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Background  Third-stage larvae (L3) of the canine hookworm,  Ancylostoma caninum , undergo arrested development preceding transmission to a host. Many of the mRNAs up-regulated at this stage are likely to encode proteins that facilitate the transition from a free-living to a parasitic larva. The initial phase of mammalian host invasion by  A. caninum  L3 (herein termed “activation”) can be mimicked  in vitro  by culturing L3 in serum-containing medium.    Methodology/Principal Findings  The mRNAs differentially transcribed between activated and non-activated L3 were identified by suppression subtractive hybridisation (SSH). The analysis of these mRNAs on a custom oligonucleotide microarray printed with the SSH expressed sequence tags (ESTs) and publicly available  A .  caninum  ESTs (non-subtracted) yielded 602 differentially expressed mRNAs, of which the most highly represented sequences encoded members of the pathogenesis-related protein (PRP) superfamily and proteases. Comparison of these  A. caninum  mRNAs with those of  Caenorhabditis elegans  larvae exiting from developmental (dauer) arrest demonstrated unexpectedly large differences in gene ontology profiles.  C .  elegans  dauer exiting L3 up-regulated expression of mostly intracellular molecules involved in growth and development. Such mRNAs are virtually absent from activated hookworm larvae, and instead are over-represented by mRNAs encoding extracellular proteins with putative roles in host-parasite interactions.    Conclusions/Significance  Although this should not invalidate  C .  elegans  dauer exit as a model for hookworm activation, it highlights the limitations of this free-living nematode as a model organism for the transition of nematode larvae from a free-living to a parasitic state.

Transcriptional Changes in the Hookworm, Ancylostoma caninum, during the Transition from a Free-Living to a Parasitic Larva