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Background  Multiple infections of the same host by different strains of the same microparasite species are believed to play a crucial role during the evolution of parasite virulence. We investigated the role of specificity, relative virulence and relative dose in determining the competitive outcome of multiple infections in the  Daphnia magna - Pasteuria ramosa  host-parasite system.    Results  We found that infections by  P. ramosa  clones (single genotype) were less virulent and produced more spores than infections by  P. ramosa  isolates (possibly containing multiple genotypes). We also found that two similarly virulent isolates of  P. ramosa  differed considerably in their within-host competitiveness and their effects on host offspring production when faced with coinfecting  P. ramosa  isolates and clones. Although the relative virulence of a  P. ramosa  isolate/clone appears to be a good indicator of its competitiveness during multiple infections, the relative dose may alter the competitive outcome. Moreover, spore counts on day 20 post-infection indicate that the competitive outcome is largely decided early in the parasite’s growth phase, possibly mediated by direct interference or apparent competition.    Conclusions  Our results emphasize the importance of epidemiology as well as of various parasite traits in determining the outcome of within-host competition. Incorporating realistic epidemiological and ecological conditions when testing theoretical models of multiple infections, as well as using a wider range of host and parasite genotypes, will enable us to better understand the course of virulence evolution.

The expression and evolution of virulence in multiple infections: the role of specificity, relative virulence and relative dose