Determination the site of antibiotic resistance genes in Escherichia coli isolated From Urinary Tract Infection

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Karzan Abdulmuhsin Mohammad Zirak F. Ahmed Bayar A Mohammed Rasti H Saeed

Abstract

This study includes isolation of 25 isolates of Escherichia coli (E. coli  ) strain from urinary tract samples in a pregnant woman. Microbiological and biochemical tests were used to identify the resistant bacteria of this genus. Screening methods were used to determine bacterial isolates for their resistance to 10 antibiotics include: Amikacin (Ak), Amoxicillin (Ax), Ampicillin (Ap), Chloramphenicol (Cm), Ciprofloxacin (Cip), Erythromycin (Er), Nalidixic acid (Nal), Penicillin (Pen), Tetracycline (Tet) and Trimethoprim (Tm). The isolates E4, E9, E16, and E17 were resistant to all antibiotics used in the current  study using the disk diffusion method. In contrast, the resistance percentage for all antibiotics ranged between 28-96%. Sites of resistance genes and hemolysin production genes were characterized by tranformation techniques in the E4 and E16. The results showed that the antibiotic resistance genes of Amikacin, Erythromycin, Tetracyclin, and Trimethoprim were located on a plasmid, whereas Amoxicillin, Ampicillin, Chloramphenicol, Ciprofloxacin, Nalidixic acid and Penicillin were located on chromosomal DNA. The results also demonstrated an inability to produce alpha or beta-hemolysin indicating that the genes which are responsible for hemolysin production were also located on chromosomal DNA. 

