Analysis of genetic diversity of E.coli bacteria isolates from UTI after exposure to some biological effects by using RAPD technique
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Abstract
Background and Objective: Escherichia coli bacteria are one of the most important bacterial races causing urinary tract infection, which is responsible for 90% of the causes of urinary tract infection and originated in the human intestine, this is Gram- Negative bacteria and facultative aerobic that moves by peripheral flagella and belongs to the bacteria of the Enterobacteriaceae.
Materials and methods: The bacteria were isolated from UTI, after they were diagnosed using light microscope and conducting biochemical tests, then exposed to the antibiotic Ciprofloxacin, trimethoprim, and alcoholic extract of Punica granatum and Trigonella foenum graicum. genomic DNA was extracted for all samples and Random amplified polymorphic DNA - Polymerase chain reaction (RAPD-PCR) marker was carried out using five random primers.
Results: The results of RAPD-PCR profiles shown that exposed to antibiotics (Ciprofloxacin and Trimethoprim) and alcoholic extracts (Punica granatum and Trigonella foenum graicum) lead to the disappearance or appearance new bands compared with non-exposed samples, and the highest rate of polymorphism for all each treatment and primers in sample 5 was 102.27% where the ratio % GTS for all treatment and primers is 6.04% in the same sample.
Conclusion: All treatments caused genetic changes in the DNA of E.coli bacteria cells especially the Punica granatum which gave the highest effect than the rest of the treatments, this indicates its efficiency in treating bacterial infections.
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References
[1] Foxman, B. (2014). Urinary Tract Infection Syndromes Occurrence, Recurrence, Bacteriology, Risk Factors, and Disease Burden. Infect Dis Clin N Am. 28: 1–13.
[2] Forsyth, V. S. Armbruster, C. E.; Smith, S. N.; Pirani, A.; Springman, A.; Walters, M. S.;
Nielubowicz, G. R.; Himpsl, S. D.; Snitkin, E. S.; Mobley, H. L. T. (2018). Rapid Growth of Uropathogenic Escherichia coli During Human Urinary Tract Infection. M bio. 9(2): 1-13.
[3] Brown, A. E., Smith, H. A. (2017). Benson ́ s. Microbiological applications: laboratory manual in general microbiology .14th ed. McGraw-Hill. New York 438 pp.
[4] Weekes, L. M.( 2015). Antibiotic resistance changing management of urinary tract infection in aged care. Med J Aust.; 203(9): 352.
[5] Singh, B.; Tilak, R; Srivastava, R; Katiyar, D. (2014). Urinary Tract infection and its risk factors in Women: an Appraisal. J Pure Appl Microbiol. 2014; 8(5):4155-62.
[6] Wafaa, S. A.W., and Bashair, S.H., (2015). Anti- Virulence Effect of Pomegranate Peel Extracts on Most common urinary tract infection pathogens in pregnant women,Journal of Contem poray Medical
Science. College of science and technology. (1) 4:7-12.
[7] Terlizzi, M. E.; Gribaudo, G. and Maffei, M. E. (2017). UroPathogenic Escherichia coli (UPEC) Infections: Virulence Factors, Bladder Responses, Antibiotic, and Non-Antibiotic Antimicrobial Strategies. Front Microbiol. 8(1566): 1-23.
[8] Zowawi, H. M.; Harris, P. N. A.; Roberts, M. J.; Tambyah, P. A.; Schembri, M. A; Pezzani, M. D.; Williamson, D. A. and Paterson, D. L. (2015).The Emerging Threat of Multidrug-Resistant Gram-Negative Bacteria in Urology. Nat Rev Urol. 1-15.
[9] Leboffeem, M. J. and pierce, B.E. (2011). AphotographicAtls of the Microbiology Laboratory ,4thed , USA. pp:58-64,74-78,96-99,153.
[10] Kaye, Keith S. ; Engemann, John J.; Fraimow , Henry S. and Abrutyn, E. (2004). Pathogens resistant
to antimicrobial agents : epidemiology, molecular mechanisms and clinical management; Infect. Dis. Clin. N. Am.; 18:467-511.
[11] Dionisio, F.; Matic, I.; Radman, M.; Rodrigues, O. R. and Taddei, F. (2002). Plasmids spread very fast in heterogenous bacterial communities. Genetics. 162:1525-1532.
[12] Fishbach, M.A.; Walsh, C.T. (2009). Antibiotics for emerging pathogens . Science 325: 1089-1093.
[13] Oancea, Simona and Stoia, Mihaela.(2010). Antibiotic resistance of bacterial pathogens: The
magnitude of the problem from two perspectives – Romanian and worldwide ,15, 5:5519-5529 .
