Potential use of characterised hyper-colonising strain(s) of Campylobacter jejuni to reduce circulation of environmental strains in commercial poultry.

Sixty-two persistently colonising Campylobacter jejuni strains were tested for their ability to dominate colonisation of the chicken gastrointestinal tract in competition with each other leading to selection of dominant or "hyper-colonising"Campylobacter strains, which are able to displace others in the chicken intestinal tract. One such strain was shown to be a hyper-efficient coloniser of chickens, as it was able to displace other colonising strains, as well as maintain itself in the chicken intestinal tract for the duration of the 56-day broiler production cycle. Once colonisation was established, this hyper-colonising C. jejuni strain, 331, could not be displaced by other colonising or hyper-colonising strains. We proposed that a defined, hyper-colonising strain, or a cocktail of defined strains with a similar phenotype, could form the basis for biological control of unknown/uncharacterised Campylobacter strains from the environment that continuously colonise chicken flocks. To validate this approach, three different chicken infection trials were carried out. These trials demonstrated that the dominant strain of C. jejuni was able to colonise broiler chickens consistently and for the entire life of the birds irrespective of the day of inoculation and antimicrobial agents used in the feed to control other pathogenic micro-organisms. In addition, we have shown that the bio-control strain was able to replace other colonising strains at various points of a 56-day broiler production cycle irrespective of time and type of inoculation. This strain was also capable of re-establishing itself following the challenge with other strains, with and without re-challenge. This work represents a "proof of principle" that a defined C. jejuni strain could be used to biologically control circulation of uncharacterised environmental strains in commercial poultry flocks.

Campylobacter jejuni biofilms up-regulated in the absence of the stringent response utilize a calcofluor white-reactive polysaccharide.

The enteric pathogen Campylobacter jejuni is a highly prevalent yet fastidious bacterium. Biofilms and surface polysaccharides participate in stress survival, transmission, and virulence in C. jejuni; thus, the identification and characterization of novel genes involved in each process have important implications for pathogenesis. We found that C. jejuni reacts with calcofluor white (CFW), indicating the presence of surface polysaccharides harboring beta1-3 and/or beta1-4 linkages. CFW reactivity increased with extended growth, under 42 degrees C anaerobic conditions, and in a DeltaspoT mutant defective for the stringent response (SR). Conversely, two newly isolated dim mutants exhibited diminished CFW reactivity as well as growth and serum sensitivity differences from the wild type. Genetic, biochemical, and nuclear magnetic resonance analyses suggested that differences in CFW reactivity between wild-type and DeltaspoT and dim mutant strains were independent of well-characterized lipooligosaccharides, capsular polysaccharides, and N-linked polysaccharides. Targeted deletion of carB downstream of the dim13 mutation also resulted in CFW hyporeactivity, implicating a possible role for carbamoylphosphate synthase in the biosynthesis of this polysaccharide. Correlations between biofilm formation and production of the CFW-reactive polymer were demonstrated by crystal violet staining, scanning electron microscopy, and confocal microscopy, with the C. jejuni DeltaspoT mutant being the first SR mutant in any bacterial species identified as up-regulating biofilms. Together, these results provide new insight into genes and processes important for biofilm formation and polysaccharide production in C. jejuni.

Comparison of 2-day-old and 14-day-old chicken colonization models for Campylobacter jejuni.

In this study, we compared two types of chicken infection models for Campylobacter jejuni in terms of infectious dose required to colonize the chickens and the susceptibility of chickens of different ages to persistent colonization by C. jejuni. In one model, chickens at day 2 posthatching were used, and in the other, 14-day-old chickens were used. The minimum C. jejuni cell number required to colonize 14-day-old chickens was 5 x 10(4) cells, and that for 2-day-old chickens was 5 x 10(3). The ability of various C. jejuni strains to colonize the chicken gastrointestinal tract was the same in both models.