作　　者： (霍建强）; (赵玲敏）; (覃映雪）; (鄢庆枇）;

机构地区： 集美大学水产学院,福建厦门361021 农业部东海海水健康养殖重点实验室,福建厦门361021

出　　处： 《集美大学学报（自然科学版）》 2017年第4期12-19,共8页

摘　　要： 选取甲醛、苯酚、和氯仿三种化学灭活剂在不同的条件下对变形假单胞菌进行灭活处理,确定了0.2%甲醛溶液28℃处理12 h;0.5%苯酚溶液28℃处理24 h;3.5%氯仿溶液28℃处理12 h的灭活条件。分别用三种灭活剂制备的疫苗免疫斜带石斑,测定斜带石斑的血清抗体效价和感染后的免疫保护率。结果显示:接种甲醛灭活疫苗组的血清抗体效价最高,接种苯酚灭活疫苗组的抗体效价次之,接种氯仿灭活疫苗组的抗体效价最低;三种灭活疫苗的免疫保护率分别为甲醛灭活疫苗46.67%,苯酚灭活疫苗30%,氯仿灭活疫苗20%。选择甲醛灭活疫苗与弗氏佐剂、白油佐剂、氢氧化铝、206佐剂、蜂胶等5种佐剂分别配伍后注射免疫斜带石斑,结果显示:添加佐剂组斜带石斑血清抗体效价全都大幅高于未添加佐剂组,其中白油的效果最好;添加佐剂组免疫保护率也全都大幅高于未添加佐剂组,也是白油佐剂的效果最好,免疫保护率高达91.67%,其余各组分别为氢氧化铝83.33%、206佐剂80.56%、弗氏佐剂72.22%、蜂胶66.67%。研究表明研制出了具有良好免疫保护效果的变形假单胞菌灭活疫苗,为进一步的中试试验奠定了基础。 In the present study, three chemical inactivation agents （methanol, phenol and chloroform） were used to investigate the inactivation condition of Pseudomonas plecoglossicida, and the selected conditions were as follows： 0.2% methanol for 12 h at 28 ℃, 0. 5% phenol for 24 h at 28 ℃, and 3.5% chloroform for 12 h at 28 ℃. In order to estimate the application of the inactivated vaccine in aquaculture, a trial was designed to investigate the effect of inactivated vaccine of P. plecoglossicida on serum antibody titer and im- mune protection after infection in grouper Epinephelus coioides. The highest antibody titer was observed in fish injected with formalin-killed vaccine, while the lowest was obtained in fish injected by chloroform-killed vac- cine. The immune protection rate of the three inactivated vaccine were 46.7% of formalin - killed vaccine. 30% of phenol-killed vaccine and 20% of chloroform - killed vaccine, respectively. In order to investigate the effects of different immune adjuvants, fish were injected by formalin - killed vaccine and five different adjuvants respectively, including Freund＇s adjuvant, white oil adjuvant, alhydrogel, 206 adjuvant and propolis adjuvant. The results showed that both of the serum antibody titer and immune protection rate of E. coioides injected by adjuvant added groups were higher than the control group without adjuvant, and the white oil adju- vant treated group exhibited the highest antibody titer and immune protection rate among the treatment groups. The results indicated that the inactivated vaccine of P. plecoglossicida could effectively improve the immune response of E. coioides. The study provides a theoretical basis for further pilot test.

关 键 词： 变形假单胞菌 灭活疫苗 斜带石斑 免疫佐剂