ARTICLE INFO

Article Type

Original Research

Authors

Habibi   M. (1)
Asadi Karam   M.R (*)
Mohammadzadeh   A. (2)






(*) Department of Molecular biology, Pasteur Institute of Iran, Tehran, Iran
(1) Department of Molecular biology, Pasteur Institute of Iran, Tehran, Iran
(2) Department of Microbiology, School of Medicine, Gonabad University of Medical Sciences, Gonabad, Iran

Correspondence

Address: Department of Molecular biology, Pasteur Institute of Iran, Tehran, Iran
Phone: +98 (21) 66953311
Fax: +98 (21) 66370573
m_asadi12@yahoo.com

Article History

Received:  July  26, 2018
Accepted:  September 22, 2018
ePublished:  October 10, 2018

BRIEF TEXT


Urinary tract infection refers to a set of infections that various parts of the urinary tract are involved and these areas are infected [1].

... [2-4]. Various studies have shown that bacterial strains of Escherichia coli have always been the most common and important cause of these infections among the causative agents of urinary tract infections [5]. ... [6, 7]. Antimicrobial resistance between these strains is increasing, and this could lead to huge dilemmas in the future for the treatment of these infections [10-8]. ... [11, 12]. Integrons, as a movable genetic element, play an important role in the transmission of antibiotic resistance among gram-negative bacteria, including E. coli. These factors alone can not be transmitted, but fused in plasmids or transposons, they can easily be transmitted through horizontal transmission between bacterial strains [13,14]. .... [15-18].

The aim of the present study was detection of integrons and their relationships with antibiotic resistance in E. coli. isolates.

This research is an experimental study.

This experimental study was performed on E. coli. isolates isolated from urinary tract infection in patients referred to several hospitals in Tehran in 2017.

A total of 150 E. coli isolates of the urinary tract were collected from women with urinary tract infection from Tehran hospitals aged between 18 and 60 years old.

In the present study, only urinary cultures with urinary tract infections infected by E. coli were selected and a total of 150 E. coli isolates isolates were obtained. To ensure proper identification of the collected isolates, the diagnostic tests for E. coli were repeated in the molecular biology laboratory of the Pasteur Institute of Iran. Then, the resistance of isolates to different classes of antibiotics, including beta-lactams, carbapenems, fluoroquinolones, aminoglycosides, and trimethoprim-sulfamethoxazole (25 µg) was measured by disc diffusion method and according to the Clinical & Laboratory Standards Institute (CLSI) guidelines [19]. In this study, the E. coli strain ATCC25922 was used as a control strain. In the next step, identification of ESBL-carrying isolates was carried out using phenotypic combined disk diffusion test (CDDT) based on the CLSI protocol. In this method, the cefotaxime disc plus cefotaxime+clavulanic acid combined disc as well as ceftazidime disc plus ceftazidime+clavulanic acid combined disc were used [19]. The standard Klebsiella pneumoniae strain ATCC70603 was used as a positive control. In the next step, the replication of the integrin I (intI) genes was accomplished by polymerase chain reaction (PCR) [12]. The relationship between integron resistance genes with antibiotic resistance and ESBL production was investigated using chi-square test. In addition, SPSS 19 software was used.

