A fundamental study of the contamination of C. jejuni at a broiler farm and the detection of C. jejuni by means of polymerase chain reaction method

ABSTRACT Campylobacter jejuni (C. jejuni) is a major causative agent of human diarrheal disease. In this study, the survey of the dissemination of C. jejuni in a broiler farm and the infection route of C. jejuni to the broiler was carried out. C. Jejuni was isolated from 67 fecal matters of 101 specimens in one broiler farm. The survey of the transmission route of C. jejuni was carried out on three flocks in this farm. A flock was kept in the windowless house, and the workers changed their clothes and shoes when they went into the poultry house. B and C flocks were kept in the usual house, and the workers changed only their shoes. C. jejuni wasn't isolated in the one week old chickens, but it was isolated from all of the samples from 3 weeks old chicken in B and C flocks, and was widely detected at each week of age throughout the servey period in B and C flocks. In A flock, C. jejuni was isolated from only two chickens of four weeks of age and one chicken of seven weeks of age. These serotypes were mostly J-5. C. jejuni isolation happened after the shipping H flock which was contaminated by J-5. When the shipping, the workers did not change clothes or shoes. Therefore, it was considered that C. jejuni was transmitted through workers. Polymerase chain reaction method (PCR) was applied for the detection of C. jejuni, and specific detection of C. jejuni was achieved By PCR with a pair of primers from the flagellant a gene (flap A). The detection limit in chicken litter was 10E2.0 CFU/g. PCR products of C, jejuni used for the serotyping were digested with MboI. 4 patterns were obtained, type-1 and 2 products were considered the fla A gene reported previously, and suggested the ability to divide C. jejuni into at least four types by restriction fragment length polymorphism (RFLP). The correlation between the results Lior serotyping and PCR-RFLP was carried out in Lior 4, Lior 6, TCK 12, and Lior 7. And concluded that the PCR-RFLP results of this study and the sero-typing results were quite the same. PCR-RFLP with this primer pair was considered to have potential for epidemiological surveys and the detection of the source of food poisoning due to C. jejuni.