Iranian Journal of Microbiology 2017. 9(2):89-96.

Role of antigen-43 on biofilm formation and horizontal antibiotic resistance gene transfer in non-O157 Shiga toxin producing Escherichia coli strains
Roholla Taghadosi, Mohammad Reza Shakibaie, Reza Ghanbarpour, Hossein Hosseini-Nave

Abstract


Background and Objectives: The objectives of this study were to evaluate the antibiotic resistance profiles, biofilm formation, presence of antigen 43 (Ag43) gene, and transfer of antibiotic resistance phenotype among non-O157 Shiga toxin producing Escherichia coli (STEC).
Materials and Methods: From October 2014 to November 2015 a total of 276 stool samples were collected from healthy calves, goats and 395 patients with the sign of nonbloody diarrhea and screened for presence of stx and serotype O157 genes by polymerase chain reaction (PCR) technique. Susceptibility to 14 antibiotics was determined as per CLSI guideline. Presence of Ag43 and intimin (eaeA) genes were detected by PCR. Biofilm formation was measured by microtiter plate method. Conjugation was carried out by membrane filter technique.
Results: We isolated 74 (93.6%) non-O157 STEC strains from 41 calves, 33 goats and 5 (6.3%) patients’ stools, however, no O157 serotype was detected in our study. Resistance was observed most commonly to tobramycin (66.2%), kanamycin (48.6%), and amikacin (29.7%) and less frequently to ciprofloxacin (4.1%), amoxicillin-clavulanic acid (5.4%), and ceftriaxone (9.5%) in isolates recovered from calves and goats fecal samples, whereas, all human isolates were sensitive to ceftazidime, ciprofloxacin, tobramycin and imipenem, respectively. Furthermore, Ag43 was detected in 60 STEC isolated from animals and 5 human origins (no eaeA gene was found in this study). Biofilm formation from Ag43+ and Ag43- colonies showed 20 isolates with strong biofilm activities. Cefotaxime resistance phenotype was transferred to E. coli ATCC 25922.1 (Nalr) by conjugation at a frequency of 1.6×10-4.
Conclusion: From the above results we concluded that, human infections with non-O157 STEC were significantly low in Kerman. Ag43 was insignificant with biofilm quantity in most cases.


Keywords


Shiga toxin producing E. coli, Antibiotic resistance, Biofilm, PCR, Conjugation

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References


Hall-Stoodley L, Costerton JW, Stoodley P. Bacterial biofilms: from the natural environment to infectious diseases. Nat Rev Microbiol 2004;2:95-108.

Parsek MR, Singh PK. Bacterial biofilms: an emerging link to disease pathogenesis. Annu Rev Microbiol 2003;57:677-701.

Watnick PI, Kolter R. Steps in the development of a Vibrio cholerae El Tor biofilm. Mol Microbiol 1999;34:586-595.

Danese PN, Pratt LA, Kolter R. Exopolysaccharide production is required for development of Escherichia coli K-12 biofilm architecture. J Bacteriol 2000;182:3593-3596.

Taghadosi R, Shakibaie MR, Masoumi S. Biochemical detection of N-Acyl homoserine lactone from biofilm-forming uropathogenic Escherichia coli isolated from urinary tract infection samples. Reports Biochem Mol Biol 2015;3:56-72.

Ferdous M, Friedrich AW, Grundmann H, de Boer RF, Croughs PD, Islam MA, et al. Molecular characterization and phylogeny of Shiga toxin-producing E. coli (STEC) isolates obtained from two Dutch regions using whole genome sequencing. Clin Microbiol Infect 2016;22:1-9.

Scallan E, Hoekstra RM, Mahon BE, Jones TF, Griffin PM. An assessment of the human health impact of seven leading foodborne pathogens in the United States using disability adjusted life years. Epidemiol Infect 2015;143:2795-2804.

Hussein HS, Bollinger LM. Prevalence of Shiga toxin–producing Escherichia coli in beef cattle. J Food Prot 2005;68:2224-2241.

Tozzi AE, Caprioli A, Minelli F, Gianviti A, De Petris L, Edefonti A, et al. Shiga Toxin–producing Escherichia coli infections associated with hemolytic uremic syndrome. Emerg Infect Dis 2003;9:106-108.

Biscola FT, Abe CM, Guth BEC. Determination of adhesin gene sequences in, and biofilm formation by, O157 and non-O157 Shiga toxin-producing Escherichia coli strains isolated from different sources. Appl Environ Microbiol 2011;77:2201-2208.