The isolates showed the highest antibiotic resistance to amoxicillin (71.3%), and the least resistance was related to the antibiotics imipenem (0%), amikacin (2%) and morphenem (3.3%), respectively. the following values obtained for resistance to other antibiotics: trimethoprim-sulfamethoxazole (64%), cefotaxime (55.3%), ceftazidime (52.7%), ciprofloxacin (52%), norfloxacin (46.7%) and gentamycin (19.3%). According to antimicrobial resistance pattern, 70 isolates showed antibiotic resistance to at least three antibiotic families and were considered as multi-antibiotic resistance (MDR) strains (isolates resistant to 3 or more antibiotic classes) (Table 1). According to Table 2, resistance to all antibiotics in ESBL-carrying strains was higher than non-ESBL-carrying strains. Also, there was a significant relationship between the ESBL-carrying strains and the resistance to antibiotics, including beta-lactam, trimethoprim-sulfamethoxazole, ciprofloxacin, norfloxacin and gentamicin (p<0.05). All ESBL-carrying strains showed resistance to amoxicillin and cefotaxime. On the other hand, there was a significant relationship between ESBL-carrying strains and MDR resistance (p<0.001). The integron size is about 491 bp (Figure 1). The highest resistance in the integrin-carrying isolate was observed in amoxicillin and trimethoprim-sulfamethoxazole antibiotics (Table 3). Resistance to amikacin was observed only among integrin-carrying isolate, while no integrin-carrying isolate was found resistant to meropenem. Also, there was a significant relationship between the presence of the integron and resistance to amoxicillin, trimethoprim-sulfamethoxazole, gentamycin and amikacin (p<0.05). The distribution of isolates with integrase gene with the antibiotic resistance pattern is given in Table 4. Of the 34 isolates containing integrative gene, 31 isolates showed resistance to at least one antibiotic, and only 3 isolates showed reaction to all antibiotics tested (Table 4). There was a significant correlation between integron Class I and MDR resistance (p=0.02), and it was also observed that among the 56 isolates carrying ESBL, 18 isolates had integron Class I (p=0.043).

According to the present study, the highest resistance observed in amoxicillin. However, in other studies, the high resistance of E. coli strains was observed in beta-lactam family antibiotics, including ampicillin and amoxicillin [10, 20, 21]. Consistent with this study, Imipenem was reported as one of the most effective antibiotics against urinary tract infections [10]. ... [22, 23]. Various studies in Iran have shown that the percentage of E. coli isolates isolated from urinary tract infection are resistant to aminoglycosylated antibiotics in Jahrom (11.7%) [24] to 76.6% in Zahedan [25], which is higher than the observed aminoglycoside resistance observed in this study. ... [26, 27]. In a review by Hadi Far et al. in Iran, the average prevalence of E. coli MDR strains in different parts of Iran was reported about 50%, which was lower than the median MDR in isolates studied in the present study [20]. .... [28, 29]. In this study, 37.3% of isolates were able to carry ESBL, and according to studies, the prevalence of ESBL strains is from 2.4 in Tehran to 62.7% in Zahedan [10]. ... [30-32]. In the present study, only 22.7% of tested isolates were Class I integron-carrying isolates. In other studies in Iran, it is ranged from 6.2% in Shiraz [26] to 52% in Yasuj, and 97% in the north. ... [34-38].

More studies are suggested to make more favorable conditions for preventing antibiotic resistance.

None declared.

The significant resistance was observed for most antibiotics tested with ESBL production, which can be due to the class I integrons.

We are grateful to the Pasteur Institute for supporting this project.

None declared.

None declared.

This study was funded by the Pasteur Institute of Iran.

