Urine influences growth and virulence gene expressions in Uropathogenic E. coli: a comparison with nutrient limited medium

Year 2021, Volume: 11 Issue: 2, 209 - 214, 30.06.2021

Defne Gümüş Fatma Kalaycı Yüksek Gülşen Uz Özlem Sefer Emre Yörük Mine Küçüker

https://doi.org/10.33808/clinexphealthsci.686302

Abstract

Objective. The interactions between environmental factors and microbial biological process are well known. Urine has been selected to provide host conditions probably affecting bacterial growth and gene expressions. The aim of this study was to detect the modulations of growth and gene expressions [sfa/foc, cnf1, usp and aer] of UPEC in urine by comparing the results with serum supplemented Standart American Petroleum Institute medium which is defined as host-like medium.

Methods. UPECs were incubated 37°C and growth alterations were detected by measuring the changes in the absorbance at 600 nm in four-,six- and 24 hours periods. Gene expression levels were analysed by quantitative polymerase chain reaction (qPCR). Statistical analysis of fold changes in gene expression values and growths were calculated using one-way ANOVA Tukey's post hoc test.

Results. The increase of bacterial growth in urine was found to be statistically significant (p<0.0001). The alterations of aer and sfa/foc expression levels were statistically significant (p<0.001); whereas the expression levels of cnf1 and usp genes were not altered (p>0.05).

Conclusion. According to our results, urine as an environment in vivo affected both the growth and gene expression in UPEC.

