[Agglutination of hen egg-yolk immunoglobulins (IgY) against Salmonella enterica, serovar enteritidis]

Two groups of 6 laying hens were used to produce IgY. In the vaccinated group (V), hens were injected by intramuscular route with two doses of a Salmonella enterica serovar Enteritidis bacterin at 20-day interval. In the control group (T) hens remained unvaccinated. Four IgY extractions were performed on the egg production of both groups. The first two extractions were carried out using the yolks obtained from the eggs produced during the 4th and 5th post-vaccination week (extracts 1V and 1T) and the other two using the ones from the 6th, 7th and 8th week (2V and 2T). Starting from the extracts 1V and 1T other products were obtained by freezing-thawing (1V-A and 1T-A) and simple (1V-B and 1T-B) or double (1V-C and 1T-C) flow capillary dialysis concentration. All these products were compared using an ELISA test specific for the detection of chicken antibodies against flagellar antigens of S. Enteritidis. In this test, V extracts were positive whereas T extracts were negative. The extract 1V was more positive than the extract 2V. The extract 1V-C was the most positive and was therefore selected to be used as an antiserum in the agglutination tests. This extract contained 1.9 g/dl of total proteins, 0.028 g/dl of triglycerides and 0.012 g/dl of cholesterol and showed an electrophoretic pattern characteristic of IgY. The 1T-C extract was used as a negative control in the agglutination tests. Slide somatic and tube flagellar agglutination tests were simultaneously carried out using both IgY extracts and a standard rabbit anti-Salmonella (IgG) sera. Overall 367 strains from the Enterobacteriaceae family were tested together with two other strains belonging to the Vibrionaceae family. The 1V-C extract specifically agglutinated S. Enteritidis strains in the same way as the rabbit sera. This extract also agglutinated other Salmonella strains antigenically related to S. Enteritidis. Salmonella which did not share somatic or flagellar antigens with S. Enteritidis, other different species of the Enterobacteriaceae family and the two strains of the Vibrionaceae family were all negative. None of the strains tested was agglutinated by the 1T-C extract. This paper show that it is possible to use specific IgY to identify S. enterica serovars. The more extended use of IgY for diagnostic purposes may be a convenient way to complement the current use of mammal polyclonal antibodies.