植物乳杆菌(Lactobacillus plantarum)对云龙石斑鱼生长、抗氧化性能及血清生化水平的影响

植物乳杆菌(Lactobacillus plantarum)常作为饲料添加剂,可抑制鱼类病原菌生长,增加机体对饲料的消化和吸收。探讨了饲喂不同添加量的植物乳杆菌对云龙石斑鱼(Epinephelus moara♀×E.lanceolatus♂)生长性能、肠道消化酶活性、肠道和肝脏抗氧化能力、血清生化水平的影响,以期为植物乳杆菌在其健康养殖中的应用提供依据。结果显示,随植物乳杆菌的饲喂量增加,云龙石斑鱼的终末均重(FAW)、增重率(WGR)、特定生长率(SGR)和肠道中淀粉酶(AMS)、脂肪酶(LPS)和糜蛋白酶(CHY)、总抗氧化能力(T-AOC)、过氧化氢酶(CAT)、还原型谷胱甘肽(GSH)、超氧化物歧化酶(SOD)的变化均呈现先升高后降低的趋势。而随着植物乳杆菌的饲喂量增加,云龙石斑鱼的血清中总蛋白(TP)、白蛋白(Alb)、胆固醇(TC)、碱性磷酸酶(AKP)、酸性磷酸酶(ACP)、超氧化物歧化酶(SOD)、丙二醛(MDA)均略降低,甘油三酯(TG)略升高,但与对照组差异不明显(P>0.05)。当添加量为1×10⁵CFU/g时,云龙石斑鱼的各生长指标和抗氧化性能均优于对照组,其中增重率显著提高了19.8%(P<0.05),肠道T-AOC、SOD分别显著升高了93.2%、2.9%(P<0.05),肝脏MDA显著降低了32.7%(P<0.05)。当添加量为1×10⁷CFU/g时,云龙石斑鱼的终末均重、增重率、特定生长率均显著高于对照组和其余添加组(P<0.05);饲料系数显著低于对照组(P<0.05);肠道的AMS、LPS、CHY活性分别显著提高了13.0%、166.9%、61.1%(P<0.05);但肠道和肝脏的抗氧化性能变化不明显,仅肝脏MDA显著降低了23.9%(P<0.05)。由此表明,饲料中添加一定量的植物乳杆菌可以提高肠道消化能力和抗氧化能力,促进云龙石斑鱼的生长,但对其血清生化水平无显著性影响,不会对鱼体代谢产生不良作用。     

致病性副溶血弧菌生物膜形成特性研究

采用改良的微孔板法测定了12株弧菌的形成生物膜的效果,选择形成生物膜效果最好的副溶血弧菌ND-02,进一步研究了环境因子对其生物膜形成的影响。实验结果显示副溶血弧菌ND-02菌株在静置培养24～36 h后形成成熟的生物膜,在起始菌浓度为107～108CFU/mL形成生物膜的量最大;在30℃,NaCl浓度为3%～5%,pH偏弱碱性时的生物膜OD590值最大;C 2a+促进副溶血弧菌生物膜的形成,而M 2g+抑制生物膜的形成;副溶血弧菌在分别经大黄鱼表皮黏液、肠黏液和肝脏提取液包被的的基质上形成生物膜的作用明显,鳃黏液和脾脏提取液中次之,肌肉提取液包被后生物膜形成量最低。以上结果表明,副溶血弧菌ND-02菌株能形成稳定而明显的生物膜,而且生物膜的形成受温度、NaCl浓度、pH值等环境因子的影响。     

杜氏鰤的一种细菌病病原菌研究

从患病的杜氏（鱼师）肾脏分离出Y-1菌.人工感染试验,当注射9.0×107CFU/尾时,Y-1菌可致试验鱼100%死亡;注射9.0×106CFU/尾时,死亡率为50%,症状与自然发病相似,结果表明该分离菌为致病菌.Y-1菌为革兰氏阴性,有动力,杆状,极生单鞭毛;发酵葡萄糖,还原硝酸盐,氧化酶、触酶阳性,赖氨酸脱羧酶、鸟氨酸脱羧酶、精氨酸双水解酶阴性.根据形态及生理生化特性,Y-1菌应为弧菌Vibriosp..药敏试验结果表明,该菌对氨基糖甙类、呋喃类、喹诺酮类及大环内脂类药物敏感.     

Immune response of Pseudosciaena crocea to the injection of Vibrio alginolyticus

For the investigation of anti-infection immune response of Pseudosciaena crocea, 160 healthy fish samples were categorized into infected and control groups. Each individual fish in the infected group was injected intraperitoneally (i.p.) with 0.2 ml bacterial suspension of Vibrio alginolyticus in density of 2×107 CFU/ml, while each individual in the control group was injected i.p. with 0.2 ml sterile saline solution (0.85%). It was observed that the artificial injection of V. alginolyticus significantly inc...     

