Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지
DOI QR코드

DOI QR Code

Role of eptC in Biofilm Formation by Campylobacter jejuni NCTC11168 on Polystyrene and Glass Surfaces

  • Lim, Eun Seob (Department of Food Biotechnology, Korea University of Science and Technology) ;
  • Kim, Joo-Sung (Department of Food Biotechnology, Korea University of Science and Technology)
  • Received : 2016.10.18
  • Accepted : 2017.07.05
  • Published : 2017.09.28
Download PDF
⟨ Previous Next ⟩

Abstract

The complex roles of cell surface modification in the biofilm formation of Campylobacter jejuni, a major cause of worldwide foodborne diarrheal disease, are poorly understood. In a screen of mutants from random transposon mutagenesis, an insertional mutation in the eptC gene (cj0256) resulted in a significant decrease in C. jejuni NCTC11168 biofilm formation (<20%) on major food contact surfaces, such as polystyrene and borosilicate glass, when compared with wild-type cells (p < 0.05). In C. jejuni strain 81-176, the protein encoded by eptC modified cell surface structures, such as lipid A, the inner core of lipooligosaccharide, and the flagellar rod protein (FlgG), by attaching phosphoethanolamine. To assess the role of eptC in C. jejuni NCTC11168, adherence and motility tests were performed. In adhesion assays with glass surfaces, the eptC mutant exhibited a $0.77log\;CFU/cm^2$ decrease in adherence compared with wild-type cells during the initial 2 h of the assay (p < 0.05). These results support the hypothesis that the modification of cell surface structures by eptC affects the initial adherence in biofilm formation of C. jejuni NCTC11168. In motility tests, the eptC mutant demonstrated reduced motility when compared with wild-type cells, but wild-type cells with the transposon inserted in a gene irrelevant to biofilm formation (cj1111c) also exhibited decreased motility to a similar extent as the eptC mutant. This suggests that although eptC affects motility, it does not significantly affect biofilm formation. This study demonstrates that eptC is essential for initial adherence, and plays a significant role in the biofilm formation of C. jejuni NCTC11168.

