佐剂组的85%和55%,以及无佐剂组的80%和55%;对SpGY020601的攻击,ISA763佐剂组相对保护率分别为95%和80%,高于或相当于β-葡聚糖佐剂组的95%和75%,以及无佐剂组的90%和75%。  相似文献   

7.
Abstract. Potassium aluminium sulphate (alum) was tested for use as an adjuvant in an Aeromonas salmonicida bacterin injected into Atlantic salmon, Salmo salar L. Potential side-effects of visceral adhesions, diminished growth and toxicity, as well as adjuvant effects, were examined. Both alum and the bacterin alone induced inflammation and adhesion formation when injected into the peritoneal cavity. When bacterin and alum were combined, these effects were compounded. Growth was not affected by any of the injected treatments. The acute toxicity of the bacterin was reduced from 33.7 to 0.3% mortality by the addition of alum. Alum appeared to enhance protection against challenge, but there were no statistically significant differences in protection between injecting bacterin alone or injecting bacterin with alum.  相似文献   

8.
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   

9.
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   

10.
Goldfish, Carassius auratus (L.), were intraperitoneally injected with two doses of Ichthyophthirius multifiliis (each containing 2.3 × 106 live tomites) in saline, 30 days apart. The control fish were injected with saline only. Two weeks after the last vaccination, fish of both groups were intradermally injected with 2.5 × 105 live tomites for skin tests. The skin at the antigen-injected site of the vaccinated fish increased in thickness from 8 h onwards and reached a peak at 24 h. Histological study showed a heavy infiltration of mononuclear leucocytes at the antigen-injected site. In contrast, no such changes were encountered in control fish. The migration area of pronephros cells from the vaccinated fish was significantly inhibited, in vitro, compared to controls with the presence of antigens. These findings suggest that the tomite-vaccinated fish produced a cell-mediated immune response. However, vaccinated fish also exhibited significantly higher titres of immobilizing antibodies in their sera and mucus than the controls. Therefore, it is likely that both humoral and cell-mediated immune responses have to work closely together to eliminate the infectious tomites of I. multifiliis which succeed in penetrating the fish epithelial tissues.  相似文献   

11.
Largemouth bass Micropterus salmoides were immunized with four different N. seriolae strains—two α-glucosidase-positive (961113, KU040801) and two α-glucosidase-negative (94260, OTTS) strains—along with Freund’s incomplete adjuvant. After primary immunization (week 0), a booster was administered at weeks 4 and 8. Nonspecific immune responses to multiple immunizations with the different N. seriolae strains were determined based on serum lysozyme activity and nitroblue tetrazolium (NBT)-positive cells in peripheral blood. The serum lysozyme activity and NBT-positive cells in peripheral blood were not significantly increased even after the two booster immunizations. Specific antibody responses against N. seriolae cells were investigated by enzyme-linked immunosorbent assay. At 4 weeks after immunization, all groups immunized with N. seriolae antigens showed significant increases in their specific antibody levels. The sera from fish immunized with different N. seriolae strains exhibited reactivity with N. seriolae sonicated antigens of 28, 30, 36 and 84 kDa by western blot analysis. After two boosters, fish were challenged with live N. seriolae to assess the vaccine’s efficacy; however, multiple injections of the N. seriolae strains did not reduce mortality, irrespective of the bacterin.  相似文献   

12.
以初体重为(7.20±1.38)g的凡纳滨对虾(Litopenaeus vannamei)为研究对象,在室内养殖箱进行3周的养殖实验和2周的哈维氏弧菌(Vibrio harveyi)人工感染实验,其中对照组每日投喂普通商品饲料,实验组每日投喂在普通商品饲料中添加地衣芽孢杆菌(Bacillus licheniformis)1.0×10~8 CFU/m L配制成的实验饲料。目的是研究饲料中添加益生菌对凡纳滨对虾肠道菌群、Toll受体及溶菌酶基因表达量和抗哈维氏弧菌能力的影响。实验结果表明,在饲料中添加地衣芽孢杆菌可显著提高对虾抗哈维氏弧菌感染的能力,其相对免疫保护率为22.22%。与对照组相比,实验组可显著降低对虾肠道内弧菌数量(P0.05)。感染哈维氏弧菌后,实验组溶菌酶(lysozyme,LZM)m RNA的相对表达量在12 h、18 h、24 h、36 h、48 h、72 h均显著高于对照组(P0.05)。实验组感染哈维氏弧菌后在6 h、12 h、18 h、24 h、36 h、48 h、72 h、7 d的Toll受体(Toll receptor)m RNA的相对表达量均显著高于对照组(P0.05)。实验结果提示:饲料中添加地衣芽孢杆菌可有效提高对虾抗哈维氏弧菌感染的能力,这种能力的提高可能是通过降低对虾肠道内的弧菌量,并提高抗病相关基因的表达量实现的。  相似文献   

