Authors

Discussion

The poultry red mite (PRM), Dermanyssus gallinae, is the most economically important ectoparasite affecting laying hens throughout the world. Development of alternative control strategies is urgently required, including the development of effective vaccines.  We aim to identify and characterise potential vaccine antigens that are expressed within the gut of the PRM. Starting with homogenised mites, we tested a series of different protein fractionation methods, resulting in the reliable production of a ‘membrane protein enriched’ fraction.  Examination of this fraction by 2D LC-MS resulted in the identification of peptides derived from >1500 contigs of a PRM transcriptome library (Illumina 100bp paired-end sequencing: 36Gb sequence from 36 billion reads: assembled into 200K contigs).  54% of the peptides appear to be shared with other mite species whilst 34% appear to be unique to D. gallinae. The membrane protein enriched fraction was used to select mite-specific antibodies from a phagemid antibody library. We have identified ~400 monoclonal phages of which 16 monoclonals bind specifically to gut sections and will be utilized in future experiments to identify immunogenic gut proteins.