A key bacterial virulence factor is the ability to acquire the micronutrient iron, required by a vast majority of micro-organisms for survival and proliferation within their hosts. The work described here focuses on acquisition of iron in Pasteurella multocida serotype A:3, a Gram-negative opportunistic pathogen that causes pneumonic pasteurellosis in cattle, a severe respiratory infection of significant economic burden and welfare concern. P. multocida A:3 acquires iron primarily from the host iron-chelating molecule transferrin through the expression of specific outer membrane receptors. The correlation between up-regulation of these receptors and of other iron-regulated outer membrane proteins (IROMPs) and an increase in bacterial virulence has been noted elsewhere. To date, there has been no systems analysis of the metabolic processes of iron acquisition published for any bacterial species. The work described here used a systems approach involving elementary flux modes analysis to derive a computational model of iron acquisition and reveal a number of key findings on the mechanisms of iron acquisition and links between iron uptake, glucose metabolism and protein synthesis. This in silico model was subjected to experimental validation through a range of in vitro experiments designed to investigate the links between iron restriction and growth and metabolism of P. multocida. This novel approach was only possible after the development and optimisation of a number of assays to measure key model parameters.