TABLES and CHARTS

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CITIATION LINKS

[1]Wright KJ, Seed PC, Hultgren SJ. Uropathogenic Escherichia coli flagella aid in efficient urinary tract colonization. Infect Immun. 2005;73(11):7657-68.
[2]Gupta K, Hooton TM, Naber KG, Wullt B, Colgan R, Miller LG, et al. International clinical practice guidelines for the treatment of acute uncomplicated cystitis and pyelonephritis in women: A 2010 update by the Infectious Diseases Society of America and the European Society for Microbiology and Infectious Diseases. Clin Infect Dis. 2011;52(5):e103-20.
[3]Nicolle LE. Uncomplicated urinary tract infection in adults including uncomplicated pyelonephritis. Urol Clin North Am. 2008;35(1):1-12.
[4]Salvatore S, Salvatore S, Cattoni E, Siesto G, Serati M, Sorice P, et al. Urinary tract infections in women. Eur J Obstet Gynecol Reprod Biol. 2011;156(2):131-6.
[5]Dhakal BK, Kulesus RR, Mulvey MA. Mechanisms and consequences of bladder cell invasion by uropathogenic Escherichia coli. Eur J Clin Invest. 2008;38 Suppl 2:2-11.
[6]Vachhani AV, Barvaliya M, Naik V, Jha P, Tripathi C. Effectiveness and tolerability of short course co-trimoxazole, norfloxacin and levofloxacin in bacteriological cure of uncomplicated urinary tract infection in outpatient setting, an open label, parallel group, randomized controlled trial. Infez Med. 2015;23(2):155-60.
[7]Fasugba O, Gardner A, Mitchell BG, Mnatzaganian G. Ciprofloxacin resistance in community- and hospital-acquired Escherichia coli urinary tract infections: A systematic review and meta-analysis of observational studies. BMC Infect Dis. 2015;15:545.
[8]Walker E, Lyman A, Gupta K, Mahoney MV, Snyder GM, Hirsch EB. Clinical management of an increasing threat: Outpatient urinary tract infections due to multidrug-resistant uropathogens. Clin Infect Dis. 2016;63(7):960-5.
[9]Sanchez GV, Babiker A, Master RN, Luu T, Mathur A, Bordon J. Antibiotic resistance among urinary isolates from female outpatients in the United States in 2003 and 2012. Antimicrob Agents Chemother. 2016;60(5):2680-3.
[10]Alizade H. Escherichia coli in Iran: An overview of antibiotic resistance, a review article. Iran J Public Health. 2018;47(1):1-12.
[11]Tong YQ, Sun M, Hu C, Zhao D. Plasmid transfer capacities of multi-resistant UPEC clinical isolates in biofilms. Biomed Res. 2017;28(5):2364-9.
[12]Paniagua-Contreras GL, Monroy-Pérez E, Rodríguez-Moctezuma JR, Domínguez-Trejo P, Vaca-Paniagua F, Vaca S. Virulence factors, antibiotic resistance phenotypes and O-serogroups of Escherichia coli strains isolated from community-acquired urinary tract infection patients in Mexico. J Microbiol Immunol Infect. 2017;50(4):478-85.
[13]Kaushik M, Kumar S, Kapoor RK, Virdi JS, Gulati P. Integrons in Enterobacteriaceae: Diversity, distribution and epidemiology. Int J Antimicrob Agents. 2018;51(2):167-76.
[14]Fluit AC, Schmitz FJ. Resistance integrons and super-integrons. Clin Microbiol Infect. 2004;10(4):272-88.
[15]Mazel D. Integrons: Agents of bacterial evolution. Nat Rev Microbiol. 2006;4(8):608-20.
[16]Rowe-Magnus DA, Mazel D. Integrons: Natural tools for bacterial genome evolution. Curr Opin Microbiol. 2001;4(5):565-9.
[17]Lina TT, Khajanchi BK, Azmi IJ, Islam MA, Mahmood B, Akter M, et al. Phenotypic and molecular characterization of extended-spectrum beta-lactamase-producing Escherichia coli in Bangladesh. PLoS One. 2014;9(10):e108735.
[18]Barguigua A, El Otmani F, Talmi M, Bourjilat F, Haouzane F, Zerouali K, et al. Characterization of extended-spectrum β-lactamase-producing Escherichia coli and Klebsiella pneumoniae isolates from the community in Morocco. J Med Microbiol. 2011;60(Pt 9):1344-52.
[19]Patel JB, Cockerill III FR, Bradford PA, Eliopoulos GM, Hindler JA, Jenkins SG, et al. M100-S25: Performance standards for antimicrobial susceptibility testing, twenty-fifth informational supplement [Internet]. Wayne PA: Clinical and Laboratory Standards Institute; 2015 [cited 2017 Sep 20]. Available from: http://file.qums.ac.ir/repository/mmrc/CLSI2015.pdf
[20]Hadifar S, Moghoofei M, Nematollahi S, Ramazanzadeh R, Sedighi M, Salehi Abargouei A, et al. Epidemiology of multidrug resistant uropathogenic Escherichia coli in Iran: A systematic review and meta-analysis. Jpn J Infect Dis. 2017;70(1):19-25.