河流弧菌对牙鲆表皮黏液黏附作用的研究

为了解河流弧菌对牙鲆表皮黏液的黏附特性,采用3H-TdR同位素示踪方法研究了细菌浓度、孵育时间、孵育温度、pH值、阳离子、碳水化合物等因子对河流弧菌黏附作用的影响。试验结果表明:河流弧菌能很好地黏附于牙鲆表皮黏液,黏附量随菌浓度升高而增大并符合饱和黏附动力学:y=417.89ln(x)+691.57(R2=0.986);在30℃下黏附量随着孵育时间的延长而增加,180min趋于饱和;孵育温度在4~30℃范围内黏附量随温度升高而增多,30℃时黏附量最大;pH值酸性较碱性黏附作用明显,pH为5时最强;NaCl浓度在0.5%~4.5%范围内,细菌黏附作用随着浓度升高而增强,较纯水存在显著性差异(P<0.05);Ca2+,Mg2+能显著加强河流弧菌的黏附作用,但Ca2+作用明显超过Mg2+;葡萄糖等8种碳水化合物能显著促进河流弧菌的黏附作用(P<0.05)。以上结果说明:河流弧菌对牙鲆表皮黏液的黏附量较大,这有利于进一步感染、致病;河流弧菌的黏附作用受温度、pH值、阳离子、碳水化合物等多种环境因子的影响,较高的水温有利于河流弧菌的黏附,有利于疾病的发生。     

Vaccination in three different ways against vibriosis of <Emphasis Type="Italic">Seriola dumerili</Emphasis> caused by <Emphasis Type="Italic">Vibrio hollisae</Emphasis>

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.     

病原性副溶血弧菌在天然海水中的饥饿耐受研究

将副溶血弧菌培养至对数生长期,用天然海水洗脱,调整菌浓度至107个/cm3左右后置于27℃恒温培养箱进行饥饿试验.结果表明:饥饿初期细菌总数、可培养菌数和活菌数都有较大幅度上升;饥饿中、后期,总菌数与活菌数缓慢下降,而可培养菌数下降速度较快.饥饿前副溶血弧菌呈短杆状,染色均匀;饥饿后许多细胞中央出现染色较浅的区域,细胞形态变为椭圆形.副溶血弧菌对大黄鱼表皮黏液的黏附量随着饥饿时间延长而急剧下降,饥饿7 d后的黏附量接近于空白.用SDS-PAGE分析不同饥饿时期菌体蛋白质的差异,结果表明:饥饿30 d后菌体蛋白条带比饥饿前的蛋白质条带少;饥饿60 d后菌体蛋白质条带比饥饿30 d时的多,但比饥饿前少.研究结果对于揭示副溶血弧菌的流行病学特征有重要的参考价值.     

河流弧菌(Vibrio fluvialis)感染后牙鲆(Paralichthys olivaceus)血清抗菌物质的研究

采用1×107cells/ml的河流弧菌悬液背部肌肉注射感染牙鲆,注射后24h、48h试验组的血清抗菌活力分别极显著性(P0.01)和显著性(P0.05)高于对照组。用Sephadex G-25凝胶柱对第二次人工注射感染后24h牙鲆血清进行分离,其中第7—15收集管对河流弧菌具有抗菌活性,对大肠杆菌、枯草芽孢杆菌等指示菌也具有明显的抗菌作用,在4—100℃范围内,大部分活性分离组分随着温度的升高其抗菌活力有所增强。具有抗河流弧菌活性收集管(第9管)经弱阳离子交换柱分离后,OD280显示出2个蛋白吸收峰,具有抗河流弧菌活性的物质主要集中在第1吸收峰。SDS-PAGE分析显示经Sephadex G-25凝胶柱和弱阳离子交换柱分离的抗菌蛋白分子量较大。以上结果表明,牙鲆被河流弧菌感染后能很快产生大量抗菌物质释放到血清中;主要抗菌物质的分子量较大,且热稳定性较好;这些活性物质的抗菌活性具有较强的特异性。     

拮抗菌对病原性溶藻弧菌(Vibrio alginolyticus)粘附大黄鱼(Pseudosciaena crocea)表皮粘液的影响

采用3H-TdR同位素示踪法研究了病原性溶藻弧菌和9株拮抗菌对大黄鱼表皮粘液的粘附动力学。结果表明,10株细菌的粘附作用都符合饱和粘附动力学特征;R4、R13和R32等3株拮抗菌的最大粘附量高于溶藻弧菌;9株拮抗菌的分离常数都高于溶藻弧菌;R32的亲和指数大于溶藻弧菌,其余8株拮抗菌的亲和指数都小于溶藻弧菌。测定了9株拮抗菌对病原性溶藻弧菌粘附大黄鱼表皮粘液的竞争、排斥和置换作用,结果显示:9株拮抗菌在竞争条件下都能极显著减少溶藻弧菌对表皮粘液粘附(P<0.01);在粘附排斥方面,R4、R13、J26、J28、J312、Y59等6株拮抗菌能显著排斥病原性溶藻弧菌对大黄鱼表皮粘液的粘附作用(P<0.05);在粘附置换方面,J26、J28等2株拮抗菌能极显著(P<0.01)降低病原性溶藻弧菌对大黄鱼表皮粘液的粘附量。研究结果表明,病原性溶藻弧菌对大黄鱼表皮粘液有较强的粘附作用;9株拮抗菌能够抑制溶藻弧菌对大黄鱼表皮粘液的粘附作用,其中竞争作用效果最好,排斥作用次之,置换作用的效果最差;拮抗菌对病原菌粘附的抑制效果受多种因素的影响,各菌株的粘附动力学参数有重要的影响作用。     

Vaccination in three different ways against vibriosis of Seriola dumerili caused by Vibrio hollisae

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.