建立胃癌实验动物模型方法的研究

目的通过对人低分化胃癌细胞系SGC-7901状态和接种细胞数目的研究,建立良好的皮下接种胃癌的动物模型。方法采用腹腔注射SGC-7901细胞,使裸鼠形成腹水;光镜及电镜观察细胞状态;将购买的SGC-7901细胞以及形成腹水后的肿瘤细胞分别以1×108、1×107和1×106个进行裸鼠皮下接种,每组接种5只。观察其肿瘤形成时间、大小、状态及病理学变化。结果SGC-7901细胞接种裸鼠形成腹水后进行培养的肿瘤细胞增殖状态发生改变;购买的SGC-7901细胞以1×108及1×107接种裸鼠,在第21天肿瘤组织中央均出现出血和坏死;1×106的裸鼠第21天未见肉眼可见的肿瘤形成。腹水培养的肿瘤细胞,接种1×108的裸鼠在第21天肿瘤组织中央可见大面积的出血和坏死;接种1×107及1×106的裸鼠在第21天均未见出血和坏死,接种1×107的肿瘤组织体积较大。结论SGC-7901细胞接种裸鼠形成腹水后的细胞,更容易建立SGC-7901细胞皮下接种的胃癌动物模型,其中以接种细胞数为1×107的肿瘤生长较好,更适用于胃癌的实验研究。     

驼乳源益生菌对db/db鼠肠道菌群的调节作用

摘要：目的 探究驼乳来源复合益生菌对db/db鼠血糖及肠道菌群的调节作用。方法 将60只db/db鼠随机分为5组,每组12只,分别为模型组、二甲双胍组、利拉鲁肽组、低剂量复合益生菌组和高剂量复合益生菌组,12只C57BL/Ks小鼠为正常对照组。阳性药组分别灌胃0.3 mg/g二甲双胍及皮下注射0.2 μg/g利拉鲁肽,低剂量复合益生菌组灌胃乳酸菌1.0×108 CFU/d和酵母菌1.0×106 CFU/d,高剂量复合益生菌组灌胃乳酸菌1.0×1010 CFU/d和酵母菌1.0×108 CFU/d,其他组灌胃生理盐水。血糖仪检测空腹血糖（FBG）及不同时间段血糖水平。ELISA法检测糖化血红蛋白（HbA1c）含量。实时荧光定量PCR检测粪便菌群含量。结果 与模型组相比,低、高剂量复合益生菌组FBG水平明显减少,改善葡萄糖耐受（OGTT）能力,降低HbA1c含量。与模型组相比,低、高剂量复合益生菌组显著降低厚壁菌门、放线菌门和大肠埃希菌属水平,增加拟杆菌门、乳杆菌属和双歧杆菌属水平,但对总菌与梭杆菌属无影响。结论 驼乳来源复合益生菌可通过调节肠道菌群降低db/db鼠血糖含量。     

乙型脑炎SA14-14-2减毒株灭活与不灭活以及P3株死疫苗在小鼠体内的免疫原性比较

将乙型脑炎SA14 14 2减毒株 (简称 14 2株 )病毒液和灭活 14 2株病毒液皮下和腹腔免疫小白鼠 ,P3株死苗作对照 ,测免疫效价 ,结果 14 2株病毒液的效力为 1 8× 10 -5～ 6 .1× 10 -6ID50 /ml,灭活 14 2株病毒液为 9 9× 10 -1～ 7 2× 10 -3 ID50 /ml,P3株死苗为 4 0× 10 -3 ～ 2 2× 10 -4 ID50 /ml。同时免后 14天采血用PRNT法测抗体 ,结果为 14 2株病毒液 1∶2 (皮下和腹腔 ) ;灭活 14 2株病毒液 <1∶2 ,P3株死苗为皮下 1∶2、腹腔 1∶10。结果表明 :14 2株在小白鼠体内的免疫原性远远高于灭活 14 2株病毒液和P3株死疫苗 ,14 2株经灭活后对小白鼠基本无保护作用。 14 2株抗体水平不高 ,而免疫力远高于P3株死苗 ,说明 14 2株与P3株死疫苗免疫性质不同 ,有细胞免疫参与。     

