title: Characterization of the immune response of domestic fowl following immunization with proteins extracted from Dermanyssus gallinae.
abstract: Dermanyssus gallinae is the most significant ectoparasite of European poultry egg laying production systems due to high costs of control and associated production losses as well as adverse effects on bird welfare. In this study, soluble proteins were extracted from unfed D. gallinae (DGE) using a urea-based detergent and ultra-filtration, passed through a 0.22 microm filter and blended aseptically with adjuvant. One group of laying hens was immunized with DGE and adjuvant (Montanide ISA 50 V) whilst another group (Control) received physiological saline and adjuvant. All birds were immunized on two occasions, 21 days apart. Antibody response to immunization was determined by ELISA and western blotting using immunoglobulins (Igs) extracted from egg yolk. DGE immunization of hens resulted in a significant (P<0.05) IgY response compared to controls, although there was no significant difference in IgM response between treatments. A number of proteins were identified by western blotting using IgY antibodies from DGE immunized birds, most prominently at 40 and 230kDa. Analysis of proteins from approximately corresponding bands on SDS-PAGE confirmed the identity of tropomyosin, whilst other proteins showed high sequence homology with myosin and actin from other arachnid and insect species. Immunization of hens with DGE resulted in a 50.6% increase in mite mortality (P<0.001) 17h after feeding when tested by an in vitro mite feeding model. Data in this study demonstrate that somatic antigens from D. gallinae can be used to stimulate a protective immune response in laying hens. Further work is needed to identify other proteins of interest that could confer higher protection against D. gallinae, as well as optimization of the vaccination and in vitro testing protocol.
authors.affiliation: School of Agriculture, Food and Rural Development, Newcastle University, Newcastle upon Tyne, NE1 7RU, UK. daveharrington2004@yahoo.co.uk
authors.forename: David
authors.initials: D
authors.lastname: Harrington
authors.forename: Hatem Mohi El
authors.initials: HM
authors.lastname: Din
authors.forename: Jonathan
authors.initials: J
authors.lastname: Guy
authors.forename: Karen
authors.initials: K
authors.lastname: Robinson
authors.forename: Olivier
authors.initials: O
authors.lastname: Sparagano
mesh_terms: D000818:Animals; D000941:Antigens / Q000276:immunology; D015153:Blotting, Western / Q000662:veterinary; D002645:Chickens; D004530:Egg Yolk / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005247:Feeding Behavior; D007114:Immunization / Q000379:methods / Q000662:veterinary*; D007075:Immunoglobulin M / Q000097:blood / Q000276:immunology; D007136:Immunoglobulins / Q000097:blood / Q000276:immunology*; D019476:Insect Proteins / Q000276:immunology* / Q000494:pharmacology; D008924:Mite Infestations / Q000517:prevention & control / Q000662:veterinary*; D008925:Mites / Q000737:chemistry* / Q000276:immunology; D008970:Molecular Weight; D010316:Particle Size; D011201:Poultry Diseases / Q000517:prevention & control*; D012995:Solubility; D014462:Ultrafiltration / Q000662:veterinary
chemical_list: D000941:Antigens; C016512:IgY; D007075:Immunoglobulin M; D007136:Immunoglobulins; D019476:Insect Proteins