Phenotypic and genotypic screening of multi-drug resistant ESBL producing Escherichia coli and Klebsiella pneumoniae from burn wound infection

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Research Paper 01/05/2020
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Phenotypic and genotypic screening of multi-drug resistant ESBL producing Escherichia coli and Klebsiella pneumoniae from burn wound infection

Sohana Al Sanjee, Azizul Haque, Sanchita Banerjee, Md. Shihabul Hoque, Md. Moniruzzaman, Keshob Chandra Das, Md. Ekramul Karim
Int. J. Biosci.16( 5), 145-155, May 2020.
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Abstract

As burn cases are very frequent in Bangladesh, the aim of the present study was to screen the multidrug resistant extended-spectrum-β-lactamase (ESBL) producing Escherichia coli and Klebsiella pneumoniae from burn wound infection. E. coli and K. pneumoniae were isolated on MacConkey agar and Eosin Methylene Blue (EMB) agar and identified by their colony characteristics and biochemical tests. Antimicrobial susceptibility of the isolates was performed by Kirby-Bauer disc diffusion method. After phenotypic ESBL confirmation, isolates were checked for the presence of ESBL genes by polymerase chain reaction (PCR). Among the isolates, K. pneumoniae (62.5%, n = 10) was highly prevalent one followed by E. coli (37.5%, n = 6). E. coli were sensitive to nitrofurantoin (66.67%) while K. pneumoniae were sensitive to amikacin (40%) and gentamicin (40%). Both of the isolates showed complete resistance (100%) to cefotaxime (30 µg), ceftazidime (30 µg) and cefoxitin (30 µg). MAR index was in the range of 0.69-1.0 for all the isolates except, one. All the isolates (100%, n = 16) were phenotypically positive for ESBL production. In PCR analysis, dominant ESBL class was found as blaTEM (87.5%, n = 14) followed by blaSHV (37.5%, n = 6). The co-existence of blaTEM and blaSHV in K. pneumoniae was also observed (50%, n = 5). However, none of the E. coli isolates harbored blaSHV gene. The findings of the present study showed that all the isolates were multidrug resistant and ESBL positive, which might be a serious threat to the clinical management of burn wound infection.

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Abreu AC, Tavares RR, Borges A, Mergulhão F, Simões M. 2013. Current and emergent strategies for disinfection of hospital environments. Journal of Antimicrobial Chemotherapy 68(12), 2718-2732.

Adeyankinnu FA, Motayo BO, Akinduti A, Akinbo J, Ogiogwa JI, Aboderin BW, Agunlejika RA. 2014. A multicenter study of betalactamase resistant Escherichia coli and Klebsiella pneumoniae reveals high level chromosome mediated extended spectrum β-lactamase resistance in Ogun state, Nigeria. Interdisciplinary Perspectives on Infectious Diseases 2014, 819896.

Alam SMS, Kalam MA, Munna MS, Munshi SK, Noor R. 2014. Isolation of pathogenic microorganisms from burn patients admitted in Dhaka Medical College and Hospital and demonstration of their drug resistance traits. Asian Pacific Journal of Tropical Disease 4(5), 402-407.

Bauer AW, Kirby WM, Sherris JC, Turck M. 1966. Antibiotic Susceptibility Testing by a Standardized Single Disk Method. American Journal of Clinical Pathology 45(4), 493-496.

Bayram Y, Parlak M, Aypak C, Bayram I. 2013. Three-year review of bacteriological profile and antibiogram of burn wound isolates in Van, Turkey. International Journal of Medical Sciences 10(1), 19-23.

Bowen-Jones JR, Coovadia YM, Bowen-Jones EJ. 1990. Infection control in a Third World burn facility. Burns 16(6), 445-448.

Bradford PA. 2001. Extended-spectrum beta-lactamases in the 21st century: characterization, epidemiology, and detection of this important resistance threat. Clinical Microbiology Reviews 14, 933-951.

Cappuccino JG, Sherman N. 1996. Microbiology – a laboratory manual. The Benjamin/Cummings Publishing Co. Inc., Menlo Park, California.

Chaudhary U, Aggarwal R. 2004. Extended spectrum β-lactamases (ESBL) – An emerging threat to clinical therapeutics. Indian Journal of Medical Microbiology 22, 75–80.

Chim H, Tan BH, Song C. 2007. Five-year review of infections ina burn intensive care unit: high incidence of Acinetobacter baumannii in a tropical climate. Burns 33(8), 1008-1014.

Church D, Elsayed S, Reid O, Winston B, Lindsay R. 2006. Burn wound infections. Clinical Microbiology Reviews 19(2), 403-434.