尼罗罗非鱼致病性类志贺邻单胞菌(Plesiomonas shigelloides)的分离鉴定及其病理学观察

【目的】2013年5月广东番禺某养殖场罗非鱼出现大量死亡现象,临床症状表现为鱼体体色发黑、体表出血、鳞片脱落、鳍条溃烂等,解剖发现腹腔有大量腹水、胆囊肿大、肝呈黄色、脾脏呈暗红色。为确定病原,【方法】从具以上临床症状的病鱼组织中分离获得可疑菌株1株,编号PYS1。采用形态特征、生长特性、理化特征、16S rRNA基因序列分析等理化及分子生物学技术鉴定该菌株种类。通过人工回归感染及组织病理学研究确定该菌株的致病性,并开展药物敏感性试验筛选其敏感药物。【结果】结果表明PYS1菌株为类志贺邻单胞菌(Plesiomonas shigelloides),在16S rRNA基因序列构建的分子进化树中与其他鱼源类志贺邻单胞菌聚为一支。药敏试验结果表明该菌株已呈现多重耐药性,仅对少数检测抗生素(头孢曲松、头孢洛克和头孢唑啉等)敏感。人工回归感染结果显示PYS1菌株可使尼罗罗非鱼出现与自然发病鱼相似症状,其对尼罗罗非鱼半致死量为1.425×108CFU/尾,石蜡切片显示其对感染鱼的肠、肝、脾、肾和心脏等组织均可造成损伤。【结论】证明类志贺邻单胞菌为上述养殖场尼罗罗非鱼发病的病原,且为首次报道该菌对尼罗罗非鱼的致病性。     

重组人粒细胞集落刺激因子的复性研究

通过小试研究对重组人粒细胞集落刺激因子(rhG-CSF)的复性条件,如氧化剂和还原剂比例、操作方法、时间和蛋白浓度,进行了优化选择,并在此基础上进行了中试放大试验的验证.试验结果表明,采用优化后的复性方法复性液中rhG-CSF的效价可达到1.8×107U/ml以上,比活性达到0.9×108/mg以上.     

实时荧光定量RT-PCR在小鼠及人G蛋白mRNA检测中的应用

用自行设计的TaqMan双标探针和扩增引物建立检测鸟苷酸结合蛋白(G-protein)mRNA的实时荧光定量RT-PCR技术.用G蛋白纯品绘制定量标准曲线,实时荧光定量PCR仪检测ECV304细胞和小鼠G蛋白mRNA水平.10μmol/LGqαmRNA反义寡核苷酸(GqαODN)作用ECV304细胞24h后,Gqα的mRNA表达显著下降(3.18×108±1.75×108拷贝下降到1.44×106±4.82×105拷贝),48h和72h下降更明显;Gsα和Gi2α的mRNA表达变化不显著.10μmol/LGsαmRNA反义寡核苷酸(GsαODN)作用ECV304细胞24h后,Gsα的mRNA表达显著下降(2.97×108±2.68×107拷贝下降到4.16×106±2.00×106拷贝),48h和72h下降更明显;Gqα、Gi2α表达变化不显著.小白鼠油酸致伤后6h,GqαmRNA表达显著下降(1.16×108±8.73×106拷贝下降到3.30×106±1.68×106拷贝),24h下降更显著(9.32×107±1.47×107拷贝下降到4.14×106±1.67×106拷贝);Gsα和Gi2α表达变化的趋势同GqαmRNA.结果准确可靠,重现性好.说明建立的实时荧光定量RT-PCR方法成功地实现了对ECV304细胞和小白鼠肺组织G蛋白不同亚型不同丰度基因表达的检测.     

柠檬形克勒克酵母对温州蜜柑“国庆一号”采后贮藏的防腐效果

Citrus unshiu “Guoqing No.1” was used as material for elucidating the effect of Kloeckera apiculata suspension post-harvest treatment on preventing fruit decay during storage period. The result showed that K. apiculata preparation of 2×106CFU/mL (CFU, cloning forming unit) significantly reduced the development of decay caused by green mold, blue mold and stem rot, showing the same effect of carbendazim treatment after storage period of 100 days, and the disease incidence was controlled under 1%. The antagonist population was between 3.5×105CFU/mL and 1.7×107CFU/mL in citrus fruit, and increased 1.7×107CFU/mL after 40 days storage, and then stabilized for the remaining storage period. After storage at 5℃ for 100 days, K. apiculata strain 34-9 did not alter any quality parameters of fruit. During storage, the change of relative electrical conductivity (REC) showed a coordinate tendency with malondlaldehyde (MDA) content, which kept continuously growing from the beginning to the end of the storage. In the storage period of 25 days, the REC value and MDA content of the treatments were all significantly lower than those of water treatment, but had no obvious difference from those of carbendazim treatment. K. apiculata strain 34-9 had no effect on the enzyme activity of super oxide dismutase (SOD), but significantly improved peroxidase (POD) activity after 100 day storage.     