Keywords

urine, virulence, growth, gene expression

References

• 1. Freestone P. Communication between bacteria and their hosts. Scientifica 2013. 2. Bakholdina SI, Sanina NM, Krasikova IN, Popova OB & Solov’eva TF. The impact of abiotic factors (temperature and glucose) on physicochemical properties of lipids from Yersinia pseudotuberculosis. Biochimie 2004; 86 (12):875-881. 3. Lyte M. Bacteria and hormones: Why the science of microbial endocrinology matters to disease and well-being. Heidt PJ, Midtvedt T, Volker R & Versalovic J, editors. Bacterial species as partners and pathogens. Germany: Herborn-Dill: Old Herborn University Foundation; 2012. p. 17-29. 4. Hughes DT & Sperandio V. Inter-kingdom signalling: communication between bacteria and their hosts. Nat Rev Microbiol 2008; 6(2): 111-120. 5. Lyte M, Freestone PP. (eds). Microbial Endocrinology: Interkingdom Signaling in Infectious Disease and Health. New York: Springer; 2010. 6. Kendall MM & Sperandio V. What a dinner party! Mechanisms and functions of interkingdom signaling in host-pathogen associations. mBio 2016; 7(2): e01748-15. 7. Hamner S, McInnerney K, Williamson K, Franklin MJ, Ford TE. Bile salts affect expression o Escherichia coli O157: H7 genes for virulence and iron acquisition, and promote growth under iron limiting conditions. PLoS One 2013;8(9):e74647. 8. Hirakawa H, Inazumi Y, Masaki T, Hirata T, Yamaguchi A. Indole induces the expression of multidrug exporter genes in Escherichia coli. Mol Microbiol 2005;55(4):1113-26. 9. Klosowska K & Plotkin BJ. Human insulin modulation of Escherichia coli adherence and chemotaxis. Am J Infect Dis 2006; 2(4): 197-200. 10. Kurakado S, Kurogane R & Sugita T. 17β-Estradiol inhibits estrogen binding protein-mediated hypha formation in Candida albicans. Microb Pathog 2017;109: 151-155. 11. Delcenserie V, LaPointe G, Charaslertrangsi T, Rabalski A, Griffiths MW. Glucose decreases virulence gene expression of Escherichia coli O157: H7. J Food Prot 2012; 75(4):748-52. 12. Gönüllü N, Küçükbasmacı Ö, Büyükbaba-Boral Ö, Anğ-Küçüker M. The influence of glucose added urine on the in vitro antimicrobial activity of various antibiotics. Indian J Med Res 2008; 128(5):663-5. 13. Stamey TA & Mihara G. (1980). Observations on the growth of urethral and vaginal bacteria in sterile urine. J Urol 1980; 124(4): 461-463. 14. Hull RA & Hull SI. Nutritional requirements for growth of uropathogenic Escherichia coli in human urine. Infect Immun 1997; 65(5): 1960-1961. 15. Gordon DM & Riley MA. A theoretical and experimental analysis of bacterial growth in the bladder. Mol Microbiol 1992;6(4): 555-562. 16. Vejborg RM, de Evgrafov MR, Phan MD, Totsika M, Schembri MA & Hancock V. Identification of genes important for growth of asymptomatic bacteriuria Escherichia coli in urine. Infect Immun 2012; 80(9): 3179-3188. 17. Mann R, Mediati DG, Duggin IG, Harry EJ & Bottomley AL. Metabolic adaptations of uropathogenic E. coli in the urinary tract. Front Cell Infect Microbiol 2017; 7: 241. 18. Greene SE, Hibbing ME, Janetka J, Chen SL & Hultgren SJ. Human urine decreases function and expression of type 1 pili in uropathogenic Escherichia coli. mBio, 2015; 6(4): e00820-15. 19. Hancock V & Klemm P. Global gene expression profiling of asymptomatic bacteriuria Escherichia coli during biofilm growth in human urine. Infect Immun 2007; 75(2): 966-976. 20. Schwan WR, Lee JL, Lenard FA, Matthews BT & Beck MT. Osmolarity and pH growth conditions regulate fim gene transcription and type 1 pilus expression in uropathogenic Escherichia coli. Infect Immun 2002;70(3):1391-1402. 21. Thornton LA, Burchell RK, Burton SE, Lopez‐Villalobos N, Pereira D, MacEwan I, ... & Velathanthiri N. (2018). The effect of urine concentration and pH on the growth of Escherichia coli in canine urine in vitro. J Vet Intern Med 2018;32(2): 752-756. 22. Russo TA, Carlino UB, Mong A & Jodush ST. (1999). Identification of genes in an extraintestinal isolate of Escherichia coli with increased expression after exposure to human urine. Infect Immun 1999; 67(10): 5306-5314. 23. Freestone PP, Haigh RD, Williams PH & Lyte M. Stimulation of bacterial growth by heat-stable, norepinephrine-induced autoinducers. FEMS Microbiol Lett 1999;172(1): 53-60. 24. Freestone PP, Sandrini SM, Haigh RD & Lyte M. Microbial endocrinology: how stress influences susceptibility to infection. Trends Microbiol 2008;16(2): 55-64. 25. Andersen-Civil AIS, Ahmed S, Guerra PR, Andersen TE, Hounmanou YMG, Olsen JE & Herrero-Fresno A. The impact of inactivation of the purine biosynthesis genes, purN and purT, on growth and virulence in uropathogenic E. coli. Mol Biol Rep 2018; 45(6): 2707-2716. 26. King JE, Owaif HAA, Jia J & Roberts IS. Phenotypic heterogeneity in expression of the K1 polysaccharide capsule of uropathogenic Escherichia coli and downregulation of the capsule genes during growth in urine. Infect Immun 2015; 83(7): 2605-2613. 27. Schwan WR, Beck MT, Hung CS & Hultgren SJ. Differential regulation of Escherichia coli fim genes following binding to mannose receptors. J pathog 2018. 28. Altman PL. Physical properties and chemical composition of urine: mammals. Dittmer DL, editor. Part 1 : Man. In Blood and Other Body Fluids. Washington, DC : Federation of American Societies for Experimental Biology; 1961. p. 363–369. 29. Asscher AW, Sussman M & Weisser R. Bacterial growth in human urine. O’Grady F & Brumfitt W, editors. In Urinary Tract Infection. London : Oxford University Press; 1968. p. 3–13. 30. Asscher AW, Sussman M, Waters WE, Davis RH & Chick S. Urine as a medium for bacterial growth. Lancet 1966; 2: 1037. 31. Gumus D, Yoruk E, Kalayci-Yuksek F, Uz G, Topal-Sarikaya A, Ang-Kucuker M. The effects of insulin and glucose on different characteristics of a UPEC: Alterations in growth rate and expression levels of some virulence genes. Clin Lab 2017; 63(10): 1589-97. 32. Gümüş D, Kalaycı-Yüksek F, Uz G, Bilgin M, Anğ-Küçüker M. The Possible Effects of Different Hormones on Growth Rate and Ability of Biofilm Formation in Different Types of Microorganisms. Acta Microbiol Bulg 2018; 34(1): 47-51. 33. Gümüş D, Yüksek FK, Sefer Ö, Yörük E, Uz G, Küçüker MA. The roles of hormones in the modulation of growth and virulence genes’ expressions in UPEC strains. Microb Pathog 2019; 132:319-324. 34. Roos V, Ulett GC, Schembri MA & Klemm P. The asymptomatic bacteriuria Escherichia coli strain 83972 out competes uropathogenic E. coli strains in human urine. Infect Immun 2006; 74(1): 615-624.