Molecular Diagnosis & Therapy

, Volume 17, Issue 5, pp 311–317 | Cite as

Distribution of Six Effector Protein Virulence Genes Among Salmonella enterica enterica Serovars Isolated from Children and their Correlation with Biofilm Formation and Antimicrobial Resistance

  • A. Ioannidis
  • K. Papavasileiou
  • E. Papavasileiou
  • S. Bersimis
  • S. Chatzipanagiotou
Original Research Article



Salmonella enterica enterica encodes a variety of virulence factors. Among them, the type III secretion system (TTSS) encoded in the Salmonella pathogenicity islands (SPIs) is required for induction of proinflammatory responses, invasion of intestinal epithelial cells, induction of cell death in macrophages, and elicitation of diarrhea. The presence of the effector protein genes sopB, sopD, sopE, sopE2, avrA, and sptP of the SPIs was analyzed in 194 S. enterica enterica strains belonging to 19 serovars.


S. enterica enterica strains were collected from children with gastroenteritis, either hospitalized or attending the outpatient clinic, aged 1-14 years. Nineteen different serotypes were included in the study. Serotyping, biofilm formation determination, and antimicrobial resistance of the planktonic as well as the biofilm forms of the strains have been reported previously.


At least one virulence gene was present in all Salmonella isolates. Biofilm formation was statistically independent of any of the six genes. Strains lacking sopE and sopE2 were more resistant to all the antimicrobials.


The association of the virulence genes with the antimicrobial resistance of Salmonella in general has been previously reported and is a matter of further investigation. For the clinical expression of pathogenicity in humans, the contribution of these genes is questionable, as some strains bearing only a single gene (either sptP or avrA) were still capable of causing gastroenteritis.


Virulence Gene Gastroenteritis Antimicrobial Resistance Moxifloxacin Antimicrobial Susceptibility 



This study was financially supported by the Aeginition Hospital of the Athens Medical School (Athens, Greece). The authors have no conflicts of interest that are directly relevant to the content of this study.


