بررسی شیوع ژن‌های qnr در انتروباکتریاسه‌های جداسازی شده در فاضلاب شهری و بیمارستانی

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانش‌آموخته کارشناسی ارشد، گروه میکروبیولوژی، دانشکده علوم، دانشگاه آزاد اسلامی واحد کرج، کرج، ایران

2 استادیار، گروه میکروبیولوژی، دانشکده علوم، دانشگاه آزاد اسلامی واحد کرج، کرج، ایران

3 استادیار، گروه حفاظت محیط‌زیست، پژوهشگاه صنعت نفت، تهران، ایران

چکیده

امروزه مقاومت به آنتی‌بیوتیک‌ها در باکتری‌ها به‌دلیل استفاده نادرست رو به افزایش است که این امر برای درمان بیماری‌ها یک تهدید به‌حساب می‌آید. فاضلاب‌های بیمارستانی، پتانسیل زیادی برای ورود باکتری‌ها و ژن‌های مقاوم به محیط‌زیست دارند. همچنین سیستم‌های تصفیه کنونی نیز کارایی زیادی در حذف و یا کاهش آنتی‌بیوتیک‌ها ندارند که این امر باعث افزایش باکتری‌ها با ژن‌های مقاومت به آنتی‌بیوتیک‌ها در محیط می‌شود. هدف از این پژوهش بررسی شیوع ژن‌های پلاسمیدی qnr در جدایه‌های انتروباکتریاسه مقاوم به کوئینولون از فاضلاب‌های شهری و بیمارستانی شهر کرج بود. تعداد 99 جدایه انتروباکتریاسه از پساب بیمارستانی و شهری استان البرز در بهار 1398 جداسازی شدند. آزمون‌های بیوشیمیایی به‌منظور شناسایی آنها انجام شد و جدایه‌ها از نظر الگوی مقاومت به آنتی‌بیوتیک‌های سیپروفلوکساسین، لووفلوکساسین، افلوکساسین، نورفلوکساسین و نالیدیکسیک اسید به روش کربی‌ بایر بررسی و جدایه‌هایی که بیشترین مقاومت به کوئینولون‌ها را داشتند. برای حضور ژن‌های qnr A، qnr B و qnr S به روش PCR بررسی شدند. در میان ارگانیسم‌های کشت شده اشریشیاکلی شایع‌ترین ارگانیسم بود و بعد از آن اشریشیاکلی (غیر فعال) و سیتروباکتر فروندی قرار داشتند. بیشترین مقاومت در بین جدایه‌ها به آنتی‌بیوتیک نالیدیکسیک اسید مشاهده شد. بیشتر جدایه‌ها حاوی ژن qnr A (61 درصد) بودند. ژن‌های qnr B و qnr S به‌ترتیب در 31 درصد و 8 درصد از جدایه‌ها یافت شد. یک جدایه حاوی هر دو ژن qnr B و qnr A بود. بیشترین جدایه‌های حاوی ژن qnr A از پساب بیمارستانی و بیشترین جدایه‌های حاوی ژن qnr B از پساب شهری جداسازی شدند. مقاومت آنتی‌بیوتیکی امروزه باید به‌عنوان یک آلودگی زیست‌محیطی در نظر گرفته شود و کیفیت تصفیه فاضلاب در کاهش این عوامل باید ارزیابی شود. همچنین مصرف خودسرانه آنتی‌بیوتیک‌ نیز باید متوقف شوند.

