Monitoring of laying capacity, immunoglobulin Y concentration, and antibody titer development in chickens immunized with ricin and botulinum toxins over a two-year period

Abstract

One of the key benefits in using chickens for immunization is the high yield of antibodies obtainable. It is known that egg production decreases over time, while animal maintenance costs remain stable. It would, however, be desirable to keep hens as long as possible to obtain maximal amounts of antibodies. To identify a suitable length of time that animals can be kept and to optimize the cost:yield ratio, we monitored the number of eggs laid, the total amount of chicken IgY, and the specific antibody titer from individually prepared eggs over a 2-yr period. The plant toxin ricin and the Clostridium botulinum neurotoxins type A and B were used to immunize 4 chickens. The number of eggs laid in 2 yr was approximately 600 per hen (about 80% of the maximum egg number), yielding about 20 to 40 g of total IgY per hen. A stable antibody titer of 1:100,000 to 1:1,000,000, as measured by ELISA, was obtained following up to 11 injections of 10 to 20 microg of immobilized native toxin. Laying capacities were found to decrease, on average, from 7 eggs/wk at the point of first immunization to 2 eggs/wk after more than 2 yr. In parallel, the yield of total and specific IgY increased over time, so that the antibody recovery remained high, even after prolonged immunization times. Using purified IgY preparations, classical immunological assays such as ELISA and Western blotting were performed. Furthermore, the IgY showed neutralizing capacity when used to block the functional activity of the toxins both in vitro and in vivo. Analysis of the total IgY content over time demonstrated a complex biological oscillation (and the antigen-specific titer), with a shorter time period of around 7 d (circaseptan rhythm). In summary, we successfully immunized chickens with ricin and botulinum neurotoxins and monitored laying capacity, IgY concentration, and specific antibody titer over an extended period of 2 yr.

Production and characterization of egg yolk antibodies against bovine alimentary tract pathogens

“…In our study the vaccines were given to the hens subcutaneously, whereas majority of former experiments used intramuscular application. Therefore, it is difficult to compare our results to other studies because of different routes of administration of the vaccines, different antigens and adjuvants used as well as different intervals between antigen injections (Schade et al 2005, Pauly et al 2009). Ikemori et al (1992) vaccinated hens with 1 mg of crude pili preparation from E. coli strains 431 (0101:K30; K99; F41.NM,ST) and B44 (09:K30; K99; F41:NM,ST) given i.m.…”

Production and Characterization of IgY against Canine IgG: Prospect of a New Tool for the Immunodiagnostic of Canine Diseases

“…The volume of total IgY isolated from each egg was 2.5 mL, with an average of 97.55 mg of IgY/yolk. Considering the total concentration of specific IgY produced per inoculated antigen was up to 10% (Mine and Kovacs-Nolan 2002;Pauly et al 2009), the maximum concentration of IgY producing an average of 9.75 mg specific IgY would be obtained from one egg yolk. Analyzing the concentration of isolated IgY of each egg yolk collected after the complete immunization, there was a constant profile during all the months, with no significant difference (Kruskal-wallis, H = 2.28, p> 0.05) and without oscillation during the period.…”

“…IgY binds neither to Fc receptors of mammals nor to the mammalian rheumatoid factor, as well as not activating the complement factor, features that reduce interpretation errors, or false positive results (Schade et al 2000). The economic and operational advantages are related to their production, which is continuous and in large quantities throughout the laying period (Schade et al 2000), about two years (Pauly et al 2009). There are also ethical advantages, since the production does not require invasive collection methods, or euthanasia (Schade et al 2000).…”

Evaluation of recombinant Cryptosporidium hominis GP60 protein and anti-GP60 chicken polyclonal IgY for research and diagnostic purposes

“…Production of polyclonal antibodies (IgY class) in laying hens, through extraction of immunoglobulin from egg yolk, is an innovative and expanding branch of biotechnology (PAULY et al, 2009). Replacement of mammalian immunoglobulins by chicken IgY for developing immunological diagnostic tests has several advantages like lower cost, ease of sampling, large quantities of antibodies recovered from egg yolks, high affinity and specificity of antibodies, ease of purification and no need to bleed or sacrifice animals (SCHADE et al, 2005;PAULY et al, 2009).…”

“…Replacement of mammalian immunoglobulins by chicken IgY for developing immunological diagnostic tests has several advantages like lower cost, ease of sampling, large quantities of antibodies recovered from egg yolks, high affinity and specificity of antibodies, ease of purification and no need to bleed or sacrifice animals (SCHADE et al, 2005;PAULY et al, 2009).…”