Evaluation of Various Immunisation Procedures in Laying Hens to Induce High Amounts of Specific Egg Yolk Antibodies

Abstract

The present study, involving 972 laying hens divided into 162 groups (n = 6), was aimed at the development of an immunisation protocol for laying hens to produce specific egg yolk antibodies. Recombinant bovine somatotropin (rbst), Escherichia coli pilus antigen K88 (K88), human serum immunoglobulin G (IgG), and low density lipoprotein (LDL) were used as antigens, each at four different doses (rbst, K88, LDL: 1μg, 10μg, 100μg, 1mg; IgG: 0.5μg, 5μg, 50μg, 0.5mg). Three subcutaneous or intramuscular immunisations were performed at intervals of four weeks. The adjuvant used was either the lipopeptide Pam3Cys-Ser-(Lys)4 (PCSL) or Freund's incomplete adjuvant (FIA), in two different doses (PCSL: 0.1 and 0.25mg; FIA: 0.1 and 0.25ml). In the four antigen control groups, hens were immunised without any adjuvant. In two negative control groups, only physiological saline was injected. The mean egg weight and egg yield were not influenced by the immunisation procedures. An antigen dose of 10–100g per injection was sufficient to induce high specific antibody titres in the egg yolk. The adjuvant efficacy of PCSL and FIA was proved to be the same (p < 0.05 versus antigen control). With PCSL as adjuvant, some groups showed a tendency to produce even higher specific antibody titres than did FIA groups. A second booster often caused a further significant increase in the amounts of specific antibodies, especially with PCSL. Subcutaneous administration of the antigen together with 250μg PCSL, resulted in a significantly higher immune response than when FIA was used.

Raising Antibodies in Chickens Against Primaquine, Pyrimethamine, Dapsone, Tetracycline, and Doxycycline

“…Chickens have successfully been used to generate antibodies against a range of antigens and due to the phylogenetic distance between chicken and mammalian antibodies (Erhard et al, 2000;Larsson and Sjoquist, 1990); chicken antibodies have reduced cross-reactivity with mammalian proteins and do not activate mammalian complement (Carlander and Larsson, 2001). Chickens produce high titres of antibodies (Schade et al, 2001) and antibodies can be collected from chicken eggs, obviating the need for invasive bleeding techniques required for harvesting antibodies in other animals.…”

Critical Steps in the Production of Polyclonal and Monoclonal Antibodies: Evaluation and Recommendations

“…When larger amounts of PAbs are needed, farm animals such as sheep, goats, and horses are usually used. In some cases requiring large amounts of PAbs, chickens may be used (Erhard et al 2000;Schade et al 1996). After the immunization of chickens, antibodies pass from the blood to the egg yolk.…”

“…After the immunization of chickens, antibodies pass from the blood to the egg yolk. Chicken egg antibodies (IgY) can be extracted from the egg to concentrations of approximately 100 to 250 mg of IgY/egg (CCAC 2002;Erhard et al 2000). Compared with the IgG productivity in rabbits (approximately 250 mg per bleed), more IgY can be obtained from chickens due to the continuous secretion of IgY in the eggs.…”

The Humoral Immune Response and the Productivity of Laying Hens Kept on the Ground or in Cages

“…In each of the four groups, the immune response to human IgG was determined in the yolk samples of eggs, collected once a week. The concentrations of specific chicken IgY (100 ELISA units correspond to 0.81mg human IgG specific IgY) and of total IgY in the egg yolk samples were measured with ELISA systems, as described by Erhard et al (5,6).…”

“…The 1996 workshop in Berlin, organised by the European Centre for the Validation of Alternative Methods (ECVAM), summarised the state of the art of production of avian egg yolk antibodies (2). As described recently (2)(3)(4)(5), the production of antibodies in chickens is now internationally accepted as a refinement method. The number of animals used for such purposes can thus be significantly reduced, because of the high amounts of antibody produced (up to 250mg IgY per egg yolk).…”