Next Generation Sequencing of Newly excysted juveniles, Juveniles and Adult Fasciola hepatica

Purpose: The miRnome of the liver fluke, Fasciola hepatica, has historically been assembled with limited omic's resources using specific life stages. The goals of this study is to quanity the known and published miRNAs and determine novel miRNAs across three intra-mammlian life stages. Methods: Total RNA was extracted from newly ecysted juveniles 24h post excystment, juveniles at 21d post infection in rats and adult worms during hepatic infection in sheep. Total RNA of each sample was prepared for miRNA sequencing library. Sequenced reads were cleaned and quanitified for F. hepatica mature miRNAs from miRBase version 21 and published miRNAs from Ricafrente et al 2021 using Bowtie tool. Cleaned reads were additionally analysed for novel miRNAs using MiRDeep2 tool in conjunction with the F. hepatica genome and mature miRNAs from F. hepatica, S. japonicum, S. mansoni and C.elegans. Known/published and novel miRNA read counts were normalised to CPM. Results: Of all miRNAs known/published and novel, 124 miRNAs now make up the F. hepatica miRnome. Each life stage exhibited a unique miRNA profile, from which NEJs were the most differentially expressed compared to juveniles and adult. Conclusions: Our study is the first to comparatively assess the miRNA profiles of the intrammalian life stages of F.hepatica simultaneously, from which the miRnome has now been expanded from 77 to 124. miRNA profiles of Fasciola hepatica life stages; newly excysted juveniles 24h, juveniles 21d and adult