title: Transposon mutagenesis of Salmonella enterica serovar Enteritidis identifies genes that contribute to invasiveness in human and chicken cells and survival in egg albumen.
abstract: Salmonella enterica serovar Enteritidis is an important food-borne pathogen, and chickens are a primary reservoir of human infection. While most knowledge about Salmonella pathogenesis is based on research conducted on Salmonella enterica serovar Typhimurium, S. Enteritidis is known to have pathobiology specific to chickens that impacts epidemiology in humans. Therefore, more information is needed about S. Enteritidis pathobiology in comparison to that of S. Typhimurium. We used transposon mutagenesis to identify S. Enteritidis virulence genes by assay of invasiveness in human intestinal epithelial (Caco-2) cells and chicken liver (LMH) cells and survival within chicken (HD-11) macrophages as a surrogate marker for virulence. A total of 4,330 transposon insertion mutants of an invasive G1 Nal(r) strain were screened using Caco-2 cells. This led to the identification of attenuating mutations in a total of 33 different loci, many of which include genes previously known to contribute to enteric infection (e.g., Salmonella pathogenicity island 1 [SPI-1], SPI-4, SPI-5, CS54, fliH, fljB, csgB, spvR, and rfbMN) in S. Enteritidis and other Salmonella serovars. Several genes or genomic islands that have not been reported previously (e.g., SPI-14, ksgA, SEN0034, SEN2278, and SEN3503) or that are absent in S. Typhimurium or in most other Salmonella serovars (e.g., pegD, SEN1152, SEN1393, and SEN1966) were also identified. Most mutants with reduced Caco-2 cell invasiveness also showed significantly reduced invasiveness in chicken liver cells and impaired survival in chicken macrophages and in egg albumen. Consequently, these genes may play an important role during infection of the chicken host and also contribute to successful egg contamination by S. Enteritidis.
authors.affiliation: Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, Washington, USA. dshah@vetmed.wsu.edu
authors.forename: Devendra H
authors.initials: DH
authors.lastname: Shah
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Zhou
authors.forename: Hye-Young
authors.initials: HY
authors.lastname: Kim
authors.forename: Douglas R
authors.initials: DR
authors.lastname: Call
authors.forename: Jean
authors.initials: J
authors.lastname: Guard
mesh_terms: D000418:Albumins; D000818:Animals; D018938:Caco-2 Cells; D002645:Chickens / Q000382:microbiology*; D004251:DNA Transposable Elements / Q000235:genetics; D004531:Eggs / Q000382:microbiology*; D004847:Epithelial Cells / Q000382:microbiology*; D005798:Genes, Bacterial / Q000235:genetics*; D044404:Genomic Islands; D006801:Humans; D007422:Intestines / Q000166:cytology / Q000382:microbiology; D008099:Liver / Q000166:cytology / Q000382:microbiology; D016254:Mutagenesis, Insertional / Q000379:methods; D011201:Poultry Diseases / Q000382:microbiology; D012481:Salmonella Infections, Animal / Q000382:microbiology; D012477:Salmonella enteritidis / Q000235:genetics / Q000254:growth & development* / Q000302:isolation & purification / Q000472:pathogenicity*; D014774:Virulence / Q000235:genetics
chemical_list: D000418:Albumins; D004251:DNA Transposable Elements