[21]Terlizzi ME, Gribaudo G, Maffei ME. UroPathogenic Escherichia coli (UPEC) infections: Virulence factors, bladder responses, antibiotic, and non-antibiotic antimicrobial strategies. Front Microbiol. 2017;8:1566.
[22]Enne VI, Livermore DM, Stephens P, Hall LM. Persistence of sulphonamide resistance in Escherichia coli in the UK despite national prescribing restriction. Lancet. 2001;357(9265):1325-8.
[23]Mehdipour Moghaddam MJ, Mirbagheri AA, Salehi Z, Habibzade SM. Prevalence of class 1 integrons and extended spectrum beta lactamases among multi-drug resistant Escherichia coli isolates from North of Iran. Iran Biomed J. 2015;19(4):233-9.
[24]Asadi S, Kargar M, Solhjoo K, Najafi A, Ghorbani Dalini S. The association of virulence determinants of uropathogenic Escherichia coli with antibiotic resistance. Jundishapur J Microbiol. 2014;7(5):e9936.
[25]Shayan S, Bokaeian M, Shahraki S. Prevalence and molecular characterization of AmpC-producing clinical isolates of Escherichia coli from Southeastern Iran. Microb Drug Resist. 2014;20(2):104-7.
[26]Farshad S, Japoni A, Hosseini M. Low distribution of integrons among multidrug resistant E. coli strains isolated from children with community-acquired urinary tract infections in Shiraz, Iran. Pol J Microbiol. 2008;57(3):193-8.
[27]Gholipour A, Soleimani N, Shokri D, Mobasherizadeh S, Kardi M, Baradaran A. Phenotypic and molecular characterization of extended-spectrum β-lactamase produced by Escherichia coli, and Klebsiella pneumoniae isolates in an educational hospital. Jundishapur J Microbiol. 2014;7(10):e11758.
[28]Dash SK, Chakraborty SP, Mandal D, Roy S. Isolation and characterization of multi drug resistant uropathogenic Escherichia coli from urine sample of Urinary tract infected patients. Int J Life Sci Pharm Res. 2012;2(1):L25-39.
[29]Sanchez GV, Baird AM, Karlowsky JA, Master RN, Bordon JM. Nitrofurantoin retains antimicrobial activity against multidrug-resistant urinary Escherichia coli from US outpatients. J Antimicrob Chemother. 2014;69(12):3259-62.
[30]Bali EB, Açık L, Sultan N. Phenotypic and molecular characterization of SHV, TEM, CTX-M and extended-spectrum β-lactamase produced by Escherichia coli, Acinobacter baumannii and Klebsiella isolates in a Turkish hospital. Afr J Microbiol Res. 2010;4(8):650-4.
[31]Sorlózano A, Gutiérrez J, Fernández F, Soto MJ, Piédrola G. A preliminary study on the presence of Extended-Spectrum Beta-Lactamases (ESBL) in clinical isolates of Escherichia coli in Granada (Spain). Ann Microbiol. 2004;54(2):99-104.
[32]Franiczek R, Dolna I, Krzyżanowska B, Szufnarowski K, Kowalska-Krochmal B. Conjugative transfer of multiresistance plasmids from ESBL-positive Escherichia coli and Klebsiella spp. clinical isolates to Escherichia coli strain K12 C600. Adv Clin Exp Med. 2007;16(2):239-47.
[33]Khoramrooz SS, Sharifi A, Yazdanpanah M, Malek Hosseini SAA, Emaneini M, Gharibpour F, et al. High frequency of class 1 integrons in Escherichia coli isolated from patients with urinary tract infections in Yasuj, Iran. Iran Red Crescent Med J. 2016;18(1):e26399.
[34]Salem MM, Muharram M, Alhosiny IM. Distribution of classes 1 and 2 integrons among multi drug resistant E. coli isolated from hospitalized patients with urinary tract infection in Cairo, Egypt. Aust J Basic Appl Sci. 2010;4(3):398-407.
[35]Al-Assil B, Mahfoud M, Hamzeh AR. First report on class 1 integrons and Trimethoprim-resistance genes from dfrA group in Uropathogenic E. coli (UPEC) from the Aleppo area in Syria. Mob Genet Elements. 2013;3(3):e25204.
[36]Machado E, Cantón R, Baquero F, Galán JC, Rollán A, Peixe L, et al. Integron content of extended-spectrum-beta-lactamase-producing Escherichia coli strains over 12 years in a single hospital in Madrid, Spain. Antimicrob Agents Chemother. 2005;49(5):1823-9.
[37]Bonnet R. Growing group of extended-spectrum beta-lactamases: The CTX-M enzymes. Antimicrob Agents Chemother. 2004;48(1):1-14.
[38]Jacoby GA, Sutton L. Properties of plasmids responsible for production of extended-spectrum beta-lactamases. Antimicrob Agents Chemother. 1991;35(1):164-9.