  1. 1.
    Mirold S, Ehrbar K, Weissmuller A, Prager R, Tschape H, Russmann H, et al. Salmonella host cell invasion emerged by acquisition of a mosaic of separate genetic elements, including Salmonella pathogenicity island 1 (SPI1), SPI5, and sopE2. J Bacteriol. 2001;183(7):2348–58.PubMedCrossRefGoogle Scholar
  2. 2.
    Fluit AC. Towards more virulent and antibiotic-resistant Salmonella? FEMS Immunol Med Microbiol. 2005;43(1):1–11.PubMedCrossRefGoogle Scholar
  3. 3.
    Groisman EA, Ochman H. Pathogenicity islands: bacterial evolution in quantum leaps. Cell. 1996;87(5):791–4.PubMedCrossRefGoogle Scholar
  4. 4.
    Ehrbar K, Friebel A, Miller SI, Hardt WD. Role of the Salmonella pathogenicity island 1 (SPI-1) protein InvB in type III secretion of SopE and SopE2, two Salmonella effector proteins encoded outside of SPI-1. J Bacteriol. 2003;185(23):6950–67.PubMedCrossRefGoogle Scholar
  5. 5.
    Galan JE. Salmonella interactions with host cells: type III secretion at work. Annu Rev Cell Dev Biol. 2001;17:53–86.PubMedCrossRefGoogle Scholar
  6. 6.
    Santos RL, Zhang S, Tsolis RM, Kingsley RA, Adams LG, Baumler AJ. Animal models of Salmonella infections: enteritis versus typhoid fever. Microbes Infect. 2001;3(14–15):1335–44.PubMedCrossRefGoogle Scholar
  7. 7.
    Wallis TS, Galyov EE. Molecular basis of Salmonella-induced enteritis. Mol Microbiol. 2000;36(5):997–1005.PubMedCrossRefGoogle Scholar
  8. 8.
    Bakshi CS, Singh VP, Wood MW, Jones PW, Wallis TS, Galyov EE. Identification of SopE2, a Salmonella secreted protein which is highly homologous to SopE and involved in bacterial invasion of epithelial cells. J Bacteriol. 2000;182(8):2341–4.PubMedCrossRefGoogle Scholar
  9. 9.
    Hansen-Wester I, Hensel M. Salmonella pathogenicity islands encoding type III secretion systems. Microbes Infect. 2001;3(7):549–59.PubMedCrossRefGoogle Scholar
  10. 10.
    Jennings ME, Quick LN, Ubol N, Shrom S, Dollahon N, Wilson JW. Characterization of Salmonella type III secretion hyper-activity which results in biofilm-like cell aggregation. PLoS One. 2012;7(3):e33080.PubMedCrossRefGoogle Scholar
  11. 11.
    Parvathi A, Vijayan J, Murali G, Chandran P. Comparative virulence genotyping and antimicrobial susceptibility profiling of environmental and clinical Salmonella enterica from Cochin, India. Curr Microbiol. 2011;62(1):21–6.PubMedCrossRefGoogle Scholar
  12. 12.
    Dione MM, Ikumapayi U, Saha D, Mohammed NI, Adegbola RA, Geerts S, et al. Antimicrobial resistance and virulence genes of non-typhoidal Salmonella isolates in The Gambia and Senegal. J Infect Dev Ctries. 2011;5(11):765–75.PubMedGoogle Scholar
  13. 13.
    Hapfelmeier S, Ehrbar K, Stecher B, Barthel M, Kremer M, Hardt WD. Role of the Salmonella pathogenicity island 1 effector proteins SipA, SopB, SopE, and SopE2 in Salmonella enterica subspecies 1 serovar Typhimurium colitis in streptomycin-pretreated mice. Infect Immun. 2004;72(2):795–809.PubMedCrossRefGoogle Scholar
  14. 14.
    Miko A, Pries K, Schroeter A, Helmuth R. Molecular mechanisms of resistance in multidrug-resistant serovars of Salmonella enterica isolated from foods in Germany. J Antimicrob Chemother. 2005;56(6):1025–33.PubMedCrossRefGoogle Scholar
  15. 15.
    Gomez TM, Motarjemi Y, Miyagawa S, Kaferstein FK, Stohr K. Foodborne salmonellosis. World health statistics quarterly Rapport trimestriel de statistiques sanitaires mondiales. 1997;50(1–2):81–9.PubMedGoogle Scholar
  16. 16.
    Angulo FJ, Nargund VN, Chiller TC. Evidence of an association between use of anti-microbial agents in food animals and anti-microbial resistance among bacteria isolated from humans and the human health consequences of such resistance. J Vet Med B Infect Dis Vet Public Health. 2004;51(8–9):374–9.PubMedCrossRefGoogle Scholar
  17. 17.
    Threlfall EJ. Antimicrobial drug resistance in Salmonella: problems and perspectives in food- and water-borne infections. FEMS Microbiol Rev. 2002;26(2):141–8.PubMedCrossRefGoogle Scholar
  18. 18.
    Brown DJ, Mather H, Browning LM, Coia JE. Investigation of human infections with Salmonella enterica serovar Java in Scotland and possible association with imported poultry. Eur Surveill. 2003;8(2):35–40.Google Scholar
  19. 19.
    Papavasileiou K, Papavasileiou E, Tseleni-Kotsovili A, Bersimis S, Nicolaou C, Ioannidis A, et al. Comparative antimicrobial susceptibility of biofilm versus planktonic forms of Salmonella enterica strains isolated from children with gastroenteritis. Eur J Clin Microbiol Infect Dis. 2010;29(11):1401–5.PubMedCrossRefGoogle Scholar
  20. 20.
    Olson ME, Ceri H, Morck DW, Buret AG, Read RR. Biofilm bacteria: formation and comparative susceptibility to antibiotics. Can J Vet Res. 2002;66(2):86–92.PubMedGoogle Scholar
  21. 21.
    Majtan J, Majtanova L, Xu M, Majtan V. In vitro effect of subinhibitory concentrations of antibiotics on biofilm formation by clinical strains of Salmonella enterica serovar Typhimurium isolated in Slovakia. J Appl Microbiol. 2008;104(5):1294–301.PubMedCrossRefGoogle Scholar
  22. 22.
    Kaplan JB. Antibiotic-induced biofilm formation. Int J Artif Organs. 2011;34(9):737–51.PubMedCrossRefGoogle Scholar
  23. 23.
    Mah TF. Biofilm-specific antibiotic resistance. Future Microbiol. 2012;7(9):1061–72.PubMedCrossRefGoogle Scholar
  24. 24.