Keywords

References

  1. Luber P, Brynestad S, Topsch D, Scherer K, Bartelt E. 2006. Quantification of Campylobacter species cross-contamination during handling of contaminated fresh chicken parts in kitchens. Appl. Environ. Microbiol. 72: 66-70. https://doi.org/10.1128/AEM.72.1.66-70.2006
  2. Olson CK, Ethelberg S, Pelt W van, Tauxe RV. 2008. Epidemiology of Campylobacter jejuni infections in industrialized nations. In Nachamkin I, Szymanski CM, Blaser MJ (eds), Campylobacter, 3rd Ed. American Society of Microbiology, Washington, DC.
  3. MFDS. Ministry of Food and Drug Safety. 2015. Occurrence of foodborne disease from 2003-2014. Available from http://www. foodsafetykorea.go.kr/portal/healthyfoodlife/foodPoisoningStat. do?menu_no=5 19&menu_grp=MENU_GRP02.
  4. Costerton J W, S tewart P S, G reenberg EP. 1 999. Bacterial biofilms: a common cause of persistent infections. Science 284: 1318-1322. https://doi.org/10.1126/science.284.5418.1318
  5. Bridier A, Sanchez-Vizuete P, Guilbaud M, Piard JC, Naïtali M, Briandet R. 2015. Biofilm-associated persistence of food-borne pathogens. Food Microbiol. 45: 167-178. https://doi.org/10.1016/j.fm.2014.04.015
  6. Gunther NW, Chen CY. 2009. The biofilm forming potential of bacterial species in the genus Campylobacter. Food Microbiol. 26: 44-51. https://doi.org/10.1016/j.fm.2008.07.012
  7. Joshua GWP, Guthrie-Irons C, Karlyshev AV, Wren BW. 2006. Biofilm formation in Campylobacter jejuni. Microbiology 152: 387-396. https://doi.org/10.1099/mic.0.28358-0
  8. Kalmokoff M, Lanthier P, Tremblay TL, Foss M, Lau PC, Sanders G, et al. 2006. Proteomic analysis of Campylobacter jejuni 11168 biofilms reveals a role for the motility complex in biofilm formation. J. Bacteriol. 188: 4312-4320. https://doi.org/10.1128/JB.01975-05
  9. Kim JS, Park CW, Kim YJ. 2015. Role of flgA for flagellar biosynthesis and biofilm formation of Campylobacter jejuni NCTC11168. J. Microbiol. Biotechnol. 25: 1871-1879. https://doi.org/10.4014/jmb.1504.04080
  10. Fields JA, Thompson SA. 2008. Campylobacter jejuni CsrA mediates oxidative stress responses, biofilm formation, and host cell invasion. J. Bacteriol. 190: 3411-3416. https://doi.org/10.1128/JB.01928-07
  11. Bronowski C, James CE, Winstanley C. 2014. Role of environmental survival in transmission of Campylobacter jejuni. FEMS Microbiol. Lett. 356: 8-19. https://doi.org/10.1111/1574-6968.12488
  12. Lin J, Wang Y, Hoang K Van. 2009. Systematic identification of genetic loci required for polymyxin resistance in Campylobacter jejuni using an efficient in vivo transposon mutagenesis system. Foodborne Pathog. Dis. 6: 173-185. https://doi.org/10.1089/fpd.2008.0177
  13. Cullen TW, Trent MS. 2010. A link between the assembly of flagella and lipooligosaccharide of the gram-negative bacterium Campylobacter jejuni. Proc. Natl. Acad. Sci. USA 107: 5160-5165. https://doi.org/10.1073/pnas.0913451107
  14. Watnick P, Kolter R. 2000. Biofilm, city of microbes. J. Bacteriol. 182: 2675-2679. https://doi.org/10.1128/JB.182.10.2675-2679.2000
  15. Abee T, Kovacs AT, Kuipers OP, Veen SVD. 2011. Biofilm formation and dispersal in gram-positive bacteria. Curr. Opin. Biotechnol. 22: 172-179. https://doi.org/10.1016/j.copbio.2010.10.016
  16. Stoodley P, Sauer K, Davies DG, Costerton JW. 2002. Biofilms as complex differentiated communities. Annu. Rev. Microbiol. 56: 187-209. https://doi.org/10.1146/annurev.micro.56.012302.160705
  17. Scott NE, Nothaft H, Edwards AVG, Labbate M, Djordjevic SP, Larsen MR, et al. 2012. Modification of the Campylobacter jejuni N-linked glycan by EptC protein-mediated addition of phosphoethanolamine. J. Biol. Chem. 287: 29384-29396. https://doi.org/10.1074/jbc.M112.380212
  18. Cullen TW, O'Brien JP, Hendrixson DR, Giles DK, Hobb RI, Thompson SA, et al. 2013. EptC of Campylobacter jejuni mediates phenotypes involved in host interactions and virulence. Infect. Immun. 81: 430-440. https://doi.org/10.1128/IAI.01046-12
  19. Cullen TW, Madsen JA, Ivanov PL, Brodbelt JS, Trent MS. 2012. Characterization of unique modification of flagellar rod protein FlgG by Campylobacter jejuni lipid A phosphoethanolamine transferase, linking bacterial locomotion and antimicrobial peptide resistance. J. Biol. Chem. 287: 3326-3336. https://doi.org/10.1074/jbc.M111.321737
  20. Donlan RM. 2002. Biofilms: microbial life on surfaces. Emerg. Infect. Dis. 8: 881-890. https://doi.org/10.3201/eid0809.020063
  21. O'Toole GA, Kolter R. 1998. Initiation of biofilm formation in Pseudomonas fluorescens WCS365 proceeds via multiple, convergent signalling pathways: a genetic analysis. Mol. Microbiol. 28: 449-461. https://doi.org/10.1046/j.1365-2958.1998.00797.x
