Evaluation of SEF14 fimbrial dot blot and flagellar Western blot tests as indicators of Salmonella enteritidis infection in chickens

Abstract

The serological responses to Salmonella enteritidis flagella (H: g,m) and its fimbrial antigen SEF14 were evaluated as indicators of infection in chickens and to confirm serological results obtained by an ELISA using S enteritidis lipopolysaccharide (LPS) (O: 9,12) as the detecting antigen. The SEF14 antigen and flagella were extracted from S enteritidis and transferred to nitrocellulose paper for use in Western and dot blot tests. Antisera to 19 salmonella serotypes including S enteritidis were raised in rabbits and their cross reactivity to the flagellar and SEF14 antigens was evaluated. Cross reactivity with the SEF14 antigen was found in one antiserum, raised against S blegdam, and to flagella in eight of 19 antisera raised against various salmonella serotypes, most of which shared the flagellar factors g or m with S enteritidis. The intensity of cross reaction to flagella was strongest in S derby and S blegdam antisera. Antisera raised in chickens against S typhimurium and S panama did not cross react in either test, and neither did pooled sera from eight-week-old salmonella-free, broiler breeder parent chickens. Field sera from two commercial flocks with no history of salmonella infection were negative when tested by the LPS ELISA. These sera were also negative when tested by the flagellar and SEF14 blots. S enteritidis infection in a commercial laying flock was detected initially when the sera were tested by the LPS ELISA and confirmed in individual and pooled sera by the SEF14 and flagellar tests. S enteritidis PT4 was isolated from this flock post mortem.

Application of an indirect immunofluorescent staining method for detection of Salmonella enteritidis in paraffin slices and antigen location in infected duck tissues

“…Desmidt et al [14] observed the S. enteritidis in the ceca of chickens by immunohistochemistry, which can be used to locate subcells of organ tissues of S. enteritidis infected chickens. Coope [15] detected the S . enteritidis by Dot-Blot and Western-Blot, which is rapid and specific, but could not detect the location in sub-cells of organ tissues.…”

Igg Subclasses Targeting the Flagella of Salmonella Enterica Serovar Typhimurium Can Mediate Phagocytosis and Bacterial Killing

“…Therefore functional data on the effect of antibody binding to candidate surface antigens are essential for preclinical vaccine development. The Salmonella flagellum is a main bacterial target, inducing efficient antibody production upon infection [25][26][27]. Due to its surface localization, repetitive epitopes along their structure, and TLR-5-associated pro-inflammatory adjuvanticity, flagella can be considered as good protein carriers for inclusion in conjugate vaccines against iNTS.…”

“…Infection with S. enterica induces production of antibodies against various bacterial targets such as flagellar proteins, outer membrane proteins, lipopolysaccharide (LPS), heat shock proteins and fimbriae [24][25][26][27][28]. However, the full spectrum of the antigen specificity of the protective antibody response against invasive salmonelloses is still unclear.…”

Humoral immune response in hens naturally infected withSalmonellaEnteritidis against outer membrane proteins and other surface structural antigens

“…These cross-reactions were not observed in the case of flagellin and fimbriae, indicating the specificity of the antibodies generated against these surface components during an infection by S. Enteritidis in hens. In fact, different authors have taken advantage of this property employing these components individually for the development of specific serodiagnostic tests, like SEF14 and flagellin in ELISA [2,5,11,24,29,33].…”