Tubular glands of the isthmus are the predominant colonization site of Salmonella enteritidis in the upper oviduct of laying hens

Department of Pathology, Bacteriology and Avian Diseases, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium. jeroen.debuck@UGent.be

Salmonella enterica serovar Enteritidis is the serovar most frequently isolated from chicken eggs. Colonization of the upper oviduct of hens is believed to play an important role in egg contamination. The interaction of S. enteritidis with gland epithelial cells of the isthmus and the magnum was, therefore, studied in vitro and in vivo. In the first experiment, S. enteritidis bacteria were added to confluent monolayers of primary cultures of chicken tubular epithelial cells of the isthmus (ICTEC) or magnum (MCTEC). Intracellular bacteria in ICTEC and MCTEC were confirmed by a gentamicin protection assay. Internalization in the glandular cells was corroborated by confocal scanning microscopy. Although S. enteritidis was able to invade and proliferate intracellularly during 24 h in the cell culture of both segments, this was significantly more so in the ICTEC. In a second experiment, an in vivo loop model was developed for investigation of the invasiveness of S. enteritidis in the oviduct of laying hens. Loops in the isthmus and magnum of laying hens were made under anaesthesia. S. enteritidis was inoculated into each loop. After 1 h, tubular gland cells were isolated from the infected tissue under gentamicin. S. enteritidis invasiveness was measured as the ratio of isolated bacteria per isolated tubular gland cell. This ratio was higher (P < 0.01) in the isthmus (1.3 x 10(-3)) than in the magnum (5.3 x 10(-5)). In a third experiment, laying hens were intravenously infected with 5 x 10(7) cfu S. enteritidis bacteria. The number of intracellular bacteria was counted in the tubular gland cells of the isthmus and magnum after isolation under gentamicin. The ratio of isolated bacteria per isolated tubular gland cell was again significantly higher in the isthmus as compared with in the magnum. In all 3 assays, the tubular gland cells of the isthmus were more heavily invaded than those of the magnum.