کلیدواژه‌ها


عنوان مقاله [English]

Frequency Assay of qnr Genes in Enterobacteriaceae Isolated from Hospital and Urban Wastewaters in Karaj City

نویسندگان [English]

  • Shirin Rezakhani 1
  • Azam Haddadi 2
  • Mahmoud Shavandi 3
1 MSc. Graduate Student, Dept. of Microbiology, Karaj Branch, Islamic Azad University, Karaj, Iran
2 Assist. Prof., Dept. of Microbiology, Karaj Branch, Islamic Azad University, Karaj, Iran
3 Assist. Prof., Ecology and Environmental Pollution Control Research Group, Research Institute of Petroleum Industry, Tehran, Iran
چکیده [English]

In recent years, because of excessive and unregulated use of antibiotics, the threat of acquisition of antibiotic resistance by pathogens is growing. Hospitals are hotspots for antimicrobial-resistant bacteria (ARB) and will be ejected from hospitals via wastewater systems. The aim of this study was the frequency assay of qnr plasmid’s genes in quinolone resistant Entrobacteriaceae strains isolated from urban and hospital wastewaters. A total of 99 Entrobacteriaceae strains were isolated from urban and hospital wastewater in Alborz province during the spring of 2019. Bacterial strains were identified by standard microbiological and biochemical tests. The antimicrobial susceptibility test to ciprofloxain, levofloxacin, ofloxacin, norfloxacine and nalidxic acid was determined by Kirby-Bauer method and the frequency of quinolone resistance genes (qnrA, qnrB, qnrS) was investigated by PCR. Among the organisms cultured, Escherichia coli was the most common organism followed by Escherichia coli (inactive) and Citrobacter freundii. The most antibiotic resistance was observed against nalidixic acid.  Most of the isolates (61%) harboured the qnrA gene. qnrB and qnrS genes were found in 31% and 8% of isolates respectively. One isolate has both qnrA and qnrB genes. Most of the strains that contain qnrA and qnrB genes were isolated from hospital and urban wastewater respectively. Antimicrobial resistance should now be seen as an environmental pollutant and new wastewater treatment processes must be assessed for their capability in eliminating antimicrobial-resistant bacteria, especially from hospital effluents. Also, unregulated use of antibiotics should be stopped.

کلیدواژه‌ها [English]

