Development and application of an ELISA for the detection of duck antibodies against Riemerella anatipestifer antigens in egg yolk of vaccinees and in serum of their offspring

Abstract

A direct and an indirect antibody enzyme-linked immunosorbent assay for duck yolk IgY and duck serum IgY was developed and tested on egg yolk and serum of ducks vaccinated with Riemerella anatipestifer (Ra). Tests were performed either with primary antibodies labelled with horseradish peroxidase or with alkaline phosphatase-labelled secondary antibodies reacting with specifically bound rabbit anti-duck IgY antibodies, respectively. Ra-specific IgYs in egg yolk from three ducks increased rapidly at day 8 after the first of two vaccinations. In two ducks, the IgY titre persisted on a high plateau for 3 months. The concentration of Ra-specific IgYs in the serum of the progeny of vaccinees decreased between day 3 and day 10 after hatching. The fraction of total IgYs decreased less but also significantly. It was shown that antibodies were vertically transmitted and therefore protect offspring against Ra infection at least during the first week after hatching. The test design with anti-IgY rabbit antibodies is further suitable to detect other specific antibodies if respective antigens were fixed on solid phases.

Pathogenesis ofRiemerella anatipestiferin turkeys after experimental mono-infection via respiratory routes or dual infection together with the avian metapneumovirus

“…Serum antibody response to RA was examined by an IgG-ELISA system following protocols previously developed for detection of RA antibodies in ducks (Hatfield et al, 1987;Lobbedey & Schlatterer, 2003). In contrast to the published protocol, we used commercially available dilution buffer, ABTS substrate, and stop solution and the anti-turkey-IgG conjugate instead of anti-duck-IgG.…”

Characterization of specific egg yolk immunoglobulin (IgY) against mastitis-causingStaphylococcus aureus

“…The specific activity of IgY against Staph. aureus was tested by indirect ELISA (Lobbedey and Schlatterer 2003). The WSF from immunized hens were diluted in a doubling series from 1 : 100 to 1 : 51 200 in phosphate‐buffered saline (PBS, pH 7·4).…”

Effects of specific egg yolk immunoglobulin on pan-drug-resistant Acinetobacter baumannii

“…Titer of IgYs specific to Ab-B (B-IgY) and Ab-C (C-IgY) was determined by enzyme-linked immunosorbent assay (ELISA) conducted on WSF fractions [27]. Microtiter plate was coated with Ab-B or Ab-C at a concentration of 10 9 cfu/ml.…”