The pattern of Campylobacter contamination on broiler farms; external and internal sources.

AIM: The aim of this study was to apply the most sensitive molecular techniques in combination with culture-based methods to characterize broiler farms in terms of the timeline ('appearance' and 'pattern') of Campylobacter contamination prior to and post detection in the birds. METHODS AND RESULTS: Faecal and environmental samples were collected from three broiler farms (two flocks per farm). Real-time PCR was used to test for the presence of Campylobacter. Culture-based methods (enrichment and direct plating) were also applied and isolates were subject to a range of confirmatory tests before speciation (multiplex PCR). All flocks were colonized by Campylobacter before first thin and a similar pattern of Campylobacter contamination was observed; (day -1) a range of external and internal samples real-time PCR positive but culture negative; (day 0) chicks negative; (6-9 days pre-detection in the birds) internal samples (feeders, drinkers, barrier and/or bird weigh) culture positive and (post broiler infection) increasing concentrations of Campylobacter in internal samples but also on the tarmac apron and anteroom. CONCLUSION: It was concluded that; (i) vertical transmission did not occur; (ii) the environment was a potential source of Campylobacter; (iii) testing areas frequented by all birds (e.g. feeders and drinkers), may offer an opportunity for early Campylobacter detection and (iv) once the broilers are infected with Campylobacter, these bacteria are spread from the birds, through the anteroom to the areas surrounding the broiler house, highlighting the need for improved biosecurity. SIGNIFICANCE AND IMPACT OF THE STUDY: This study has established the pattern of Campylobacter contamination on broiler farms, identified an early detection opportunity, highlighted the need to better understand the role of viable but nonculturable Campylobacter in the ecology of Campylobacter on broiler farms and demonstrated the need for improved biosecurity to prevent the spread of Campylobacter from within the house to the surrounding environment.

Quantitative analysis of receptors for adenosine nucleotides obtained via in vitro selection from a library incorporating a cationic nucleotide analog.

5-(3"-Aminopropynyl)-2'-deoxyuridine (dJ), a modified nucleoside with a side chain carrying a cationic functional group, was incorporated into an oligonucleotide library, which was amplified using the Vent DNA polymerase in a polymerase chain reaction (PCR). When coupled to an in vitro selection procedure, PCR amplification generated receptors that bind ATP. This is the first example of an in vitro selection generating oligonucleotide receptors where the oligonucleotide library has incorporated a cationic nucleotide functionality. The selection yielded functionalized receptors having sequences differing from a motif known to arise in a standard selection experiment using only natural nucleotides. Surprisingly, both the natural and the functionalized motifs convergently evolved to bind not one, but two ATP molecules cooperatively. Likewise, the affinity of the receptors for ATP had converged; in both cases, the receptors are half saturated at the 3 mM concentrations of ATP presented during the selection. The convergence of phenotype suggests that the outcome of this selection experiment was determined by features of the environment during which selection occurs, in particular, a highly loaded affinity resin used in the selection step. Further, the convergence of phenotype suggests that the optimal molecular phenotype has been achieved by both selections for the selection conditions. This interplay between environmental conditions demanding a function of a biopolymer and the ability of the biopolymer to deliver that function is strictly analogous to that observed during natural selection, illustrating the nature of life as a self-sustaining chemical system capable of Darwinian evolution.

Redesigning nucleic acids.

A research program has applied the tools of synthetic organic chemistry to systematically modify the structure of DNA and RNA oligonucleotides to learn more about the chemical principles underlying their ability to store and transmit genetic information. Oligonucleotides (as opposed to nucleosides) have long been overlooked by synthetic organic chemists as targets for structural modification. Synthetic chemistry has now yielded oligonucleotides with 12 replicatable letters, modified backbones, and new insight into why Nature chose the oligonucleotide structures that she did.