    Guiney DG. The role of host cell death in Salmonella infections. Curr Topics Microbiol Immunol. 2005;289:131–50.CrossRefGoogle Scholar
  25. 25.
    Haraga A, Miller SI. A Salmonella enterica serovar typhimurium translocated leucine-rich repeat effector protein inhibits NF-kappa B-dependent gene expression. Infect Immun. 2003;71(7):4052–8.PubMedCrossRefGoogle Scholar
  26. 26.
    Collier-Hyams LS, Zeng H, Sun J, Tomlinson AD, Bao ZQ, Chen H, et al. Cutting edge: Salmonella AvrA effector inhibits the key proinflammatory, anti-apoptotic NF-kappa B pathway. J Immunol. 2002;169(6):2846–50.PubMedGoogle Scholar
  27. 27.
    Raffatellu M, Wilson RP, Chessa D, Andrews-Polymenis H, Tran QT, Lawhon S, et al. SipA, SopA, SopB, SopD, and SopE2 contribute to Salmonella enterica serotype typhimurium invasion of epithelial cells. Infection and immunity. 2005;73(1):146–54.PubMedCrossRefGoogle Scholar
  28. 28.
    Boonyom R, Karavolos MH, Bulmer DM, Khan CM. Salmonella pathogenicity island 1 (SPI-1) type III secretion of SopD involves N- and C-terminal signals and direct binding to the InvC ATPase. Microbiology. 2010;156(Pt 6):1805–14.PubMedCrossRefGoogle Scholar
  29. 29.
    Friebel A, Ilchmann H, Aepfelbacher M, Ehrbar K, Machleidt W, Hardt WD. SopE and SopE2 from Salmonella typhimurium activate different sets of RhoGTPases of the host cell. J Biol Chem. 2001;276(36):34035–40.PubMedCrossRefGoogle Scholar
  30. 30.
    Wang YP, Li L, Shen JZ, Yang FJ, Wu YN. Quinolone-resistance in Salmonella is associated with decreased mRNA expression of virulence genes invA and avrA, growth and intracellular invasion and survival. Vet Microbiol. 2009;133(4):328–34.PubMedCrossRefGoogle Scholar
  31. 31.
    Hur J, Choi YY, Park JH, Jeon BW, Lee HS, Kim AR, et al. Antimicrobial resistance, virulence-associated genes, and pulsed-field gel electrophoresis profiles of Salmonella enterica subsp. enterica serovar Typhimurium isolated from piglets with diarrhea in Korea. Can J Vet Res. 2011;75(1):49–56.PubMedGoogle Scholar
  32. 32.
    Smith KP, George J, Cadle KM, Kumar S, Aragon SJ, Hernandez RL, et al. Elucidation of antimicrobial susceptibility profiles and genotyping of Salmonella enterica isolates from clinical cases of salmonellosis in New Mexico in 2008. World J Microbiol Biotechnol. 2010;26(6):1025–31.PubMedCrossRefGoogle Scholar
  33. 33.
    Hughes LA, Shopland S, Wigley P, Bradon H, Leatherbarrow AH, Williams NJ, et al. Characterisation of Salmonella enterica serotype Typhimurium isolates from wild birds in northern England from 2005–2006. BMC Vet Res. 2008;4:4.PubMedCrossRefGoogle Scholar
  34. 34.
    Prager R, Rabsch W, Streckel W, Voigt W, Tietze E, Tschape H. Molecular properties of Salmonella enterica serotype paratyphi B distinguish between its systemic and its enteric pathovars. J Clin Microbiol. 2003;41(9):4270–8.PubMedCrossRefGoogle Scholar
  35. 35.
    Eberl L, Riedel K. Mining quorum sensing regulated proteins—role of bacterial cell-to-cell communication in global gene regulation as assessed by proteomics. Proteomics. 2011;11(15):3070–85.PubMedCrossRefGoogle Scholar
  36. 36.
    Coenye T. Response of sessile cells to stress: from changes in gene expression to phenotypic adaptation. FEMS Immunol Med Microbiol. 2010;59(3):239–52.PubMedGoogle Scholar
  37. 37.
    Wood MW, Rosqvist R, Mullan PB, Edwards MH, Galyov EE. SopE, a secreted protein of Salmonella dublin, is translocated into the target eukaryotic cell via a sip-dependent mechanism and promotes bacterial entry. Mol Microbiol. 1996;22(2):327–38.PubMedCrossRefGoogle Scholar
  38. 38.
    Graziani C, Busani L, Dionisi AM, Caprioli A, Ivarsson S, Hedenstrom I, et al. Virulotyping of Salmonella enterica serovar Napoli strains isolated in Italy from human and nonhuman sources. Foodborne Pathog Dis. 2011;8(9):997–1003.PubMedCrossRefGoogle Scholar
  39. 39.
    Velge P, Wiedemann A, Rosselin M, Abed N, Boumart Z, Chausse AM, et al. Multiplicity of Salmonella entry mechanisms, a new paradigm for Salmonella pathogenesis. MicrobiologyOpen. 2012;1(3):243–58.PubMedCrossRefGoogle Scholar
  40. 40.
    Boyd EF. Bacteriophage-encoded bacterial virulence factors and phage-pathogenicity island interactions. Adv Virus Res. 2012;82:91–118.PubMedCrossRefGoogle Scholar
  41. 41.
    Srikanth CV, Mercado-Lubo R, Hallstrom K, McCormick BA. Salmonella effector proteins and host-cell responses. Cell Mol Life Sci. 2011;68(22):3687–97.PubMedCrossRefGoogle Scholar
  42. 42.
    Bonizzi G, Karin M. The two NF-kappaB activation pathways and their role in innate and adaptive immunity. Trends Immunol. 2004;25(6):280–8.PubMedCrossRefGoogle Scholar
  43. 43.
    Memet S. NF-kappaB functions in the nervous system: from development to disease. Biochem Pharmacol. 2006;72(9):1180–95.PubMedCrossRefGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2013

Authors and Affiliations

  • A. Ioannidis
    • 1
  • K. Papavasileiou
    • 2
  • E. Papavasileiou
    • 2
  • S. Bersimis
    • 3
  • S. Chatzipanagiotou
    • 2
  1. 1.Department of Nursing, Faculty of Human Movement and Quality of Life SciencesUniversity of PeloponneseSpartaGreece
  2. 2.Department of Medical Biopathology and Clinical Microbiology, Athens Medical School, Aeginition HospitalUniversity of AthensAthensGreece
  3. 3.Department of Statistics and Insurance ScienceUniversity of PiraeusPiraeus Greece

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