13.
Abstract. A formalized bacterin of Vibrio anguillarum , with and without alum adjuvant, was administered intraperitoneally to rainbow trout under farm conditions. Compared with the normal bacterin the adjuvanted preparation caused pathological changes indicative of chronic peritonitis, substantial early mortalities in 3 cm fish, and to a lesser extent in 20 cm fish, and depressed growth rate. No additional degree of protection against artificial challenge was obtained from its use.  相似文献   

14.
The protection of cultured sole, Solea senegalensis, against Vibrio harveyi and Photobacterium damselae subsp. piscicida was evaluated following the use of a divalent vaccine prepared with formalized whole cells and extracellular products of virulent strains of both pathogenic microorganisms and administered by the immersion route. Two prolonged immersions of 5-10 g fish in the divalent bacterin at a 1-month interval gave high levels of protection similar to those obtained when the respective monovalent vaccines were administered by the intraperitoneal route [relative percentage of survival (RPS) values >70%], which indicates that the former procedure can be a useful strategy with small fish. The high protection afforded by the divalent vaccine in sole lasted for 4 months after which the RPS values against both pathogens decreased significantly.  相似文献   

15.
Bacterial subcellular components and probiotics were successful for the stimulation of immunity and the prevention of Vibrio harveyi infections in rainbow trout, Oncorhynchus mykiss (Walbaum). Rainbow trout were immunized with whole inactivated cells of V. harveyi to obtain polyclonal antibodies against specific antigens. Western blotting showed a unique reactive band (∼93 kDa) between serum and bacterial proteins from outer membrane proteins (OMP) and extracellular products (ECP). Probiotics were selected according to their capability to inhibit V. harveyi . Two of these bacteria, i.e. A3-47 and A3-51, showed cross-reactivity with V. harveyi antiserum. Their OMPs and ECPs were reactive with V. harveyi antiserum in bands of ∼93 kDa for A3-51 and higher for A3-47. In vivo tests determined that fish fed with A3-51 produced cross-reactive antibodies against V. harveyi and also, the survival of these fish infected with V. harveyi was high, being similar to the level achieved with vaccinated fish. Thus, the probiotics, when administered as live preparations, were capable of producing cross-reactive antibody against specific bacterial pathogens.  相似文献   

16.
采用十二烷基肌氨酸钠法提取哈氏弧菌(Vibrio harveyi)SpGY020601株的外膜蛋白(OMPC),采用酚水法提取溶藻弧菌(Vibrio alginolyticus)EpGS021001株的脂多糖(LPS).通过碳化二亚铵(EDC)介导的缩合反应,将哈氏弧菌的OMPC与溶藻弧菌的LPS偶联.OMPC-LPS偶联物、未偶联的哈氏弧菌OMPC、溶藻弧菌LPS、哈氏弧菌OMPC和溶藻弧菌LPS的简单混合物以及生理盐水按相同程序免疫卵形鲳鲹(Trachinotus cvatus).检测血清溶菌酶活性、血清抗体微量凝集反应结果显示,卵型鲳鲹经OMPC-LPS偶联物、OMPC、LPS以及二者的简单混合物免疫后,各免疫组间血清溶菌酶活性没有显著差异(P＞0.05),但均显著高于生理盐水对照组(P＜0.05);OMPC-LPS偶联物免疫组的血清抗溶藻弧菌EpGS021001抗体效价较单纯溶藻弧菌LPS免疫组、简单混合物免疫组出现得早,抗体滴度高;与此类似,OMPC-LPS偶联组中抗哈氏弧菌SpGY020601的血清抗体效价较单纯哈氏弧菌OMPC组、简单混合组出现得早,抗体滴度高,且持续时间长;对EpGS021001、SpGY020601的攻击,OMPC-LPS偶联物免疫组的保护率分别为85%和95%,高于单纯溶藻弧菌LPS免疫组的70%、单纯哈氏弧菌OMPC免疫组的75%,也高于简单混合物免疫组的80%和82.5%.  相似文献   

17.
Abstract. The histological responses of ayu, Plecoglossus altivelis , given an intramuscular injection of a formalin-killed bacterin of Vibrio anguillarum are described. Lesions at the site of injection showed muscular necrosis and infiltration of inflammatory cells by the second day after injection, and production of granulation tissue from the fifth to the fourteenth day. Protective responses against vibriosis were studied histopathologically in ayu that were vaccinated by intramuscular injection with formalin-killed Vibrio bacterin and by immersion in sonicated Vibrio bacterin, and challenged by a subcutaneous injection with viable Vibrio on the fourteenth day after the vaccination.
Efficacy of both methods was confirmed by the survival of vaccinated fish after the challenge. There was slight bacterial multiplication in the fish, and bacterial phagocytosis by infiltrated neutrophils and tissue necrosis in the injected area on the third day after the challenge. In contrast, non-vaccinated fish died within 24h of challenge, with extensive bacterial multiplication and tissue necrosis in the injected area and visceral organs.  相似文献   

