Effect of norepinephrine on colonisation and systemic spread of Salmonella enterica in infected animals: role of catecholate siderophore precursors and degradation products.

Norepinephrine promotes the growth of Salmonella enterica in vitro in iron-restricted conditions imposed by the iron-binding proteins serum transferrin and egg-white ovotransferrin by facilitating the release of bound iron and subsequent uptake by the bacteria. Moreover, significantly increased colonisation and systemic spread were observed in mouse and chicken models of S. enterica infection following pre-treatment of animals with norepinephrine. Both ent and tonB mutants showed no growth promotion by norepinephrine either in liquid medium containing serum or on plates containing hens' egg-white, indicating that the process is dependent both on the ability to synthesise enterobactin and on TonB-dependent uptake of iron. An entS mutant (formerly designated ybdA) and an iroB mutant behaved as wild type in both assays, showing that neither secretion of enterobactin nor conversion of enterobactin to salmochelin S4 is necessary for the effect. On the other hand, the presence of mutations in fes or iroD resulted in loss of growth promotion by norepinephrine in both assays. Since the fes and iroD genes encode enzymes that hydrolyse enterobactin and salmochelin S4 respectively to monomers, these data suggest that excretion of monomeric forms of these siderophores may be important for the uptake of iron released by norepinephrine from transferrin or ovotransferrin. A similar pattern of behaviour was observed with S. enterica serovar Typhimurium in a mouse model of infection; treatment of animals with norepinephrine before intragastric challenge resulted in increased intestinal colonisation and systemic spread of both wild-type and entS mutant strains, while the fes mutant was significantly attenuated in vivo. This suggests that excretion of 2,3-dihydroxybenzoylserine may be essential for norepinephrine-dependent growth promotion in the iron-restricted environment of the infected host. Unlike the situation in vitro, however, tonB mutants of S. enterica serovars Typhimurium and Enteritidis behaved the same as wild type in mouse and chick infection models, respectively, suggesting that norepinephrine-dependent growth stimulation may also occur by TonB-independent uptake of the enterobactin precursor 2,3-dihydroxybenzoic acid.