[14] Yoneyama , Hiroshi and Katsumata , Ryoichi. (2006). Antibiotic resistance in Bacteria and it Future for novel Development , biosci. Biotechno. Biochem, 70(5),1060-1075 .
[15] Souza, E.L.; Stamford, L.M.; Lima, E.D.; Trajano, V.n. and Filho, J.M. (2005). Antimicrobial
effectiveness of spices : an approach for use in food conservation system . Brazilian Archives of Biology and Technology -An International Journal , 48(4): 549-558.
[16] Cheesbrough, M. (2012). District Laboratory Practice in Tropical Countries. Second edition update (part2), Cambridge university press: India.
[17] Leboffeem, M. J. and pierce, B.E. (2011). AphotographicAtls of the Microbiology Laboratory ,4thed ,USA. pp:58-64,74-78,96-99,153.
[18] Jorgensen, J. H., Pfaller, M. A., Carroll, K.C. (2015). American Society for Microbiology. Manual of clinical microbiology. 11th ed. Washington, DC: ASM.423.
[19] Al-Kareemi, K. K., (2012). Inhibitory Effect of Parsley (Petroselinum Crispum) Juice against Some Urinary Pathogens in Vitro. The Iraqi Postgraduate Journal Medical. 11(3):336-342.
[20] Al-thoyni, Amina.N , Sanar.S and Sabah.M .(2008). Effect of water and alcoholic extracts for carnation and black pill and appointment of bezel in growth of some pathogens. Al-Nahrain University Journal.Vol 11, No.2.
[21] Sabah, J.,(2011). Antimicrobial activity of NutmegExtracts Against Staphylococcusaureus and Escherichia coli. J. Institute of Medical Technology. Baghdad. 8(2): 189-199.
[22] Rana, M. A. A., Sawsan, M. A. A., Khabat, A. A., and Wasam, A., (2014). Antibacterial activity of parsley and celery aqueous extract on isolated bacteria from children UTI in Erbil city. Interanational journal of advanced.2(9):895-903.
[23] Boas C. L . van der putten, Daniel Remondini, Giovanni Pasquini, Victoria A.Janes, Sebastien
Matamoros and Constance schultsz. (2019). Quantifing the Contribution of four resistance mechanisms to Ciprofloxacin MIC in Escherichia coli :a systematic review. University of Amsterdam, Department of Microbiology, chemother;74:298- 310.
[24] Wafaa, S. A.W., and Bashair, S.H., (2015). Anti- Virulence Effect of Pomegranate Peel Extracts on Most common urinary tract infection pathogens in pregnant women,Journal of Contem poray Medical Science. College of science and technology.(1)4:7-12.
[25] Chen, W. P.; and Kuo, T.T. (1993). A simple and rapid method for the preparation of gram-negative bacteria genomic DNA. Nucleic acids research, 21 (9), 2260. ISO 690.
[26] Sambrook, J., and Russel, D.W. (2001). Molecular cloning, Alaboratory manual (3ed). Cold Spring Harbor laboratory press, New York.
[27] Weigand , F., Baum, M. and Udupa , S. (1993). DNA molecular marker techniques, technical manual. No.20. International Center for Agricultural Research in the Dry Area(ICARDA). Aleppo, Syria.
[28] Radi A. M, Aziza A. A, Sherif Z and Ahmed H. (2017). Evaluation of genetic polymorphism, genomic templet stability, condition facter and hemato-biochemical level during Nile tilapia.
[29] Taspinar, MS, Agar, G., Yildirim, N., Sunar, S., Aksakal, O., Bozari, S. (2009). Journal of food Agriculture and Environment, 7 (3 & 4): 857-680.
[30] Widad ,S,J, Al-Zirjawi & Saad ,S, S.(2016). Serotyping and molecular detection of some resistance genes of Escherichia coli isolated from urinary tract infection children in Nasiriyah City. University of Thi-Qar Journal,Volume 11, Issue 3, page 1-20.
[31] Seifu, WD; Gebissa, AD. (2018). Prevalence and antibiotic susceptibility of Uropathogens from cases of urinary tract infections (UTI) in Shashemene referral hospital, Ethiopia.BMC Infections Diseases.; 18:30.
[32] Almazroo'e, Othman ,S,M.(2017).Cumulative effect of aqueous and alcoholic extracts of radish and
ginger plants on some bacteria isolated from urinary tract infections in Tikreet city. Submitted Master in Biology/ Microbiology/ Tikrit University.
[33] P. Ponnusamy & Ramanathan Nagappan. (2013).Extended Spectrum Beta -Lactamase, Biofilm-
producing Uropathogenic Pathogens and Their Antibiotic Susceptibility Patterns from Urinary Tract
Infection- An Overview. Published 2013,Biology.