海南罗非鱼无乳链球菌分离鉴定及其特性研究

从海南省患暴发性疾病的罗非鱼(Tilapia)上分离出1株细菌HNLFYL4,对分离菌株进行了鉴定及致病性和药物敏感性研究.通过形态学观察和生理生化鉴定,结果显示,分离菌株为无乳链球菌(Streptococcus agalactiae).对分离菌株的16S rRNA基因进行PCR扩增和测序,所得序列已登录到GenBank,登录号为HQ645983,与GenBank中收录的链球菌16S rRNA 基因进行比对并构建系统进化树,结果表明,分离菌株的16S rRNA基因序列与无乳链球菌同源性高达100%,进一步确定分离菌株为无乳链球菌.人工感染显示分离菌株对小白鼠和罗非鱼均具有致病性,对小白鼠的LD50为1.0×104 CFU/mL,对体重为500g±20g的罗非鱼的LD50为1.729×109CFU/mL.分离菌株对氯霉素、青霉素G、呋喃妥因等敏感,对丁胺卡那、链霉素、卡那霉素等不敏感.     

眼斑拟石首鱼重复DNA序列的染色体定位

针对眼斑拟石首鱼Sciaenops ocellatus染色体标记匮乏的问题, 利用荧光原位杂交(FISH)定位了眼斑拟石首鱼的18S rDNA、5S rDNA和端粒序列。结果显示, 眼斑拟石首鱼的核型公式为2n=48t; 仅有1对18S rDNA位点, 位于第1对染色体的次缢痕部位; 有2对5S rDNA位点, FISH信号强度不等, 强信号位于第8对染色体的近着丝粒端, 弱信号位于第3对染色体的臂间。端粒FISH信号出现于所有染色体的两端, 但表现出染色体两端信号不平衡的特点, 着丝粒端FISH信号明显强于远端信号。这一特点为判定染色体的方向提供了便利。结合其他石首鱼的核型数据可以推断, 2n=48t的核型及单对近着丝粒分布的18S rDNA位点是石首鱼的共同祖征; 在石首鱼进化过程中, 曾发生活跃但不影响宏观核型的小规模重排。研究结果丰富了眼斑拟石首鱼染色体的辨识标记, 并为研究石首鱼染色体进化提供了基础数据。     

Gateway技术构建交链孢菌JH505 cDNA文库

Gateway(R)技术构建Cdna文库,利用λ噬菌体的位点特异性重组,避免使用限制性内酶切割Cdna,能够解决常规方法构建Cdna文库的技术缺陷.首次应用Gateway(R)技术构建交链孢菌Cdna文库,经检测Cdna入门文库的滴度达到1×107cfu/Ml,文库总容量为9×107cfu,平均插入片段为1510bp.通过LR重组把入门文库转换为表达文库,表达文库的滴度为1.58×106cfu/Ml,文库总容量为6.32×106cfu,平均插入片段大小为1680bp.表达文库的构建为进一步克隆植物激活蛋白基因打下了基础.     

Red drum Sciaenops ocellatus mortalities associated with Streptococcus iniae infection.

We isolated for the first time Streptococcus iniae strains associated with diseased marine fish. Diseased red drum Sciaenops ocellatus were lethargic, and presented external signs (exophthalmia and loss of orientation) resembling those of freshwater fish infected by S. iniae. Skin lesions, extending to a necrotizing myositis, were typical of S. iniae infection of red drum. Histopathological findings indicate that S. iniae infection in red drum produces a chronic disease with systemic involvement characterized by multiple necrotic foci. Molecular epidemiology (RFLP [restriction fragment length polymorphism] ribotyping) revealed that 2 different ribotypes were involved in a single outbreak. The first is the EcoRI 'Israeli' trout and tilapine ribotype (Hind III type a strains), while the second is the EcoRI 'American' ribotype (Hind III type b strains), typical of tilapines farmed in Texas and Idaho.     

