Dermanyssus gallinae is an economically important blood-feeding ectoparasite affecting layer chicken farms in many countries. Similar to other blood-feeding arthropods, the blood-meal digestion plays a key role in the survival and reproduction of D. gallinae. The knowledge of the genes involved in blood digestion processes may provide new targets for drug and vaccine against the red mites. In the present study, we sequenced and de novo assembled the transcriptomes of unfed and fed adult red mites using Illumina RNA sequencing (RNA-seq) technology. Up to 40,506 unigenes were assembled, and 13,018 unigenes were identified and annotated. GO analysis of the annotated unigenes clustered into three main GO terms. The dominant GO terms of biological processes were cellular process and metabolic process, the prevailing GO terms of the cellular component were cell part and membrane part, and the dominant GO terms of molecular functions were catalytic and binding activities. Up to 6,443 annotated sequences were assigned to 246 active pathways by KEGG analysis. Differentially expressed genes (DEGs) analysis identified 2,877 unigenes with upregulated 2,094 and downregulated 783 in fed female mites compared with unfed female mites. The biological function of these DEGs was further investigated using the KEGG and GO databases. The upregulated DEGs were potentially involved in nutrient metabolism, highlighting their importance in red mite biology. Quantitative reverse transcription real-time PCR (qRT-PCR) validated that the expression levels of the selected six upregulated DEGs were consistent with those in RNA-seq, indicating that the transcriptomic data are reliable. The present study provides valuable and fundamental knowledge that improves our understanding of the physiology of D. gallinae digestion at a molecular level. Moreover, these transcriptomic data will facilitate the identification of novel function genes and candidate antigens for the development of effective vaccines or drug targets to control D. gallinae.Copyright © 2020 Elsevier B.V. All rights reserved.