[34] Hegazy,E .E. Alam El-Din, R. A.; Amin, A.M.; Mahgoub, F.M. and El-Gamal, S.A. (2018). Microbiological Profile of Urinary Tract Infections with special Refrence to Antibiotic Susceptibility Pattern of Escherichia coli Isolates. Int J Curr Microbiol App Sci. 7(2): 911-920.
[35] Raeispour, M. and Ranjbar. R. (2018). Antibiotics Resistance, Virulence Factors and Genotyping of Uropathogenic Escherichia coli Strains. Antimicrob Resist Infect Control. 7 (118): 1-9.
[36] Carroll, K. C., Butel, J. S., Morse, S. A. (2016). Jawetz Melnick & Adelbergs Medical Microbiolog . 27 ed. McGraw Hill Professional.
[37] Brown, A. E., Smith, H. A. (2017). Benson´ s. Microbiological applications: laboratory manual in general microbiology .14th ed. McGraw-Hill. New York.
[38] Tille, P.M. (2017). Baily And Scott’s Diagnostic Microbiology. 41thed. Elsevier, Inc. China. 1115pp.
[39] CLSI (2018). Performance Standards for Antimicrobial Susceptibility Testing; Twenty-Fifth Informational Supplement. Approvedstandard M100-S25. CLSI, Wayne, PA.
[40] Hilan , Saba, F, .(2019).Inhibitory Effect of Biological Products Extracted from some Cyanobacteria species on Multi Antibiotic Resistant Bacteria. Submitted Master, Collage of Science/ Department of Biology/ Tikrit University.
[41] Janssen, H., Janssen, I., Cooper, P., Kainyah, C., Pellio, T., Quintel, M., Schulze, M. H. (2018). Antimicrobial-Resistant Bacteria in Infected Wounds, Ghana, 2014. Emerging infectious diseases, 24(5), 916-919.
[42] Patel, H. B., Soni, S. T., Bhagyalaxmi, A., Patel, N. M. (2019). Causative agents of urinary tract infections and their antimicrobial susceptibility patterns at a referral center in Western India: An audit to help clinicians prevent antibiotic misuse. Journal of Family Medicine and Primary Care, 8(1),154-159.
[43] Rajabnia, M., Forghani, M. S., Hasani, S., Bahadoram, M., Mohammadi, M., &Barahman, M. (2019). Prevalence and antibiotic resistance pattern of extended spectrum beta lactamase producing Escherichia coli isolated from urinary tract infection. Journal of Renal Injury Prevention, 8(2).
[44] Chowdhury, S., Parial, R. (2015). Antibiotic susceptibility patterns of bacteria among urinary tract infection patients in Chittagong, Bangladesh. SMU Medical Journal, 2(1), 114-127.
[45] Al-saadi, Zahraa, H,A.(2019).Phenotypic and molecular detection of Escherichia coli efflux pumps from UTI patients. Submitted Master, Collage of Science/ Ibn Al-Haitham / Department of Biology/ University of Baghdad.
[46] Tajbakhsh, E.; Ahmadi, P. Abedpour-Dehkordi, E.; Arbab-Soleimani, N. and Khamesipour, F.(2016).
Biofilm Formation, Antimicrobial Susceptibility, Serogroups and Virulence Genes of Uropathogenic E. coli Isolated from Clinical Samples in Iran. Antimicrob Resist Infect Control. 5(11): 1-8.
[47] Al-Tikrity, I. A. R. (2016). Detection of some Virulence Genes in Escherichia coli Isolated fromPatients with Urinary Tract Infection in Erbil City. Ph.D. thesis,College of Science, Tikrit University.
[48] Kurutepe, S. S., and Surucuoglu, (2014). Increasing antimicrobial resistance in Escherichia coli isolates from community-acquired urinary tract infections during 1998-2003 in Turkey. Jpn. J.Infect. Dis. 58:159-161.
[49] Linuma, Y., (2007). Infection control strategies for antimicrobial resistance. Nippon Rinsho. 65:175-184.
[50] Chen CJ1, Huang YC. (2014).New epidemiology of Staphylococcus aureus infection in Asia. Clin
Microbiol Infect. Jul;20(7):605-23. [51] Jarlier V, Carlet J, McGowan J, et al.(2012).