Streptococcus iniae infections in Red Sea cage-cultured and wild fishes

Streptococcus iniae was isolated from 2 moribund wild Red Sea fishes, Pomadasys stridens (Pomadasyidae) and Synodus variegatus (Synodontidae), both collected in shallow waters along the Israeli coast of the Gulf of Eilat. The site is approximately 2 km from a mariculture cage farm in which streptococcal infections were diagnosed in previous years in the red drum Sciaenops ocellatus. This is the first report of S. iniae in Red Sea fishes. Biochemical and molecular similarities between the isolates from cultured fishes and those from the wild specimens suggest that a single strain is involved, and that 'amplification' and dispersal of this pathogen from captive to feral fishes have occurred. At the molecular level, the pathogen is different from the S. iniae isolates that have been afflicting the Israeli freshwater aquaculture in recent years. Although S. iniae prevalence in the wild fish populations of the area remains to be determined, the northernmost region of the Gulf of Eilat, virtually landlocked and with generally calm seas and weak currents, seems to be particularly vulnerable to the impact of diseases that develop in this mariculture system.     

环丙沙星预先处理牙鲆消化道对干酪乳杆菌定植的影响

利用1株干酪乳杆菌,通过实验研究用环丙沙星预先处理牙鲆消化道后乳杆菌的定植和演替规律。在投喂含有1.2×10^9CFU/g乳杆菌的饲料5 d后,消化道定植的乳杆菌超过106CFU/g,其后维持在10^6～10^8CFU/g动态平衡中。同时随着乳杆菌的投喂,不经环丙沙星预先处理牙鲆消化道的正常组,鱼消化道的弧菌数从10^7～8CFU/g降低到10^6CFU/g左右;而经环丙沙星预先处理牙鲆消化道的药饵组,鱼胃、小肠和盲囊的弧菌数则是先增加,然后显著下降。停喂乳杆菌7 d后,2个实验组鱼消化道的乳杆菌均从10^5～6CFU/g下降到10^4CFU/g,干酪乳杆菌正常组鱼盲囊中弧菌从10^5CFU/g回升到10^6CFU/g,胃和小肠中弧菌数量基本不变。干酪乳杆菌药饵组则有所不同：除胃中弧菌数量则基本不变外,盲囊和小肠中弧菌数量继续下降,其原因有待进一步研究。实验结果表明,干酪乳杆菌能在牙鲆消化道内定植,而用预先处理牙鲆消化道后,更有利于乳杆菌的定植;乳杆菌的投喂和定植,使牙鲆消化道中的弧菌数量明显下降。     

电解海水的抑菌活性及对食品加工表面材料的消毒效果

为了考察直接电解海水消除细菌污染的可能性,本文将海水及海水稀释成不同浓度后通过氧化电解水装置进行电解不同时间后,所得酸性电解海水、碱性电解海水和中性电解海水对病原菌[埃希氏大肠杆菌(Escherichina coli)、沙门氏菌(Salmonella)、单核细胞增生李斯特菌(Listeria moncytogene)、摩化摩根(Morganella morganii)、副溶血性弧菌(Vibrio parahaemolyticus)]以及食品加工表面接触材料(地板砖、不锈钢板、瓷砖、手套、抹布)的消毒效果进行分析研究.结果表明,酸性电解海水具有良好的杀菌效果,能将107 CFU/mL的病原菌悬液在1 min内几乎全部杀死.碱性电解海水和中性水无明显的杀菌效果.通过模拟食品加工过程,对食品加工表面接触材料人为染菌,研究电解海水对表面材料的消毒效果,结果表明酸性电解海水仍能将表面材料含有的107CFU/cm2病原菌在5 min之内几乎全部杀灭.由此说明电解海水对食品加工表面接触材料具有明显的消毒效果,能取代以淡水为原料的电解水杀菌效果是高效廉价和不浪费淡水资源的一种理想消毒剂.     

Chitosan nanogel aqueous treatment improved blood biochemicals,antioxidant capacity,immune response,immune-related gene expression and infection resistance of Nile tilapia

Candida albicans is a pathogenic yeast recently associated with diverse diseases in aquaculture. The present study investigated the efficacy of chitosan nanogel (CNG) in ameliorating effects of C. albicans on Nile tilapia (Oreochromis niloticus). Fish were randomly distributed into four groups (control, waterborne CNG at 75 μg/L, intraperitoneally challenged with C. albicans (1.8 × 107 CFU/mL), and waterborne CNG at 75 μg/L + C. albicans at 1.8 × 107 CFU/mL). Results showed that C. albicans infection reduced survival rate (57.5%) and caused marked clinical symptoms in fish among all infected groups. Substantial declines in immunological (complement 3, lysozyme, and immunoglobulin M), protein (total protein and non-albumin protein), and antioxidant (catalase, glutathione peroxidase, and superoxide dismutase) biochemical endpoints were exhibited, The C. albicans infected group also showed marked down-regulation in the expression of immune-related genes, including toll-like receptor 2 (TLR-2), transforming growth factor beta2, tumor necrosis factor alpha, interleukins (IL-1β, IL-6, and IL-10), and antiapoptotic gene (B-cell lymphoma, BCL-2). The expression of the apoptotic gene (Bcl-2 associated X protein, BAX) was up-regulated in fish challenged by C. albicans. The application of waterborne CNG to fish challenged with C. albicans infection improved fish survival (79.5%) and all other measured parameters. The main finding of this work was that CNG is a nanotechnology with potential for preventing degraded health status by C. albicans infection in tilapia, and thus has promise as an intervention in aquaculture settings.     

