Production of Coturnix quail immunoglobulins Y (IgYs) against Vibrio parahaemolyticus and Vibrio vulnificus

Abstract

Production of chicken immunoglobulins Y (IgYs) and their applications in prophylactic, therapeutic, detection of microbial contaminants and as a diagnostic tool has been widely studied with limited information from other avians. This study produced Coturnix quail (Coturnix coturnix japonica) egg yolk IgYs against Vibrio parahaemolyticus and Vibrio vulnificus. Formalin inactivated (FIVP, FIVV, mixed FI-VP/VV) and heat inactivated (HIVP, HIVV, mixed HI-VP/VV) Vibrio immunogens (109 CFU/mL) were intramuscularly immunized into quail through thigh muscles. Egg yolk IgY was purified by water dilution-ammonium sulfate precipitation method and the activity was determined by enzyme-linked immunosorbent assay (ELISA). Formalin inactivated immunogens induced high humoral immune response for both V. parahaemolyticus and V. vulnificus over heat inactivated immunogens. However, IgYs resulted from HIVP and FIVV immunogens, showed high specificity to V. parahaemolyticus and V. vulnificus respectively. Detection limits of the indirect ELISA using the produced IgYs were 105 CFU/mL for V. parahaemolyticus and 106 CFU/mL for V. vulnificus. The developed antibodies showed high binding affinity to their corresponding immunogens, very little cross reactivity to Staphylococcus aureus and not other bacteria strains (p<0.05), a phenomenon which was also observed in Western blot.