Participants of the 3rd World Healthcare-Associated
Infections Forum Priority actions to fight antibiotic resistance: results of an international meeting. Antimicrob Resist Infect Control. 1(1):17. [52] Furtula V1, Farrell EG, Diarrassouba F, Rempel H, Pritchard J, Diarra MS.(2010). Veterinary pharmaceuticals and antibiotic resistance of Escherichia coli isolates in poultry litter from
commercial farms and controlled feeding trials. Poult Sci. Jan;89(1):180-8. [53] Ferri M, Ranucci E, Romagnoli P, Giaccone V. (2017). Antimicrobial resistance: A global emerging threat to public health systems. Crit Rev Food Sci Nutr. 2017 Sep 2;57(13):2857-2876. [54] Tortora, G.J; Funke, B.R. and Case, C.L. (2004) .Microbiology an introduction. Eighth edition; pearson Benjamin Cummings, Sanfrancisco, Boston, New york San Francisco.
[55] Rudresh SM, Nagarathnamma T. (2011). Extended spectrum β-lactamase producing Enterobacteriaceae & antibiotic co-resistance. Indian J Med Res . Jan;133:116-118.
[56] Jisun K., Bora Sh., Chulwoo P., and Woojun Park (2017).Indole-Induced Activities of β-Lactamase and Efflux Pump Confer Ampicillin Resistance in Pseudomonas putida KT2440 2017; 8: 433. [57] Jassim SA, Limoges RG.(2014). Natural solution to antibiotic resistance: bacteriophages ‘The Living Drugs’ World J Microbiol Biotechnol. 30(8):2153–2170. [58] Diarra MS, Malouin F.(2014). Antibiotics in Canadian poultry productions and anticipated alternatives.Front Microbiol. Jun 17;5:282.
[59] Wafaa, S. A.W., and Bashair, S.H., (2015). Anti- Virulence Effect of Pomegranate Peel Extracts on
Most common urinary tract infection pathogens in pregnant women,Journal of Contem poray Medical Science. College of science and technology.(1)4:7-12.
[60] Al-Doori, A.A , Awatef Saber Jasem and Adnan F. Al-Azzawie .(2020). Molecular Assessment Effect of Nd: YagLaser on Pseudomonas aeurginosa Bacteria Using RABD Marker. Tikrit University College of Science. Department of Physics.
[61] Taylor, V. E.; Brady, L.R. and Robers, J.E., 1988, Pharmacognosy. 3ed.Lea and Febiger. Philadelphia.
[62] Ismail T., Sistili P., and Akhtar S. (2012). Pomegranante Peel and Fruit Extracts: A Review of Potential Anti-Inflammatory and Anti-Infective Effects Journal of Ethnopharmacology /143/ Pages 397-405. [63] Chauhan, G., M., Sharma, H., Kharkwal, and A. Varma. 2010. Pharmacognostic, preliminary phytochemical studies and anticancerous potential of Trigonella Foenum- Graecum. Inter. J. Pharm. Sci.: 350-60.
[64] Mishra .A.K. and Vinit, K.M.(2012). Field survy for some fungal disease on egg plant. International
Multidisciplinary Research Journal. Vol.(2). No.(9).
Pp:23.
[65] Bkhari , F.M (2009). Antifungul activity of some medicinal plants used in Jeddah, Saudia Arabia. Mycophat . Vol.(7).No. (1). Pp. 51-57.
[66] Atienzar, F. A. Conradi, M. Evenden, A. J., Jha, A. N. and Depledge, M.H.(1999). Qualitative assessment of genotoxicity using random amplified
polymorphic DNA: comparison of genomic template stability with key fitness arameters in daphnia magna exposed to benzo [a]pyrene. Environ. Toxicol. Chem.18:2275-2282.
[67] Ginchner, T., Zindar, I., Szakova, J. (2008). Evaluation of DNA damage and mutagenecity induced by lead in tobacco plants. Mut Res; 652: 186-190.
[68] Ahmad, M., Ahmad, M., Alshamali, b. (2010). No significant Cytogenetic Effects in Cultured Human Lymphocytes Exposed to Cell Phones Radiofrequencies (900MHz and 1800MHz). Jordan Journal of Biological Sciences. Volume 3, Number 1, January. Pages 21 – 28.
[69] Amjady, F., Golestani, E. B., Karimi, F., Sevda, T and Seyed, S. P. (2016). An Investigation of the Effect of Copper Oxide and Silver Nanoparticles on E. Coli Genome by Rapd Molecular Markers. J. Adv Biotechnology & Microbiology , Vol .1 , No. 2 , pp. 1-6.
[70] Gherbawy , Y.A. (2001). Use of RAPD –PCR to characterize Eurotium strains isolated from date fruits J. Cytologia , Vol.66 , pp. 349-356.
[71] Hadrys, H., Balick, M., Schierwater, B. (1992). Applications 22.of random amplified polymorphic DNA (RAPD) in molecular ecology. Mol Ecol; 1: 55-63.