  22. O'Toole GA, Kolter R. 1998. Flagellar and twitching motility are necessary for Pseudomonas aeruginosa biofilm development. Mol. Microbiol. 30: 295-304. https://doi.org/10.1046/j.1365-2958.1998.01062.x
  23. Svensson SL, Pryjma M, Gaynor EC. 2014. Flagella-mediated adhesion and extracellular DNA release contribute to biofilm formation and stress tolerance of Campylobacter jejuni. PLoS One 9: e106063. https://doi.org/10.1371/journal.pone.0106063
  24. Moe KK, Mimura J, Ohnishi T, Wake T, Yamazaki W, Nakai M, et al. 2010. The mode of biofilm formation on smooth surfaces by Campylobacter jejuni. J. Vet. Med. Sci. 72: 411-416. https://doi.org/10.1292/jvms.09-0339
  25. Reeser RJ, Medler RT, Billington SJ, Jost BH, Joens LA. 2007. Characterization of Campylobacter jejuni biofilms under defined growth conditions. Appl. Environ. Microbiol. 73: 1908-1913. https://doi.org/10.1128/AEM.00740-06
  26. Golden NJ, Acheson DWK. 2002. Identification of motility and autoagglutination Campylobacter jejuni mutants by random transposon mutagenesis. Infect. Immun. 70: 1761-1771. https://doi.org/10.1128/IAI.70.4.1761-1771.2002
  27. Grant CCR, Konkel ME, Cieplak WJ, Tompkins LS. 1993. Role of flagella in adherence, internalization, and translocation of Campylobacter jejuni in nonpolarized and polarized epithelial cell cultures. Infect. Immun. 61: 1764-1771.
  28. Young NM, B risson J R, K elly J , Watson D C, T essier L , Lanthier PH, et al. 2002. Structure of the N-linked glycan present on multiple glycoproteins in the gram-negative bacterium, Campylobacter jejuni. J. Biol. Chem. 277: 42530-42539. https://doi.org/10.1074/jbc.M206114200
  29. Nguyen VT, Turner MS, Dykes GA. 2011. Influence of cell surface hydrophobicity on attachment of Campylobacter to abiotic surfaces. Food Microbiol. 28: 942-950. https://doi.org/10.1016/j.fm.2011.01.004
  30. Martino PD, Cafferini N, Joly B, Darfeuille-Michaud A. 2003. Klebsiella pneumoniae type 3 pili facilitate adherence and biofilm formation on abiotic surfaces. Res. Microbiol. 154: 9-16. https://doi.org/10.1016/S0923-2508(02)00004-9
  31. Moser I, Schröder W. 1997. Hydrophobic characterization of thermophilic Campylobacter species and adhesion to INT 407 cell membranes and fibronectin. Microb. Pathog. 22: 155-164. https://doi.org/10.1006/mpat.1996.0104
  32. Raetz CRH, Reynolds CM, Trent MS, Bishop RE. 2007. Lipid A modification systems in gram-negative bacteria. Annu. Rev. Biochem. 76: 295-329. https://doi.org/10.1146/annurev.biochem.76.010307.145803
  33. Kannenberg EL, Carlson RW. 2001. Lipid A and O-chain modifications cause Rhizobium lipopolysaccharides to become hydrophobic during bacteroid development. Mol. Microbiol. 39: 379-391. https://doi.org/10.1046/j.1365-2958.2001.02225.x
  34. Hankins JV, Madsen JA, Giles DK, Brodbelt JS, Trent MS. 2012. Amino acid addition to Vibrio cholerae LPS establishes a link between surface remodeling in gram-positive and gramnegative bacteria. Proc. Natl. Acad. Sci. USA 109: 8722-8727. https://doi.org/10.1073/pnas.1201313109
  35. Alemka A, Nothaft H, Zheng J, Szymanski CM. 2013. Nglycosylation of Campylobacter jejuni surface proteins promotes bacterial fitness. Infect. Immun. 81: 1674-1682. https://doi.org/10.1128/IAI.01370-12
  36. Karlyshev AV, Everest P, Linton D, Cawthraw S, Newell DG, Wren BW. 2004. The Campylobacter jejuni general glycosylation system is important for attachment to human epithelial cells and in the colonization of chicks. Microbiology. 150: 1957-1964. https://doi.org/10.1099/mic.0.26721-0

Cited by

  1. Bio-enzymes for inhibition and elimination of Escherichia coli O157:H7 biofilm and their synergistic effect with sodium hypochlorite vol.9, pp.None, 2017, https://doi.org/10.1038/s41598-019-46363-w
  2. Inhibition of Microbial Quorum Sensing Mediated Virulence Factors by Pestalotiopsis sydowiana vol.30, pp.4, 2017, https://doi.org/10.4014/jmb.1907.07030
  3. Campylobacter Biofilms: Potential of Natural Compounds to Disrupt Campylobacter jejuni Transmission vol.22, pp.22, 2017, https://doi.org/10.3390/ijms222212159
  4. A novel high-content screening approach for the elucidation of C. jejuni biofilm composition and integrity vol.21, pp.1, 2021, https://doi.org/10.1186/s12866-020-02062-5
  5. Antimicrobial resistance and genomic diversity of Campylobacter jejuni isolates from broiler caeca and neck skin samples collected at key stages during processing. vol.135, pp.None, 2017, https://doi.org/10.1016/j.foodcont.2021.108664