Immunoglobulin detection in wild birds: effectiveness of three secondary anti‐avian IgY antibodies in direct ELISAs in 41 avian species

Abstract

Summary Immunological reagents for wild, non-model species are limited or often non-existent for many species.In this study, we compare the reactivity of a new anti-passerine IgY secondary antibody with existing secondary antibodies developed for use with birds. Samples from 41 species from the following six avian orders were analysed: Anseriformes (1 family, 1 species), Columbiformes (1 family, 2 species), Galliformes (1 family, 1 species), Passeriformes (16 families, 34 species), Piciformes (1 family, 2 species) and Suliformes (1 family, 1 species). Direct ELISAs were performed to detect total IgY using goat anti-passerine IgY, goat anti-chicken IgY or goat anti-bird IgY secondary antibodies.The anti-passerine antibody exhibited significantly higher IgY reactivity compared to the anti-chicken and/or anti-bird antibodies in 80% of the passerine families tested. Birds in the order Piciformes (woodpeckers) and order Suliformes (cormorants) were poorly detected by all three secondary antibodies. A comparison of serum and plasma IgY levels was made within the same individuals for two passerine species (house finch and white-crowned sparrow), and serum exhibited significantly more IgY than the plasma for all three secondary antibodies. This result indicates that serum may be preferred to plasma when measuring total antibody levels in blood.This study indicates that the anti-passerine IgY secondary antibody can effectively be used in immunological assays to detect passerine IgY for species in most passerine families and is preferred over anti-chicken and anti-bird secondary antibodies for the majority of passerine species. This anti-passerine antibody will allow for more accurate detection and quantification of IgY in more wild bird species than was possible with previously available secondary antibodies.

Host tolerance and resistance to parasitic nest flies differs between two wild bird species

“…One remaining question is why are swallows more immunologically resistant to the parasite compared to bluebirds? Fassbinder‐Orth et al () found that antibodies can bind with different affinity to detection antibodies, and therefore, it is possible that bluebird antibodies have a low affinity to the detection antibody used in our study. However, a recent study found that bluebird females and nestlings supplemented with mealworms do produce an IgY response (i.e., have similarly high OD values) (Knutie, ), and therefore, detection antibody binding is likely similar across species.…”

Immunosenescence in wild animals: meta‐analysis and outlook

“…Our analysis indicated that the abundance of naïve cells are also strongly sensitive to age in wild animals, and thus it seems promising to adapt flow cytometry assessment of memory/naïve cell subtypes (see Table ); this has so far been done for monkeys and bovids (see Table ), but such ‘immunophenotyping’ techniques are also available for birds (Kaiser ), although there can be some difficulty in identifying unequivocal markers for naïve cells (Müller et al ). Measurements of circulating memory antibodies are not technically challenging, and recent development of a passerine specific anti‐IgY, in addition to a bird and chicken‐specific antibody (Fassbinder‐Orth et al ), is potentially a useful addition to the toolkit of avian immune ecologists. Within the innate system, reduced neutrophil and NK cell activity are predictive of increased mortality in old humans, while dysregulation of TLR function affects responsiveness to viral infections and vaccines (see Table ; review in Panda et al ), which can be measured in ecological studies (an example in birds: Martin et al ).…”

<p>Comparison Of Four Anti-Avian IgY Secondary Antibodies Used In Western Blot And Dot-Blot ELISA To Detect Avian Bornavirus Antibodies In Four Different Bird Species</p>

“…Though some studies have used antichicken secondary antibody in serologic assays in wild birds, 28,29 our results were in agreement with a study that found the use of anti-passerine IgY secondary antibody in a microplate ELISA performed better than anti-chicken and anti-bird secondary antibodies when testing passerines. 25 In fact, we found that the use of antimacaw IgY secondary antibody, in both the Western blot and the dot-blot ELISAs, showed strong species specificity for psittacines. Conversely, the use of anti-macaw IgY secondary antibody showed no difference between positive and negative Mallard plasma in either of the immunological assays (Figures 1 and 2; Table 1).…”

“…Anti-passerine IgY secondary antibody produces better results than the anti-bird IgY secondary antibody or the anti-chicken IgY secondary antibody for serologic assays on passerine birds. 25 This suggests that species-specific secondary antibodies may provide more sensitive results in immunologic assays than commercially available mixed species anti-bird secondary antibody. In assays that employ short antigen-antibody incubation times, such as dot-blot or lateral flow ELISAs, a species-specific secondary antibody may be more useful when testing psittacine birds.…”