18.
Four bacterial isolates from farmed gilthead sea bream, Sparus aurata, included in a previous study as members of the Vibrionaceae and Pseudomonodaceae and the genus Micrococcus, have been evaluated for their adhesive ability to skin and intestinal mucus of farmed Senegalese sole, Solea senegalensis, and their antagonistic effect on Vibrio harveyi, a pathogen of sole. These isolates showed higher adhesion to sole mucus than the pathogenic strains of V. harveyi assayed. Only two of the isolates showed antagonistic activity to V. harveyi. Interactions of the four isolates with V. harveyi in respect of adhesion to skin and intestinal mucus under exclusion, competition and displacement conditions were studied. Three isolates were able to reduce the attachment to skin and intestinal sole mucus of a pathogenic strain of V. harveyi under displacement and exclusion conditions, but not under competition conditions. The in vivo probiotic potential of isolate Pdp11 was assessed by oral administration followed by challenge with the pathogenic V. harveyi strain Lg14/00. A group of 50 Senegalese sole received a commercial diet supplemented with 10(8) cfu g(-1) of lyophilized Lg14/00 for 15 days. A second group of fish received a non-supplemented commercial diet. After challenge the mortality of the fish receiving the diet supplemented with the potential probiotic isolate was significantly lower than that in the fish receiving the non-supplemented commercial diet. This study has shown that the ability to interfere with attachment of pathogens, as well as the adhesion to host surfaces, are suitable criteria for selection of candidate probiotics for use in the culture of Senegalese sole.  相似文献   

19.
为研究三联特异性卵黄抗体对哈维氏弧菌、溶藻弧菌与副溶血弧菌的体外抑菌效果,通过制备哈维氏弧菌、溶藻弧菌与副溶血弧菌三联灭活疫苗,免疫蛋鸡。收集鸡蛋制备卵黄抗体并检测其效价、纯度和特异性。通过免疫荧光、扫描电镜和体外抑菌实验初步探究其制备的三联特异性卵黄抗体对三种弧菌的体外抑菌效果。结果显示,ELISA检测特异性卵黄抗体效价高达1:51200,并维持5周;SDS-PAGE对分离纯化的卵黄抗体分析显示,随着纯化步骤的进行,卵黄抗体中的蛋白杂带逐渐减少,纯度变高。免疫荧光和免疫印迹实验表明,特异性卵黄抗体与抗原的结合具有较高的特异性。扫描电镜进一步观察发现,相比非特异性卵黄抗体,特异性卵黄抗体处理过的弧菌,菌体互相黏附在一起,菌体细胞表面粗糙,细胞壁破损。在体外抑菌实验中,特异性卵黄抗体对弧菌具有明显的抑制效果,且抑制作用呈现剂量依赖性。综上所述,本研究制备的三联特异性卵黄抗体能够与哈维氏弧菌、溶藻弧菌和副溶血弧菌发生特异性结合,通过体外实验发现,卵黄抗体通过凝集和黏附,破坏菌体细胞的完整性,从而发挥抑菌作用。  相似文献   

20.
研究注射葡萄糖对吉富罗非鱼(Oreochromis miloticus)生化指标、胰岛素和糖酵解酶的影响。选取体质量约80 g的吉富罗非鱼150尾,随机分为2个实验组,对照组腹腔注射0.7%的无菌生理盐水,处理组按照30 mg/100 g(体质量)的剂量腹腔注射葡萄糖。结果表明:(1)吉富罗非鱼在注射葡萄糖后1 h血糖达到最高水平,而后显著下降(P<0.05),3 h后降低到正常水平。注射葡萄糖可显著提高胆固醇的含量(P<0.05),并显著降低谷草转氨酶水平(P<0.05),但对血浆蛋白、甘油三酯和谷丙转氨酶水平没有显著影响(P>0.05)。(2)肝糖原在葡萄糖注射后的6 h达到最高水平,而后显著下降(P<0.05),肌糖原没有发生变化。(3)血浆和肌肉中的胰岛素均在葡萄糖注射后的3 h达到最高水平,而后显著下降(P<0.05),而肝中的胰岛素水平没有发生变化。(4)丙酮酸激酶的活力在葡萄糖注射后6h达到最高水平(P<0.05),己糖激酶的活力在注射葡萄糖后没有发生变化。结果表明,注射葡萄糖可显著提高吉富罗非鱼血糖水平,且维持时间较长,胰岛素含量和丙酮酸激酶(PK)活力的提高均相对延迟,而己糖激酶活力的不足又限制了葡萄糖的酵解反应,从而引起血浆胆固醇升高,并伴有持续的血糖和肝糖原升高,从而产生营养胁迫的生理反应,加重鱼类对葡萄糖的代谢负担。  相似文献   

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