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

Abstract

CONCLUSION:IFA is an intuitionist, sensitive and specific method in detecting S. enteritidis antigen in paraffin wax slices, and it is a good method in diagnosis and antigen location of S. enteritidis . We also conclude that the gland of Garder, heart, kidney, spleen, liver, ileum, jejunum are target organs in S. enteritidis infections of duck, and S. enteritidis is an intracellular parasitic bacterium. INTRODUCTIONSalmonella enteritidis (S. enteritidis) is the infectious disease causing Zoonoses. S. enteritidis is one of the primary causes of human food poisoning throughout the world, and has become a pointed public health problem [1][2][3][4] . China is the biggest country in the raising and consumption of duck in the world, but the S. enteritidis bacillus infection is a severely important infectious disease in the duck industry [5] . S. enteritidis outbreaks have been found to be associated with the consumption of contaminated and undercooked poultry products, such as eggs and egg-containing products, and have become a serious economic and public health problem [6] . Conventional methods for isolation of S. enteritidis are too laborious, not sufficiently sensitive or correct [7,8] . To our knowledge, a serum method for detection and antigen location of S. enteritidis has not been reported. Thus, to establish a rapid, sensitive and highly specific in paraffin slices and antigen location in infected duck tissues. METHODS:Rabbits were immunized with purified bacillus to obtain S. enteritidis -specific antibody, which were then extracted by the caprylic-ammonium sulphate method, purified through High-Q columns. An indirect immuno-fluorescent staining method (IFA) was established to detect the S. enteritidis antigen in paraffin slices. S. enteritidis was detected in each organ tissue of ducklings experimentally infected with S. enteritidis . RESULTS:The gland of Garder, heart, kidney, spleen, liver, brain, ileum, jejunum, bursa of Fabricius from S. enteritidis experimentally infected ducklings were positive or strongly positive, and the S. enteritidis antigen was mainly distributed in the infected cell cytoplasm.

The population of a high-virulence strain of Salmonella enterica serovar Enteritidis in subcutaneously infected partridge: a quantitative time-course study using real-time PCR

“…Severe hyperemia, hemorrhages, and heterophil infiltration in the lungs were also observed. The overall pattern of antigen distribution and microscopic lesions in the organs examined were similar to those reported in previous studies (Akaki et al 1997;Deng et al 2007Deng et al , 2008He et al 2010;Holt and Porter 1992;Yan et al 2008). The factors that determine the levels of S. Enteritidis load in various tissues have not been fully understood; however, the high S. Enteritidis DNA loads in multiple internal organs results in severe tissue pathology, which accelerates the progression of the infection.…”

“…In a previous study, a specific method of indirect immuno-fluorescent antidody staining (IFA) for S. Enteritidis was established (Yan et al 2008). This research based on the IFA assay to study the distribution pattern and quantity for S.Enteritidis in the internal organs of partridge after subcutaneously infected.…”

The Pathogenesis of Salmonella Enteritidis in Experimentally Infected Ducks: A Quantitative Time-Course Study Using TaqMan Polymerase Chain Reaction

“…Severe hyperemia, hemorrhages, and heterophil infiltration in the lungs were also observed. The overall pattern of antigen distribution and microscopic lesions in the organs examined were similar to those reported in previous studies (Holt and Porter, 1992;Akaki et al, 1997;Deng et al, 2008;Yan et al, 2008). The factors that determine the levels of Salmonella Enteritidis load in various tissues have not been fully understood; however, the high Salmonella Enteritidis DNA loads in multiple systemic organs results in severe tissue pathology, which accelerates the progression of the infection.…”

Detection and localization of a goose adenovirus in experimentally infected goslings, using indirect immunofluorescence with paraffin-embedded tissue sections

“…Based on previous studies (Smith et al, 1991;Bertsy et al, 2008;Yan et al, 2008), the following factors were considered. (1) Antigen recovery, to partly recover the antigen based on a microwave using citrate buffer solution (0.01 M, pH 6.0) for 20 min, parenzyme treatment (200 mg/ml) for 20 min or no recovery, respectively.…”