Shafiq Muhammad Hassan, Kamble Nitin M, Kim Tae Hoon, Choi Yoonyoung, Lee John Hwa
College of Veterinary Medicine and Bio-Safety Research Institute, Chonbuk National University, Iksan, 570-752 Korea.
Avian Dis. 2015 Dec;59(4):537-42. doi: 10.1637/11243-072815-Reg.1.
Live attenuated vaccines are used for effective protection against fowl typhoid (FT) in domestic poultry. In this study, a lon/cpxR/asd deletion mutant of Salmonella Gallinarum expressing the B subunit of a heat labile toxin (LTB) from Escherichia coli, a known adjuvant, was cloned in a recombinant p15A ori plasmid, JOL1355, and evaluated as a vaccine candidate in chickens. The plasmid was shown to be stable inside the attenuated Salmonella Gallinarum cell after three successive generations. Moreover, from an environmental safety point of view, apart from day 1 the JOL1355 strain was not detected in feces through day 21 postinoculation. For the efficacy of JOL1355, a total of 100 chickens were equally divided into two groups. Group A (control) chickens were intramuscularly inoculated with phosphate-buffered saline at 4 and 8 wk of age. Group B chickens were primed and boosted via the intramuscular route with 200 μL of a bacterial suspension of JOL1355 containing 1 × 10(8) colony forming units. All the chickens in Group A and B were challenged at 3 wk postbooster by oral inoculation with a wild-type Salmonella Gallinarum strain, JOL420. The JOL1355-immunized group showed significant protection and survival against the virulent challenge compared to the nonimmunized group. In addition, Group B exhibited a significantly higher humoral immune response, and the chickens remained healthy without any symptoms of anorexia, diarrhea, or depression. Group B also exhibited a significantly lower mortality rate of 4% compared to the 46% of the control group, which can be attributed to higher immunogenicity and better protection. The Group B chickens had significantly lower lesion scores for affected organs, such as the liver and spleen, compared to those of the control chickens (P < 0.01). These findings suggest that JOL1355 is a promising candidate for a safe and highly immunogenic vaccine against FT.
减毒活疫苗用于有效预防家禽的禽伤寒(FT)。在本研究中,将表达来自大肠杆菌的一种已知佐剂——热不稳定毒素(LTB)B亚基的鸡沙门氏菌lon/cpxR/asd缺失突变体克隆到重组p15A ori质粒JOL1355中,并在鸡中作为候选疫苗进行评估。该质粒在连续传代三代后在减毒鸡沙门氏菌细胞内显示稳定。此外,从环境安全性角度来看,除接种后第1天外,在接种后第21天的粪便中均未检测到JOL1355菌株。对于JOL1355的效力,将总共100只鸡平均分为两组。A组(对照组)鸡在4周龄和8周龄时肌肉注射磷酸盐缓冲盐水。B组鸡通过肌肉途径用200μL含1×10⁸菌落形成单位的JOL1355细菌悬液进行初免和加强免疫。A组和B组的所有鸡在加强免疫后3周通过口服接种野生型鸡沙门氏菌菌株JOL420进行攻毒。与未免疫组相比,JOL1355免疫组在抵抗强毒攻击方面显示出显著的保护作用和存活率。此外,B组表现出显著更高的体液免疫反应,并且鸡保持健康,没有任何厌食、腹泻或抑郁症状。B组的死亡率也显著低于对照组,为4%,而对照组为46%,这可归因于更高的免疫原性和更好的保护作用。与对照鸡相比,B组鸡受影响器官如肝脏和脾脏的病变评分显著更低(P<0.01)。这些发现表明JOL1355是一种有前途的针对禽伤寒的安全且高免疫原性疫苗候选物。