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  • Author or Editor: George V. Kollias Jr. x
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Abstract

Objectives

To produce monoclonal antibodies (MAB) with specificity for the heavy chain of macaw IgG; to incorporate these MAB into an ELISA to measure IgG responses of macaws inoculated with bovine serum albumin (BSA); and to evaluate the antigenicity of BSA in Blue and Gold Macaws.

Animals

Four 1-year-old Blue and Gold Macaws, 2 males and 2 females.

Procedure

1 male and 1 female 1 were randomly assigned to each of 2 study groups. Group-1 birds were inoculated with 200 μg of BSA on days 0, 14, 28, and 42. Group-2 birds were inoculated with 200 μg of BSA on days 0 and 28. Blood was collected weekly for measurement of anti-BSA titer. Hybridomas were prepared from mice immunized with Scarlet Macaw (Ara macao) IgG purified by salt precipitation and gel chromatography. Specificity for IgG of Scarlet Macaw and other macaw species was confirmed by ELISA and western blot analysis. Hybridoma HL544 was cloned and the antibody purified. Following biotinylation, MAB HL544 was incorporated into an ELISA that measured IgG responses of macaws inoculated with BSA.

Results

Adult Blue and Gold Macaws developed strong primary and secondary anti-BSA antibody titers 14 days after inoculation with 200 μg of BSA. An inoculation interval of 28 days resulted in stronger secondary responses than an interval of only 14 days.

Conclusions

MAB specific for macaw immunoglobulins can be used in ELISA to evaluate the humoral immune responses of macaws. 1-year-old Blue and Gold Macaws developed strong anti-BSA titer when inoculated with 200 μg of BSA. An inoculation interval of 28 days resulted in stronger secondary responses than did an interval of only 14 days.

Clinical Relevance

These MAB, the first reported to have specificity for a psittacine antibody class, will be useful in the evaluation of psittacine antibody responses and in the development of psittacine diagnostic assays. (Am J Vet Res 1996; 57:1157-1161)

Free access
in American Journal of Veterinary Research

Abstract

Objective

To document the maternal transfer of IgG antibodies from Blue and Gold Macaw hens to chicks via the egg; to measure serum IgG half-life in macaw chicks; and to measure the ability of 2- to 10-week-old macaw chicks to generate primary and secondary IgG responses.

Procedure

4 adult Blue and Gold Macaw hens were inoculated with 200 μg of bovine serum albumin (BSA) every 21 days throughout the breeding season. Eggs laid by these hens were incubator hatched to eliminate the possibility of antibody transfer through crop secretions during feeding. Anti-BSA titer was measured just prior to each inoculation in hens and in chicks from 14 to 42 days of age. 1 chick from each of 5 macaw clutches hatched to nonimmunized hens was assigned to 1 of 2 experimental groups. Group-1 chicks were inoculated with 200 μg of BSA at 2 and 6 weeks of age. Group-2 chicks were inoculated with 200 μg of BSA at 6 and 10 weeks of age. Anti-BSA titer was measured weekly for 8 weeks after primary inoculation.

Blood samples were centrifuged, and serum was harvested and frozen at −85 C until analyzed. Anti-BSA IgG titers were measured by ELISA. In the maternal transfer experiment, an exponential decay model was used to calculate the half-life of BSA antibodies in chicks. In the BSA antibody response experiment, comparison of primary and secondary anti-BSA responses of 2- and 6- week-old chicks was performed, using a two-way repeated measures ANOVA, with significance set at P < 0.05.

Results

Hens maintained high anti-BSA titer throughout the breeding season. Maternal transfer of anti-BSA IgG antibodies was documented in all 7 chicks. Anti-BSA titer in chicks decreased in exponential fashion with an average serum IgG half-life of 3.85 days. By 42 days of age, antibodies to BSA were virtually undetectable in all chicks. The primary antibody response of 6-week-old chicks was significantly higher than that of 2-week-old chicks (P = 0.016). No significant difference was observed in the magnitude of the secondary antibody responses between these age groups. Peak anti-BSA IgG antibody responses were reached by 14 days after primary and secondary immunization. Chicks of both age groups generated lower anti-BSA IgG titer than did adult Blue and Gold Macaws.

Conclusions

Blue and Gold Macaw hens transfer IgG antibodies to their chicks through the egg. The half-life of IgG in newly hatched chicks is approximately 3.85 days. 6-week-old chicks develop higher anti-BSA titers than do 2-week-old chicks, but significantly lower titers than do adult macaws.

Clinical Relevance

Information on the nondomestic avian immune system will be useful in the development of vaccination and other preventive health programs for psittacine birds. (Am J Vet Res 1996;57:1162-1167)

Free access
in American Journal of Veterinary Research