Keywords

E. coli , antibiotics, resistant genes, plasmid

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[1] C. W. Tan and M. P. Chlebicki, "Urinary tract infections in adults,"Singapore Med J, vol. 57, pp. 485-90, Sep 2016.
[2] A. Al-Faisal, Genetic engineering, Dar Al-Shrooq for publishing, Amman, Jordan, 1999.
[3] K.C. Carroll, S.A. Morse, T. Mietzner, and S. Miller, Jawetz, Melnick, and Adelberg’s Medical Microbiology, 27th Ed. McGraw‒Hill Education, USA, 2016.
[4] R. M. Atlas, A. E. Broun and L. C. Parks, Laboratory manual of experimental microbiology, Mosby, St. Louis Baltimore, U.S.A. 1995.
[5] P. Hawky and D. Lewis, Medical bacteriology, 2nd edition, Oxford University Press. Inc, UK, 2004.
[6] H. C. Brinboim and J. Doly, ʺA rapid alkaline extraction procedure for screening recombination plasmid DNA,ʺ, Nucleic acid research. Vol. 7, pp. 1513-1524, 1979.
[7] J. Sambrook and D. W. Russell, Molecular cloning: a laboratory manual, 3rd edition, Cold spring, Herbour Lab., New York. USA, 2001.
[8] M. Mandel and A. Higa, "Calcium-dependent bacteriophage DNA infection. 1970," Biotechnology, vol. 24, pp. 198-201, 1992.
[9] E. M. Lederberg and S. N. Cohen, "Transformation of Salmonella typhimurium by plasmid deoxyribonucleic acid," J Bacteriol, vol. 119, pp. 1072-4, Sep 1974.
[10] W. P. M. Hoekstra, H. E. N. Bergmans, and E. M. Zuidweg, “Transformation in Escherichia coli: studies on the nature of donor DNA after uptake and integration,” Genetical Research, vol. 35, no. 3, pp. 279–289, 1980.
[11] C. Turpin, B. Minkah, K. Danso, and E. Frimpong, "Asymptomatic bacteriuria in pregnant women attending antenatal clinic at komfo anokye teaching hospital, kumasi, ghana," Ghana Med J, vol. 41, pp. 26-9, Mar 2007.
[12] J. R. Johnson, "Virulence factors in Escherichia coli urinary tract infection," Clin Microbiol Rev, vol. 4, pp. 80-128, Jan 1991.
[13] J. E. Blanco, M. Blanco, A. Mora, and J. Blanco, "Prevalence of bacterial resistance to quinolones and other antimicrobials among avian Escherichia coli strains isolated from septicemic and healthy chickens in Spain," J Clin Microbiol, vol. 35, pp. 2184-5, Aug 1997.
[14] S. Sharma, G. K. Bhat, and S. Shenoy, "Virulence factors and drug resistance in Escherichia coli isolated from extraintestinal infections," Indian J Med Microbiol, vol. 25, pp. 369-73, Oct 2007.
[15] G. Wistreich, A. Kleemm and M. D. Lechtman, Laboratory exercises in Microbiology, 4th ed., Glencoe publishing Co. Inc, New York, 1980.
[16] A. E. Barber, B. A. Fleming, and M. A. Mulvey, "Similarly Lethal Strains of Extraintestinal Pathogenic Escherichia coli Trigger Markedly Diverse Host Responses in a Zebrafish Model of Sepsis," mSphere, vol. 1, Mar-Apr 2016.
[17] M. D. Engstrom and H. L. Mobley, "Regulation of Expression of Uropathogenic Escherichia coli Nonfimbrial Adhesin TosA by PapB Homolog TosR in Conjunction with H-NS and Lrp," Infect Immun, vol. 84, pp. 811-21, Jan 11 2016.
[18] M. Abd-Alsattar, Partial purification of type-1pili extracted from E. coli and its role in infection, .Ph.D.Thesis, College of Science, University of Baghdad, 2004.
[19] U. Asad and S. Zaman, ʺMultiple drug resistance pattern in urinary tract infection patients in Aligarh,ʺ Biochemical research, vol. 17, pp.179-181, 2006.
[20] M. H. Shirazi, N. Sadeghhifard, R. Ranjbar, E. Daneshyar and A. Ghasemi, ʺIncidence of asymptomatic bacteriuria during pregnancy, ʺ Pak. J. Biol. Sci. , vol. 9, pp. 151-154, 2006.
[21] A. M. G. Al-Fahdawi, Influence of blood groups on the availability of receptors of uroepithelial cells for attachment of uropathogenic bacteria causing urinary tract infections, M.Sc. thesis, College of Medicine, University of Baghdad, Baghdad, Iraq, 2001
[22] A. N. H. Al-Alosi, Molecular study on some virulence factors produced by Gram-negative bacteria, M.Sc. thesis, Genetic Enginnering and biotechnology Institute for post graduate studies, University of Baghdad, Baghdad, Iraq, 2004.
[23] A. N. Ghosh, D. R. Bhatta, M. T. Ansari, H. K. Tiwari, J. P. Mathuria, A. Gaur, et al., "Application of WHONET in the Antimicrobial Resistance Surveillance of Uropathogens: A First User Experience from Nepal," J Clin Diagn Res, vol. 7, pp. 845-8, May 2013.
[24] M. M. Al-Shaikhli, Virulence factors of E.coli in women with asymptomatic bacteriuria, M.Sc thesis, College of Medicine, Baghdad University, Baghdad. Iraq, 2004.
[25] A. Sharma and P. Grover, ʺApplication of whonet for the surveiallance of antimicrobial resistance, ʺ Indian. J. Med. Microbiol., vol. 22, pp.115-118, 2004.
[26] S. Babypadmini and B. Appalaraju, "Extended spectrum -lactamases in urinary isolates of Escherichia coli and Klebsiella pneumoniae - prevalence and susceptibility pattern in a tertiary care hospital," Indian J Med Microbiol, vol. 22, pp. 172-4, Jul-Sep 2004.
[27] A. Moghadas and G. Irajian, "Asymptomatic urinary tract infection in women," Iranian Journal of Pathology, vol. 4, pp.105-108, Jul 2009.
[28] N. Woodford and A. Johnson, Molecular Bacteriology, Protocols and clinical applications, Humana press Inc, 1998.
[29] J. R. Brown, D. Gentry, J. A. Becker, K. Ingraham, D. J. Holmes, and M. J. Stanhope, "Horizontal transfer of drug-resistant aminoacyl-transfer-RNA synthetases of anthrax and Gram-positive pathogens," EMBO Rep, vol. 4, pp. 692-8, Jul 2003.
[30] P. Nwanze, L. Nwaru, S. Oranusi, U. Dimkpa, M. Okwu, B. Babatunde, T. Anake, W. Jatto and C. Asagwara, ʺUrinary tract infection in Okada village: Prevalence and antimicrobial susceptibility pattern,ʺ Scientific Research and Essay, vol. 2, pp. 112-116 . 2007
[31] Z. A. Nanakaly, Characterization of plasmid DNA content and antimicrobial effect of bile salt on E.coli isolated from urinary tract infection of pregnant women, Thesis of Higher Diploma, College of Education, Salahaddin University, Erbil, Iraq, 2010.
[32] P. Siskova, L. Cernohorska, M. Mahelova, K. Turkova, and V. Woznicova, "Phenotypes of Escherichia coli isolated from urine: Differences between extended-spectrum beta-lactamase producers and sensitive strains," J Microbiol Immunol Infect, vol. 48, pp. 329-34, Jun 2015.