  • Quinolones
  • Qnr Genes
  • Hospital Wastewater
  • Antibiotic Resistance
Aali, R., Hosseinpoor, S., Shahryari, A. & Mirhosseini, S. H. 2017. Diversity of genes coding of antibiotic resistance in municipal westewaters. Rahavard Salamat Journal, 2(3), 1-14. (In Persian)
Aali, R., Nikaeen, M., Hatamzadeh, M., Moazeni, M., Khanahmad, H. & Shahryari, A. 2016. The role of hospital wastewaters in dissemination of antibiotic resistant bacteria & resistance genes to the environment. Journal of Environmental Health Enginering, 3(3), 239-248.
Aali, R., Nikaeen, M., Khanahmad, H., Hejazi, Z., Kazemi, M. & Hassanzadeh, A. 2014. Occurrence of tetracycline resistant bacteria and resistance gene (fetW) in hospital and municipal wastewaters. Fresenius Environmental Bulletin, 23(10A), 2560-2566. (In Persian)
Abbassi Ghozzi, I., Jaouani, A., Hammami, S., Martinez Urtaza, J., Boudabous, A. & Gtari, M. 2012. Molecular analysis and antimicrobial resistance of Salmonella isolates recovered from raw meat marketed in the area of Grand Tunis,Tunisia. Pathologie Biologie, 60, 49-54.
Afshari, M., Haddadi, A. & Shavandi, M. 2020. Prevalence assay of antibiotic resistance in Enterobacteriaceae isolated from hospital and urban wastewaters in Karaj city. Journal of Water and Wastewater, 31(3), 68-76. (In Persian)
Badalians Gholikand, G. 2016. Comprehensive microbiology of water and wastewater. AIJ Publications, Tehran, 345-367. (In Persian)
Banjara, M. R., Sharma, A. P., Joshi, A. B., Tuladhar, N. R., Ghimire, P. & Bhatta, D. R. 2003. Surgical wound infections in patients of Tribhuvan university teaching hospital. Nepal Health Research Council, 3, 5-41.
Chegene Lorestani, R., Akya, A., Elahi, A. & Hamzavi, Y. 2017. Frequency of plasmid-mediated quinolone resistance genes in extended-spectum β-lactamase-producing escherichia coli. Journal of Mazandaran University of Medical Sciences, 27(151), 41-51. (In Persian)
Elmanama, A. A. and ElKichaoui, A. Y. & Mohsin M. 2006. Contribution of hospital wastewater to the spread of antibiotic resistance in comparison to non-health. Journal of Al-Aqsa University, 10(1), 108-121.
Faghri, J., Dehbanipour, R., Mobasherizadeh, S. & Maleki, N. 2016. Study of antibiotic resistance pattern and mutation in genes gyrA and parC of escherichia coli causing urinary tract infection. Avicenna Journal of Clinical Medicine (Scientific Journal of Hamadan University of Medical Sciences and Health Services), 23(2), 118-125. (In Persian)
Falah, F., Mirdehghan, S. A., Faramarzi, N., Eslami, G., Taheri, S., Navidinia, M., et al. 2009. Bacterial agents in patients with conjunctivitis in labbafinejad hospital in 2006. Research in Medicine, 23(2), 112-116. (In Persian)
Farshchian, M., Roshani, M. & Dehghanzadeh Reihani, R. 2015. Determination of antibiotic resistance pattern in bacteria isolated from municipal wastewater treatment plant. Journal of Mazandaran University of Medical Sciences, 25(126), 11-21. (In Persian)
Firoozeh, F., Zibaei, M. & Soleimani Asl, Y. 2014. Detection of plasmid-mediated qnr genes among the quinolone-resistant Escherichia coli isolates in Iran. The Journal of Infection in Developing Countries, 80(07), 818-822.
Gadage, D., Wankhade, A., Muley, V. V., Paralikar, A. V. & Bhore, A. V. 2014. Are inactive E.coli always commensals. Scholars Journal of Applied Medical Sciences, 2, 426-427.
Ghane, M. & Khanpour Zarenji, R. 2015. Detection of antibiotic resistant gram negative bacteria and plasmid profiling of multi-drug resistantes in hospital effluents. Medical Sciences, 24(4), 235-241. (In Persian)
Guardabassi, L., Wong, D. M. L. F. & Dalsgaard, A. 2002. The effects of tertiary wastewater treatment on the prevalence of antimicrobial resistant bacteria. Water Research, 36(8), 1955-1964.
Hadi, M., Shokoohi, R., Ebrahimzadeh Namvar, A., Karimi, M. & Solaimany Aminabad, M. 2011. Antibiotic resistance of isolated bacteria from urban and hospital wastewaters in Hamadan city. Iranian Journal of Health and Environment, 4(1), 105-114. (In Persian)
