Detection and identification of adhesins involved in adhesion of Campylobacter jejuni to chicken skin.

Campylobacter jejuni is the leading cause of bacterial food poisoning worldwide. Chickens are considered to be one of the major reservoirs of Campylobacter infection in humans due to colonization of their intestinal tract. When the chickens are slaughtered and processed, the entire skin of the carcass becomes contaminated with campylobacters. We observed that the number of C. jejuni attached to chicken skin was reduced significantly after treatment of the skin with sodium hydroxide followed by washing with PBS, implying that adhesion factors involved in binding to C. jejuni may exist on skin. Such potential binding-related proteins present in alkaline extracts of the skin surface were detected by a two-dimensional overlay assay and identified by liquid chromatography mass spectrometry (LC-MS). Chicken serum albumin (CSA) was identified as a major protein in these alkaline extracts and confirmed by ELISA to bind specifically to C. jejuni. Moreover, using the same approach, flagellar hook protein E (FlgE) and major outer membrane protein (MOMP) in C. jejuni were identified as bacterial adhesins that bound to the CSA. The ability to bind CSA was also confirmed using recombinant FlgE and MOMP of C. jejuni expressed in Escherichia coli. The present findings suggest that adhesins expressed on C. jejuni cells may bind specifically via proteins present on the skin, as well as by physical attachment.

Aerotolerance and multilocus sequence typing among Campylobacter jejuni strains isolated from humans, broiler chickens, and cattle in Miyazaki Prefecture, Japan.

Campylobacter jejuni is one of the leading causes of human gastroenteritis in Japan. As chickens and cattle are common reservoirs for C. jejuni, this microaerophilic, stress-sensitive bacterium can overcome and survive various stress conditions during zoonotic transmission, particularly foodborne, to humans. How C. jejuni overcomes stress conditions is, however, unclear. In the present study, 70 C. jejuni strains isolated from various sources (26 human, 20 broilers, and 24 cattle isolates) in Miyazaki, Japan, from 2010 to 2012, were subjected to multilocus sequence typing (MLST) and aerotolerance testing (aerobic shaking at 200 rpm). The results demonstrated that C. jejuni strains from Miyazaki belonged to 12 clonal complexes (CCs) and 43 sequence types (STs). CC-21 and CC-460 were mainly detected in human clinical strains. Most tested strains were aerotolerant, and only one (1.4%) was deemed sensitive to aerobic stress. Approximately 40% strains survived the 24-hr vigorous aerobic shaking at 200 rpm, and these hyper-aerotolerant strains were more prevalent in broiler and cattle isolates than in human isolates. Phylogenetic analysis divided the strains into five clusters, each showing a different pattern of host association. Thus, we have demonstrated for the first time that C. jejuni strains with increased tolerance to aerobic stress are highly prevalent in broilers and cattle in Miyazaki, Japan, and that certain clonal populations are frequently implicated in human infection in this area.