Wang R, Bono JL, Kalchayanand N, Shackelford S, Harhay DM. Biofilm formation by Shiga toxin–producing Escherichia coli O157: H7 and non-O157 strains and their tolerance to sanitizers commonly used in the food processing environment. J Food Prot 2012;75:1418-1428.

Picozzi C, Antoniani D, Vigentini I, Foschino R, Kneifel W. Genotypic characterization and biofilm formation of Shiga-toxin-producing Escherichia coli. FEMS Microbiol Lett 2017;364(2). doi: 10.1093/femsle/fnw291.

Schembri MA, Hjerrild L, Gjermansen M, Klemm P. Differential expression of the Escherichia coli autoaggregation factor antigen 43. J Bacteriol 2003;185:2236-2242.

Hasman H, Chakraborty T, Klemm P. Antigen-43-mediated autoaggregation of Escherichia coli is blocked by fimbriation. J Bacteriol 1999;181:4834-4841.

Kjærgaard K, Schembri MA, Ramos C, Molin S, Klemm P. Antigen 43 facilitates formation of multispecies biofilms. Environ Microbiol 2000;2:695-702.

Danese PN, Pratt LA, Dove SL, Kolter R. The outer membrane protein, Antigen 43, mediates cell to cell interactions within Escherichia coli biofilms. Mol Microbiol 2000;37:424-432.

Pratt LA, Kolter R. Genetic analysis of Escherichia coli biofilm formation: roles of flagella, motility, chemotaxis and type I pili. Mol Microbiol 1998;30:285-293.

O’Toole GA, Kolter R. Flagellar and twitching motility are necessary for Pseudomonas aeruginosa biofilm development. Mol Microbiol 1998;30:295-304.

Schroeder CM, Meng J, Zhao S, Debroy C, Torcolini J, Zhao C, et al. Antimicrobial resistance of Escherichia coli O26, O103, O111, O128, and O145 from animals and humans. Emerg Infect Dis 2002;8:1409-1414.

Clinical and Laboratory Standards Institute (CLSI); 2010. Perform stand antimicrob disk susceptibility Tests, Wayne PA.

Mendez-Arancibia E, Vargas M, Soto S, Ruiz J, Kahigwa E, Schellenberg D, et al. Prevalence of different virulence factors and biofilm production in enteroaggregative Escherichia coli isolates causing diarrhea in children in Ifakara (Tanzania). Am J Trop Med Hyg 2008;78:985-989.

Shakibaie MR, Dhakephalker PK, Kapadnis BP, Salajaghe GA, Chopade BA. Plasmid mediated silver and antibiotic resistance in Acinetobacter baumannii BL54. Iran J Med Sci 1998;23:30-36.

Feliciello I, Chinali G. A modified alkaline lysis method for the preparation of highly purified plasmid DNA from Escherichia coli. Anal Biochem 1993;212:394-401.

Lemon KP, Higgins DE, Kolter R. Flagellar motility is critical for Listeria monocytogenes biofilm formation. J Bacteriol 2007;189:4418-4424.

McClaine JW, Ford RM. Characterizing the adhesion of motile and nonmotile Escherichia coli to a glass surface using a parallel plate flow chamber. Biotechnol Bioeng 2002;78:179-189.

Yamamoto T, Fujita K, Yokota T. Adherence characteristics to human small intestinal mucosa of Escherichia coli isolated from patients with diarrhea or urinary tract infections. J Infect Dis 1990;162:896-908.

Girón JA, Torres AG, Freer E, Kaper JB. The flagella of enteropathogenic Escherichia coli mediate adherence to epithelial cells. Mol Microbiol 2002;44:361-379.

Klemm P, Hjerrild L, Gjermansen M, Schembri MA. Structure-function analysis of the self-recognizing antigen 43 autotransporter protein from Escherichia coli. Mol Microbiol 2004;51:283-296.

Ulett GC, Valle J, Beloin C, Sherlock O, Ghigo J-M, Schembri MA. Functional analysis of antigen 43 in uropathogenic Escherichia coli reveals a role in long-term persistence in the urinary tract. Infect Immun 2007;75:3233-3244.

Reisner A, Haagensen JAJ, Schembri MA, Zechner EL, Molin S. Development and maturation of Escherichia coli K-12 biofilms. Mol Microbiol 2003;48:933-946.

Cheney TEA, Smith RP, Hutchinson JP, Brunton LA, Pritchard G, Teale CJ. Cross-sectional survey of antibiotic resistance in Escherichia coli isolated from diseased farm livestock in England and Wales. Epidemiol Infect 2015;143:2653-2659.


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