ISIRI 1011, Drinking water – Microbiological specifications, 1985. (In Persian)
ISIRI 4208, Water quality - Sampling for microbiological examination of water – Code of practice, 2016.
(In Persian)
Jeong, H. S., Bae, I. K., Shin, J. H., Jung, H. J., Kim, S. H., Lee, J. Y., et al. 2011. Prevalence of plasmid-mediated quinolone resistance and its association with extended-spectrum beta-lactamase and AmpC beta-lactamase in Enterobacteriaceae . The Korean Journal of Laboratory Medicine, 31(4), 257-264.
Jha, A. K., Singh, J. B. & Dutta, D. 2007. Microorganisms present in discharging otitis media in a group of patients in Kathmandu. Nepal Medical College Journal, 9(3), 196-198.
Katzung, B. G. 2017. Basic and clinical pharmacology. 14th Ed., McGraw-Hill Education.
Kim, M. H., Lee, H. J., Park, K. S. & Suh, J. T. 2010. Molecular characteristics of extended spectrum β-lactamases in Escherichia coli and Klebsiella pneumoniae and the prevalence of qnr in extended spectrum β-lactamase isolates in a tertiary care hospital in Korea. Yonsei Medical Journal, 51(5), 768-774.
Koneman, E. W. 2006. Koneman's color atlas and textbook of diagnostic microbiology. 6th Ed. Washington C:LWW.
Lavigne, J. P., Defez, C., Bouziges, N., Mahamat, A. & Sotto, A. 2007. Clinical and molecular epidemiology of multidrug-resistant Citrobacter spp. infections in a French university hospital. European Journal of Clinical Microbiology and Infectious Diseases, 26(6), 439-441.
Mansory Jamshidi, N., Pakzad, E., Tabaraee, B. & Hadadi, A. 2013. Frequency of qnr genes in escherichia coli strains resistant to quinolones isolated from Ilam Imam Khomani hospital and Tehran Milad hospital. Journal of Ilam University of Medical Sciences, 21(6), 16-22. (In Persian)
Misra, B., Gandham, N., Sardar, M., Ujagare, M., Angadi, K. & Vyawahare, C. H. 2012. High prevalence of multi-drug resistant Citrobacter spp. From tertiary car hospital, Pimpri, Pune. India. Journal of Pharmaceutical and Biomedical Sciences, 25(25), 158-163.
Moghbeli, M., Behnood, V. & Ranjbar, R. A. 2014. Study to determine antibiotic resistance and recognition qnr genes in shigella strains isolated from patients admitted to mofid's children medical center, Tehran. Journal of Microbial World, 7(1), 49-57. (In Persian)
Monzavi, M. T. 2010. Urban wastewater, wastewater treatment. Vol 2. University of Tehran Publishing Institute, Tehran, Iran. (In Persian)
Ng, C., Tay, M., Tan, B., Le, T. H., Haller, L., Chen, H., et al. 2017. Characterization of metagenomes in urban aquatic compartments reveals high prevalence of clinically relevant antibiotic resistance genes in wastewaters. Frontiers in Microbiology, 8, 2200.
Norouzi, A., Hossieni Nave, H., Mohebi, S., Kandekar Ghareman, M. & taati moghadam, M. 2016. Frequency of plasmid-mediated qnr A, qnr B, and qnr S genes and determination of antibiotic susceptibility among quinolones and fluoroquinolones resistance Escherichia coli isolated from Kerman hospitals. Razi Journal of Medical Sciences, 148, 98-105. (In Persian)
Ohlsen, K., Ternes, T., Werner, G., Wallner, U., Loffler, D., Ziebuhr, W., Witte, W., et al. 2003. Impact of antibiotics on conjugational resistance gene transfer in Staphylococcus aureus in sewage. Environmental Microbiology, 5(8), 711-716.
Ozgumus, O. B., Celik-Sevim, E., Alpay-Karaoglu, S., Sandalli, C. & Sevim, A. 2007. Molecular characterization of antibiotic resistant Escherichia coli strains isolated from tap and spring waters in a coastal region in Turkey. The Journal of Microbiology, 45(5), 379-387.
Pallecchi, L., Riccobono, E., Mantella, A., Bartalesi, F., Sennati, S., Gamboa, H., et al. 2009. High prevalence of qnr genes in commensal Enterobacteria from healthy children in Peru and Bolivia. Antimicrobial Agents and Chemotherapy, 53(6), 2632-2635.
