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Alternatives to Laboratory Animals - ATLA

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

Michael H. Erhard, Katja Mahn,Peter Schmidt, Susanne Oltmer, Rudolf Preisinger, Pia Zinsmeister and Manfred Stangassinger

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.