Infectious gastroenteritis caused by Vibrio harveyi (V. carchariae) in cultured red drum, Sciaenops ocellatus

Summary An outbreak of serious mortality among the cultured red drum Sciaenops ocellatus (L.) characterized by a swollen intestine containing transparent yellow fluid (ascites and gastroenteritis) occurred in July 2000 in Taiwan. A motile strain Rd 0700 was isolated from head kidney and/or the intestinal yellow fluid on tryptone soya agar (TSA) supplemented with 2% (w/v) NaCl and/or thiosulfate citrate bile salt (TCBS) sucrose agar plates. Applying biochemical characteristics, this strain was characterized and identified as Vibrio harveyi (V. carchariae). The bacteria could be re‐isolated from kidney, liver, and the transparent yellow fluid of swollen intestine of fish after bacterial challenge. The LD₅₀ values of the organism and its extracellular products (ECP) were 2.9×10⁷ colony forming units (CFU) and 3.85 μg protein g^?1 fish body weight, respectively. All moribund/dead fish exhibited gastroenteritis except those killed within 12 h. This is a first report showing that intraperitoneal (i.p.) injection of the ECP from V. carchariae is lethal to red drum and can reproduce gastroenteritis in the fish.     

In vivo activation of tilapia nonspecific cytotoxic cells by Streptococcus iniae and amplification with apoptosis regulatory factor(s)

An important component of immediate innate responses of tilapia to stress is the release within minutes of soluble cytokine-like substances into the peripheral circulation. These cytokine-like stress factors bind nonspecific cytotoxic cells (NCC) and produce 3-4-fold increased cytotoxicity. In the present study, the in vivo responses of tilapia NCC following injection with different isolates of intact killed Streptococcus iniae was investigated. Activated cytotoxicity of NCC in the peripheral blood (PB) was produced by increased specific activity of resident cells rather than increased numbers. Tilapia injected intravenously (i.v.) with killed S. iniae produced different cytotoxicity responses compared to fish injected intraperitoneally (i.p.). In the spleen (S) and anterior kidney (AK), there was no correlation between S. iniae isolate and cytotoxicity response at 4, 8 or 24 h following i.p. injection. The NCC response following i.v. injection of killed bacteria was different. Within minutes following i.v. injection, NCC cytotoxicity from the PB increased 100% compared to naive controls. The existence of subsets of differentiated NCC in the PB was suggested because i.v. injection had no amplification effects on NCC from the AK or S. Likewise, NCC from the PB only appeared to exhibit a degree of antigen specificity. S. iniae strain #173 produced activation of cytotoxicity compared to isolates #164 and ATCC. Evidence for soluble factor (cytokine?) involvement in increased cytotoxicity was obtained by passive activation of NCC with serum from #173 (i.v.) injected fish. Incubation of this serum with control (na?ve) NCC produced large increases in the cytotoxicity of labelled HL-60 target cells. Similarly obtained serum from fish injected with ATCC and #164 isolates had no amplification activity. Studies were also performed to study the mechanism(s) of passive activation. Flow cytometric analysis revealed that NCC from the S, AK and PB constitutively expressed cytosolic (not membrane) FasL. Stress serum treated NCC obtained from the peripheral blood produced an increase in the expression of FasL, CAS and FADD by Western blot examination. These data indicated that cytokine like factors in the serum of stressed tilapia activate increased NCC cytotoxicity (possibly) by stimulating the expression of proteins involved in activation of programmed cell death.     

