A method to identify protein antigens of Dermanyssus gallinae for the protection of birds from poultry mites
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The poultry red mite (PRM) Dermanyssus gallinae causes high economic losses and is among the most important parasites in poultry farming worldwide. Different chemical, physical, and biological strategies try to control the expansion of PRM. However, effective solutions to this problem still have to be found. Here, we present a method for the development of an immunological control strategy, based on the identification of mite protein antigens which elicit antibodies with anti-mite activity in the immunized chicken. Hens were immunized with different PRM protein extracts formulated with two different adjuvants, and IgY-antibodies were isolated from the eggs. A PRM in vitro feeding assay which used chicken blood spiked with these IgY-preparations was used to detect antibodies which caused PRM mortality. In vitro feeding of mites with IgY isolated from hens immunized with PRM extract formulated with one of the adjuvants showed a statistically significant increase in the mortality as compared to control mites. After the separation of total PRM extracts in two-dimensional gels, several protein spots were recognized by such IgY preparations. Ten protein spots were subjected to mass spectrometry (MS/MS) for the identification of the corresponding proteins. Complete protein sequences were deduced from genomic and transcriptomic assemblies derived from high throughput sequencing of total PRM DNA and RNA. The results may contribute to the development of an immunological control strategy of D. gallinae.
KeywordsPoultry mite Antigen discovery Vaccine Pest control
We thank Steffen Jakob, Ulrike Ehlert, and Maria Aulmann for the excellent technical assistance. We also thank Drs. Jasmin Fertey and Beyene Moges Agizie for critical reading of the manuscript and support in the statistical analysis. We thank Dr. Daniela Volke (University of Leipzig) for the analysis of PRM 2D gel protein spots.
Compliance with ethical standards
GRM and SU conceived, designed and coordinated the study. GRM performed the experiments, SV carried out the immunizations. MEKJ and MV contributed expertise in birds and poultry experiment design. KS carried out essential sequencing of PRM DNA and RNA. TB performed bioinformatics and statistical analyses. GRM and SU analyzed and interpreted the data and drafted the manuscript. All authors have read and approved the final manuscript.
Conflict of interest
This study was funded by Lohmann Tierzucht GmbH. The funding body had no role in design, collection, analysis, and interpretation of data, in the writing of the manuscript, and in the decision to submit the manuscript for publication.
- Baz M, Samant M, Zekki H, Tribout-Jover P, Plante M, Lanteigne AM, Hamelin M-E, Mallett C, Papadopoulou B, Boivin G (2012) Effects of different adjuvants in the context of intramuscular and intranasal routes on humoral and cellular immune responses induced by detergent-split A/H3N2 influenza vaccines in mice. Clin Vaccine Immunol 19:209–218CrossRefPubMedPubMedCentralGoogle Scholar
- Chabierski S, Makert GR, Kerzhner A, Barzon L, Fiebig P, Liebert UG, Papa A, Richner JM, Niedrig M, Diamond MS, Palù G, Ulbert S (2013) Antibody responses in humans infected with newly emerging strains of West Nile Virus in Europe. PLoS One 8:e66507. doi: 10.1371/journal.pone.0066507 CrossRefPubMedPubMedCentralGoogle Scholar
- Gibb S (2014) MALDIquantForeign: Import/Export routines for MALDIquant. ChicagoGoogle Scholar
- Grabherr MG, Haas BJ, Yassour M, Levin JZ, Thompson D, Amit I, Adiconis X, Fan L, Raychowdhury R, Zeng Q, Chen Z, Mauceli E, Hacohen N, Gnirke A, Rhind N, di Palma F, Birren BW, Nusbaum C, Lindblad-Toh K, Friedman N, Regev A (2011) Full-length transcriptome assembly from RNA-Seq data without a reference genome. Nat Biotechnol 29:644–652CrossRefPubMedPubMedCentralGoogle Scholar
- Haas BJ, Papanicolaou A, Yassour M, Grabherr M, Blood P, Bowden J, Couger M, Eccles D, Li B, Lieber M, MacManes MD, Ott M, Orvis J, Pochet N, Strozzi F, Weeks N, Westerman R, William T, Dewey CN, Henschel R, LeDuc RG, Friedman N, Regev A (2013) De novo transcript sequence reconstruction from RNA-seq using the Trinity platform for reference generation and analysis. Nat Protoc 8:1494–1512CrossRefPubMedGoogle Scholar
- Herbert WJ (1967) Some investigations into the mode of action of the water-in-mineral-oil emulsion antigen adjuvants. In: Symposium Series of Immunobiology Standardization, Karger, Basel, NY:213–220Google Scholar
- Kamau LM, Wright HW, Nisbet AJ, Bowman AS (2013) Development of an RNA-interference procedure for gene knockdown in the poultry red mite, Dermanyssus gallinae: Studies on histamine releasing factor and Cathepsin-D. Afr J Biotechnol 12:1350–1356Google Scholar
- Magnusson SE, Karlsson KH, Reimer JM, Corbach-Söhle S, Patel S, Richner JM, Nowotny N, Barzon L, Bengtsson KL, Ulbert S, Diamond MS, Stertman L (2014) Matrix-M™ adjuvanted envelope protein vaccine protects against lethal lineage 1 and 2 West Nile virus infection in mice. Vaccine 32:800–808. doi: 10.1016/j.vaccine.2013.12.030 CrossRefPubMedGoogle Scholar
- R Core Team (2014) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. URL http://www.R-project.org/
- R Core Team: (2015) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. URL http://www.R-project.org/
- van Emous R (2005) Wage war against the red mite! Poultry Int 44:26–33Google Scholar
- Wickham H (2011) The split-apply-combine strategy for data analysis. J Stat Softw 40:1–29Google Scholar