Thompson D R, Parreira V R, Kulkarni R R, Prescott J F
Department of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph, Ont., Canada N1G 2W1.
Vet Microbiol. 2006 Mar 10;113(1-2):25-34. doi: 10.1016/j.vetmic.2005.10.015. Epub 2005 Nov 11.
A vaccine for necrotic enteritis (NE) of chickens would reduce the current need to prevent or treat the disease in broiler chickens with antimicrobial drugs. The objective of this study was to understand aspects of immunity to the disease. The first experiment examined the virulence of six strains of Clostridium perfringens isolated from cases of NE in broiler chickens. Using a 5-day experimental oral infection of 2-week-old broiler chickens, four of the six strains were found to be virulent. Pulsed-field gel electrophoresis and PCR showed that virulence was not associated with a plasmid encoding the beta2 toxin gene, cpb2, since this was present in virulent and one of the two avirulent strains. In the second experiment, two virulent and one avirulent strains were tested for their ability to immunize ("infection-immunization") chickens through the oral route. The procedure used experimental infection for 5 days followed by bacitracin treatment for 9 days, and then re-challenge 2 days later with a virulent strain, CP4. Infection-immunization with the virulent isolates protected chickens from subsequent virulent challenge, whereas the infection-immunization with the avirulent isolate did not. In a third experiment, two of four alpha-toxin-negative mutants of CP4 protected birds from experimental NE after oral immunization. These two mutants were also attenuated for virulence. We conclude that it is possible to immunize chickens successfully against NE and that immunogen(s) other than alpha-toxin are important in protective immunity against oral infection.
一种针对鸡坏死性肠炎(NE)的疫苗将减少目前使用抗菌药物预防或治疗肉鸡该疾病的需求。本研究的目的是了解针对该疾病的免疫方面。第一个实验检测了从患有NE的肉鸡病例中分离出的6株产气荚膜梭菌的毒力。通过对2周龄肉鸡进行为期5天的实验性口服感染,发现6株中的4株具有毒力。脉冲场凝胶电泳和聚合酶链反应表明,毒力与编码β2毒素基因cpb2的质粒无关,因为该质粒存在于有毒力的菌株以及两株无毒力菌株中的一株中。在第二个实验中,测试了两株有毒力菌株和一株无毒力菌株通过口服途径对鸡进行免疫(“感染-免疫”)的能力。该程序采用5天的实验性感染,随后用杆菌肽治疗9天,然后在2天后用有毒力的CP4菌株再次攻击。用有毒力的分离株进行感染-免疫可保护鸡免受随后的有毒力攻击,而用无毒力的分离株进行感染-免疫则不能。在第三个实验中,CP4的四个α毒素阴性突变体中的两个在口服免疫后保护鸡免受实验性NE的侵害。这两个突变体的毒力也有所减弱。我们得出结论,成功免疫鸡抵抗NE是可能的,并且除α毒素外的免疫原在针对口服感染的保护性免疫中很重要。