Transcriptional profiling of stage specific gene expression in the parasitic ciliate Ichthyophthirius multifiliis

The parasitic ciliate, Ichthyophthirius multifiliis (Ich), is among the most important protozoan pathogens of freshwater fish. Ichthyophthirius cannot be grown in cell culture, and the development of effective prophylactic and therapeutic treatments has been hampered by a lack of information regarding genes involved in virulence, differentiation and growth. To help address this issue, we have generated EST libraries from the two major stages of the parasite life cycle that infect and develop within host tissues. A total of 25,084 ESTs were generated from non-normalized libraries prepared from polyA+ RNA of infective theronts and host-associated trophonts, respectively. Cluster analysis identified 5311 unique transcripts (UniScripts), of which 2091 were contigs and 3220 singletons. Extrapolation of the data based on rates of EST discovery suggests that more than half the expected protein-coding genes of I. multifiliis are represented in this data. BLASTX comparisons against GenBank nr, UniProtKB (SwissProt and TrEMBL), as well as Tetrahymena thermophila, Plasmodium falciparum, and Paramecium tetraurelia protein databases produced 3694 significant (E-value ≤1e−10) hits, of which 1178 were annotated using gene ontology (GO) analysis. A high proportion of UniScripts (63%) showed similarity to other ciliate proteins. When combined with expression profiling data, GO ontology analysis of Biological Process, Cellular Component, and Molecular Function revealed interesting differences in gene families expressed in the two stages. Indeed, the most abundant transcripts were highly stage-specific and coincided with the metabolic activities associated with each stage. This work provides an effective genomics resource to further our understanding of Ichthyophthirius biology, and lays the groundwork for the identification of potential drug targets and vaccines candidates for the control of this devastating fish pathogen.

Host-associated (trophont) and free-swimming (theront) forms of the fish parasite Ichthyophthirius multifiliis show very different patterns of gene expression that reflect the biological and metabolic activities associated with each stage.Figure optionsDownload high-quality image (230 K)Download as PowerPoint slideHighlights
► Comparative gene expression in 2 stages of Ichthyophthirius multifiliis life cycle.
► 25,084 ESTs from free-swimming (theront) and host associated (trophont) stages.
► 5311 unique transcripts identified, 2091 contigs and 3220 singletons.
► Transcriptional profiles differ with developmental stage.
► Abundant transcripts reflect stage-specific metabolic activities.