Streptococcus iniae inhibition of apoptosis of nonspecific cytotoxic cells: a mechanism of activation of innate immunity in teleosts

Nonspecific cytotoxic cells (NCC) may provide innate anti-bacterial resistance against Streptococcus iniae infections in tilapia. The mechanism of immunity would be elaboration and release of various cytokines, augmentation of inflammation and amplification of increased antigen processing. To investigate bacterial regulation of NCC function, 2 different processes of cellular pathology were examined: apoptosis and necrosis. Different isolates of S. iniae from diseased teleosts, a dolphin and a human were tested. All isolates were examined for their ability to produce apoptosis and/or necrosis on freshly purified tilapia NCC and on a tilapia continuous cell line (i.e. TMB-8 cells). Two different isolates (9033 and 173) inhibited the outer membrane expression of phosphatidylserine (PS) by NCC, an early sign of apoptosis. This occurred at 4 h post-treatment and lasted throughout the 24 h treatment period. All other isolates either did not differ from control levels or produced a small increase in PS expression by NCC. The early reduction in PS expression occurred concomitantly with increased necrosis associated with nonspecific DNA fragmentation. Two-color flow cytometry (Annexin-V vs propidium iodide staining) demonstrated the specificity of Annexin-V binding. Experiments were also done to determine the effects of S. iniae on TMB-8 cells. Treated TMB-8 cells did not produce appreciable Annexin-V binding. Compared to the ATCC strain, 9033 produced high levels of necrosis-associated DNA fragmentation of TMB-8 cells at 4 and 8 h post-treatment. These data indicated that different isolates of S. iniae may regulate NCC anti-bacterial resistance by causing reduced levels of programmed cell death (PCD), increased necrosis and associated enhancement of inflammatory responses. Understanding the relevance of these bacterial effects on NCC may be an important consideration in the evaluation of isolates used in vaccine/ bacterin production.     

玉米青贮过程中乳酸菌动态变化

为了揭示青贮玉米在发酵过程中乳酸菌数量及种群的动态变化,以及微生物添加剂对乳酸菌种群变化的影响,采用了培养方法计数发酵过程中乳酸菌数目的变化,利用16S rRNA基因序列比对方法分析青贮玉米中乳酸菌的多样性及种群变化趋势。经过对15d时间内青贮玉米中乳酸菌变化趋势的分析显示:一周后对照组乳酸菌数最高达到2.1×106CFU/g,两处理组中处理组Ⅱ乳酸菌数达到最高5.5×107CFU/g;利用MRS平板分离、培养出典型乳酸菌菌落152株,经16S rRNA基因序列比对分析为乳杆菌属和片球菌属,其中86%的菌株属于乳杆菌属。此研究表明微生物添加剂有利于青贮玉米发酵过程中乳酸菌的快速增殖,乳杆菌属和片球菌属都是青贮玉米发酵的启动菌之一,在发酵前期一直存在,但发酵后期乳杆菌属是玉米青贮过程中乳酸菌的主要菌群。     

Inflammatory responses of Nile tilapia Oreochromis niloticus to Streptococcus agalactiae: effects of vaccination and yeast diet supplement

This study evaluated the effects of dietary supplementation with 0.3% Saccharomyces cerevisiae yeast cell wall and of vaccination against Streptococcus agalactiae on the cellular component of acute inflammation induced in the coelomic cavity of Nile tilapia Oreochromis niloticus and on survival of the fish after challenge. A total of 84 tilapia of mean (±SD) weight 125.0 ± 1.5 g were distributed among twelve 310 l fiberglass tanks according to a 2 × 2 × 3 factorial design in the following manner: with and without supplementation; 2 stimulations (oily solution without S. agalactiae vaccine and vaccination); 15 d later all fish were intracoelomically challenged with 108 CFU ml-1 of a homologous strain of S. agalactiae, and evaluated after 6, 24 and 48 h, with 7 replicates. The fish received the non-supplemented or supplemented diet for a total of 77 d. The vaccination was performed on the 60th day, intracoelomically, as a single injection of 0.5 ml of the vaccine containing 108 CFU ml-1. Fifteen days later, all the fish were challenged with S. agalactiae by means of an intracoelomic inoculation of 108 CFU ml-1. No mortality was observed among the supplemented fish. The fish that were fed the non-supplemented diet and immunized with the bacterium presented a mortality rate of 28.5%. Among the non-supplemented and non-immunized fish, the mortality rate was 38.09%. Supplementation, in both vaccinated and non-vaccinated fish, induced larger accumulations of thrombocytes, lymphocytes and macrophages at the inflammatory focus. The results suggest that supplementation with 0.3% yeast cell wall, in both vaccinated and non-vaccinated fish, improved the inflammatory response of the fish and protected against the challenge. Vaccination increased the defense response, but the effect was stronger when associated with supplementation with S. cerevisiae.