Recent studies of Plasmodium falciparum isolated directly from infected patients indicate that alternative parasite biological states occur in the natural host that are not observed with in vitro cultivated parasites. Variation in host substrates, immune responses and other factors probably induce modifications in parasite biology. These biological states could have important implications for pathogenesis, transmission and therapy. We review the differences between P. falciparum in vitro culture systems and in vivo host environments, as well as evidence that host conditions can alter pathogen biology. For select biological questions, the incorporation of naturally occurring conditions into in vitro experimental manipulation of microbes may provide novel insight into pathogen biology.