ABSTRACT
Because of growing levels of antibiotic resistance, new methods to combat bacteria are needed. We hypothesized that because bacteria evolved to survive in specific environments, the addition of compounds, including nutrient type compounds, to an environment, might result in a modification of that environment that will disrupt bacterial growth or in maladaptive bacterial behavior, i.e., gene expression. As a proof of concept, we focused on the egg white environment and the pathogen Salmonella. Despite egg white's antibacterial nature, Salmonella is able to survive and grow in egg white, and this ability of Salmonella leads to infection of chicks and humans. Here, the 20 L-amino-acids were screened for their ability to affect the growth of Salmonella in egg white. L-arginine and L-cysteine were found to reduce growth in egg white in physiologically relevant concentrations. To determine the mechanism behind L-arginine inhibition TnSeq was utilized. TnSeq identified many Salmonella genes required for survival in egg white including genes required for iron import, biotin synthesis, stress responses, cell integrity, and DNA repair. However, a comparison of Salmonella in egg white with and without L-arginine identified only a few differences in the frequency of transposon insertions, including the possible contribution of perturbations in the cell envelope to the inhibition mechanism. Finally, both D-arginine and D-cysteine were found to inhibit Salmonella in egg white. This implied that the effect of arginine and cysteine in egg white is chemical rather than biological, likely on the egg white environment or on the bacterial outer membrane. To conclude, these results show that this approach of addition of compounds, including nutrient type compounds, to an environment can be used to limit bacterial growth. Importantly, these compounds have no inherent anti-bacterial properties, are used as nutrients by animals and bacteria, and only become anti-bacterial in a specific environmental context. Future research screening for the effects of compounds in relevant environments might uncover new ways to reduce pathogen levels in the poultry industry and beyond.
ABSTRACT
The development of probiotics for chickens is a rapidly expanding field. The main approach to probiotics is to administer the probiotic strain throughout the bird's life, usually through incorporation in the feed. However, probiotics which would utilize bacterial strains capable of permanently colonizing the gut after a single exposure are likely to have a greater impact on the developing gut community as well as on the host, would simplify probiotic use and also reduce costs in an industrial setting. Finally, very limited and conflicting information about the colonization ability of different bacterial strains has been reported. Here we report 2 colonization experiments using 14 different bacterial strains from diverse phylogenetic groups. In both experiments, groups of chicks were orally inoculated on the day of hatch with different bacterial strains that had been previously isolated from adult heavy breeders. In the first experiment, colonization of the bacterial strains in broiler chicks was determined 7 d after treatment. In the second experiment, colonization was followed in layer chicks until d 17. Ten of the bacterial strains, including Lactobacillales and Bacteroidales strains, were able to colonize chicks after a single exposure for the duration of the experiment. For a few of these strains, exposure had little effect compared to non-treated chicks due to natural background colonization. Only 4 strains failed to colonize the chicks. Moreover, it is shown that fecal samples are useful to identify and provide a dynamic view of colonization. We further analyzed the effects of artificial colonization on microbiota composition. Some of the strains used in this research were found to reduce Enterobacteriaceae family abundance, implying that they might be useful in reducing relevant pathogen levels. To conclude, our results show that the development of single exposure based probiotics is possible.