Papa, M., Foladori, P., Guglielmi, L. & Bertanza, G. 2017. How far are we from closing the loop of sewage resource recovery? A real picture of municipal wastewater treatment plants in Italy. Journal of Environmental Management, 198, 9-15
Ramteke, P. W., Gaur, A., Pathak, S. P. & Bhattacharjee, J. W. 1990. Antibiotic resistance of coliforms in drinking water in rural areas. The Indian Journal of Medical Research, 91, 185-188.
Reinthaler, F. F., Posch, J., Feierl, G., Wust, G., Haas, D., Ruckenbauer, G., et al. 2003. Antibiotic resistance of E.coli in sewage and sludge. Water Research, 37(8), 1685-1690.
Robicsek, A., Jacoby, G. A. & Hooper, D. C. 2006. The worldwide emergence of plasmid-mediated quinolone resistance. The Lancet Infectious Diseases, 6(10), 629-640.
Rutgersson, C., Fick, J., Marathe, N., Kristiansson, E., Janzon, A., Angelin, M., et al. 2014. Fluoroquinolones and qnr genes in sediment, water, soil, and human fecal flora in an environment polluted by manufacturing discharges. Environmental Science and Technology, 48(14), 7825-7832.
Salehi, M., Hekmatdoost, M. & Hosseini, F. 2013. Quinolone resistance associated with mexAB-oprM secretory pump from Pseudomonas aeruginosa isolates. Journal of Microbial World, 4(17), 290-298.
Salehi, M., Tabibzadeh, M. & Fallahian, M. R. 2015. Prevalence of qnr gene in clinical Acinetobacter isolates. New Cellular and Molecular Biotechnology Journal, 5(18), 83-88. (In Persian)
Shaghaghi, B., Nakhost Lotfi, M., Mahmmodi, N. S. & Pourshafiei, M. R. 2007. Different strains of enterococci in sewage treatment plants in Tehran. Iranian Journal of Infectious Diseases and Tropical Medicine, 12(37), 61-66. (In Persian)
Silva, J., Castillo, G., Callejas, L., Lopez, H. & Olmos, J. 2006. Frequency of transferable multiple antibiotic resistance amongst coliform bacteria isolated from a treated sewage effluent in Antofagasta, Chile. Electronic Journal of Biotechnology, 9(5), 533-540.
Soleimani Asl, Y., Zibaei, M. & Firoozeh, F. 2013. Detection of qnrA gene among quinolone-resistant Escherichia coli isolated from urinary tract infections in Khorram Abad during 2011-2012. Feyz Journal of Kashan University of Medical Sciences, 17( 5), 488-494.
Strahilevitz, J., Jacoby, G. A., Hooper, D. C. & Robicsek, A. 2009. Plasmid-mediated quinolone resistance: a multifaceted threat. Clinical Microbiology Reviews, 22(4), 664-689.
Tavanania, S., Ranjbar, R. & Sabokbar, A., 2014. Genetic study of antibiotic resistance associated with quinolon antibiotics in Escherichia coli isolates isolated from water resources of Alborz province. Islamic Azad University, Karaj Branch, 35, 36-48. (In Persian)
Vilanova, X., Manero, A., Cerda‐Cuellar, M. & Blanch, A. R. 2004. The composition and persistence of faecal coliforms and enterococcal populations in sewage treatment plants. Journal of Applied Microbiology, 96(2), 279-288.
Wang, M., Guo, Q., Xu X., Wang, X., Ye, X., Wu, S., et al. 2009. New plasmid-mediated quinolone resistance gene, qnr C, found in a clinical isolate of Proteus mirabilis. Antimicrobial Agents and Chemotherapy, 53(5), 1892-1897.
Wayne, p. A. 2012. Performance standards for antimicrobial susceptibility testing. 22th Informational Supplement M100-S22, Institute Clinical and Laboratory Standards, Philadephia, USA.
Xia, R., Ren, Y. & Xu, H. 2013. Identification of plasmid-mediated quinolone resistance qnr genes in multidrug-resistant Gram-negative bacteria from hospital wastewaters and receiving waters in the Jinan area, China. Microbial Drug Resistance, 19(6), 446-456.
Yousefi, S., Mojtahedi, A., Shenagari, M. & Atrkar Roushan, Z. 2015. The relation of qnr genes of the induction of resistance to ciprofloxacin in Escherichia coli. Scientific Journal of Kurdistan University of Medical Sciences, 20(5), 52-60. (In Persian)
Zhang, X. X., Zhang, T. & Fang H. H. 2009. Antibiotic resistance genes in water environment. Applied Microbiology and Biotechnology, 82, 397-414.