Isolation and Characterization of IgM and IgY Antibodies from Plasma of Magellanic Penguins (Spheniscus magellanicus)

Abstract

Infectious diseases such as aspergillosis, avian malaria, and viral infections are significant threats to the conservation of penguins, leading to morbidity and mortality of these birds both in captivity and in the wild. The immune response to such infectious diseases is dependent on different mechanisms mediated by cells and soluble components such as antibodies. Antibodies or immunoglobulins are glycoproteins that have many structural and functional features that mediate distinct effector immune functions. Three distinct classes of antibodies have been identified in birds: immunoglobulin A (IgA), immunoglobulin M (IgM), and immunoglobulin Y (IgY). In this study we aim to establish an efficient laboratory method to obtain IgM and IgY antibodies from plasma samples of healthy adult Magellanic penguins (Spheniscus magellanicus). The protocol was developed combining plasma delipidation, sequential precipitation with caprylic acid and ammonium sulfate, and size-exclusion chromatography. The efficiency of the protocol and the identity of the purified IgM and IgY antibodies were confirmed through enzyme-linked immunosorbent assay, Western blotting, one-dimensional and two-dimensional polyacrylamide gel electrophoresis, and lectin binding assay. Structural and physicochemical properties of IgM and IgY from Magellanic penguins were consistent with those of other avian species. This purification protocol will allow for more detailed studies on the humoral immunity of penguins and for the development of high specificity serologic assays to test Magellanic penguins for infectious pathogens.

Health evaluation of African penguins (<i>Spheniscus demersus</i>) in southern Africa

“…While these specific commercial tests have not undergone thorough validation to estimate their sensitivity and specificity when applied to samples from African penguins, other studies on the antigenic properties of penguin immunoglobulins corroborate the validity of their basic methodological assumption (Bizelli et al . 2015 ; Graczyk et al . 1994 , 1995 ).…”

“…2004 ), which hopefully will be overcome through ongoing research aiming at the production of secondary antibodies specifically targeting penguin IgY (Bizelli et al . 2015 ). On the other hand, the HIA used to test for NDV, AIV H5 and AIV H7 is not subject to this limitation because it does not rely on the recognition by secondary antibodies.…”

Application of chimeric antigens to paper-based diagnostics for detection of West Nile virus infections ofCrocodylus porosus –a novel animal test case

“…To determine the suitability of the goat anti-alligator IgY conjugate for detection of saltwater crocodile antibodies, size exclusion chromatography purified crocodile IgY (Bizelli et al 2015) was striped onto nitrocellulose membrane at a concentration of 1 mg/mL and dried at 37℃ for 1 hour. The anti-alligator IgY conjugate was applied to conjugate pads and dried.…”