The increasing application of RNA-seq to study non-model organisms demands easy-to-use and efficient bioinformatics tools to help researchers quickly uncover biological and functional insights from large datasets. Here, we present a unified software suite for processing, analyzing, and interpreting RNA-seq data from any eukaryotic species. This suite consists of a) EcoOmicsDB (www.ecoomicsdb.ca), a database for ortholog mapping and cross-species comparison; b) EcoOmicsAnalyst (www.ecoomicsanalyst.ca), a platform for raw data processing and annotation; and c) ExpressAnalyst (www.expressanalyst.ca), a platform for statistical and functional analysis. The utilities of this suite are demonstrated through case studies of RNA-seq data from multiple non-model species with or without reference transcriptomes. By coupling ultra-fast read mapping algorithms with high-resolution ortholog databases through a user-friendly web interface, the tool suite enables researchers to obtain global expression profiles and gene-level insights from raw RNA-seq reads within 24 hours.

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Chemical risk assessment for avian species typically depends on information from toxicity tests performed in adult birds. Early-life stage (ELS) toxicity tests have been proposed as an alternative, but incorporation of these data into existing frameworks will require knowledge about the similarities/differences between ELS and adult responses. The present study uses transcriptomics to assess hepatic gene expression in ELS and adult Japanese quail following exposure to ethinylestradiol (EE2). Prior to incubation, ELS quail were dosed with measured EE2 concentrations of 0.54, 6.3, and 54.2 µg/g egg weight via air cell injection. Adult quail were fed a single dose of EE2 at nominal concentrations of 0, 0.5, and 5 mg/kg body weight by gavage. Liver tissue was collected from five to six individuals per dose group at mid-incubation for ELS quail and 4 days after dosing for adults. A total of 283 and 111 differentially expressed genes (DEGs) were detected in ELS and adult quail, respectively, 16 of which were shared across life stages. Shared DEGs included estrogenic biomarkers such as vitellogenin genes and apovitellenin-1. For the dose groups that resulted in the highest number of DEGs (ELS, 6.3 µg/g; adult, 5 mg/kg), 21 and 35 Kyoto Encyclopedia of Genes and Genomes pathways were enriched, respectively. Ten of these pathways were shared between life stages, including pathways involved with signaling molecules and interaction and the endocrine system. Taken together, our results suggest conserved mechanisms of action following estrogenic exposure across two life stages, with evidence from differential expression of key biomarker genes and enriched pathways. The present study contributes to the development and evaluation of ELS tests and toxicogenomic approaches and highlights their combined potential for screening estrogenic chemicals.

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