CLSI (Clinical and Laboratory Standards Institute). 2011. Performance Standards for Antimicrobial Susceptibility Testing. 21st Informational Supplement M100-S21. Wayne, PA, USA.

CLSI (Clinical and Laboratory Standards Institute). 2017. Performance Standards for Antimicrobial Susceptibility Testing. 27th Edition. Wayne, PA, USA.

Collee JG, Fraser AG, Marmion BP, Simons A. 2015. Mackie & McCartney’s Practical medical microbiology, 8th edition, Churchil Livingstone, USA.

Colom K, Pérez J, Alonso R, Fernández-Aranguiz A, Lariño E, Cisterna R. 2003. Simple and reliable multiplex PCR assay for detection of blaTEM, bla (SHV) and bla OXA-1 genes in Enterobacteriaceae. FEMS Microbiology Letters 223(2), 147–151.

Ekrami A, Kalantar E. 2007. Bacterial infections in burn patients at a burn hospital in Iran. Indian Journal of Medical Research 126, 541-544.

Finch R, FitzGerald J, Howkey P, Teale C, Aymes S, Cookson B. 2012. ESBLs: A Threat to Human and Animal health? Report by the Joint Working Group of DARC and ARHI.  Accessed on 15.04.2020. https://assets.publishing.service.gov.uk/government/uploads/system/uploads/attac ment_data/file/215180/dh_132534.pdf.

Forbes BA, Sahm DF, Weissfeld AS. 1998. Bailley & Scott’s diagnostic microbiology. 10th edition, Mosby Inc., St. Louis Missouri.

Islam MS, Yusuf MA, Chowdhury MSJH, Hossain MA.  2012. ESBL producing Gram negative aerobic bacteria isolated from burn wound infection with their antibiogram in Dhaka. Journal of Science Foundation 10(2), 63-69.

Jacoby GA. 1997. Extended-spectrum beta-lactamases and other enzymes providing resistance to oxyimino-beta-lactams. Infectious Disease Clinics of North America 11, 875–887.

Jones W, Minei J, Barber A. 1990. Bacterial translocation and intestinal atrophy after thermal injury and burn wound sepsis. Annals Surgery 2(11), 399- 405.

Karim MH, Sayad MDA, Yeasmin T. 2017. Molecular identification of TEM and SHV genes in extended spectrum beta-lactamase producing Escherichia coli and Klebsiellae pneumoniae isolates in a tertiary care hospital, Bangladesh. Journal of Pure & Applied Microbiology 11(2), 1189-1198.

Kaur M, Aggarwal A. 2013. Occurrence of the CTX-M, SHV and the TEM genes among the extended spectrum beta-lactamase producing isolates of Enterobacteriaceae in a tertiary care hospital of north India. Journal of Clinical and Diagnostic Research 7, 642-645.

Kavanagh F. 1975. Microbiological diffusion assay II: design and application. Journal of Pharmaceutical Sciences 64, 1224-1229.

Khaleque M, Akter S, Mondal DK, Akhter H, Khan SI, Begum A. 2017. Molecular characterization of extended spectrum β-lactamase producing bacteria isolated from urinary tract infected patients, Bangladesh. Tropical Biomedicine 34(3), 512–523.

Khan ER, Aung MS, Paul SK, Ahmed S, Haque N, Ahamed F, Sarkar SR, Roy S, Rahman MM, Mahmud MC, Hossain MA, Urushibara N, Kawaguchiya M, Sumi A, Kobayashi N. 2018. Prevalence and Molecular Epidemiology of Clinical Isolates of Escherichia coli and Klebsiella pneumoniae Harboring Extended-Spectrum Beta-Lactamase and Carbapenemase Genes in Bangladesh. Microbial Drug Resistance 24, 1568-1579.

Krumpernam PH. 1983. Multiple antibiotic resistance indexing Escherichia coli to identify risk sources of fecal contamination of foods. Applied and Environmental Microbiology 46, 165-170.

Lari AR, Alaghehbandan R. 2000. Nosocomial infections in an Iranian burn care center. Burns 26(8), 737-740.

Livermore DM. 1995. Beta-lactamases in laboratory and clinical resistance. Clinical Microbiology Reviews 8, 557–584.

O’Neil J. 2014. Review on antimicrobial resistance: Tackling a Crisis for the Health and Wealth of Nations. Accessed on 15.04.2020. https://amrreview.org/sites/default/files/AMR%20Review%20Paper%20%20Tackling%20a%20crisis%20for%20the%20health%20and%20wealth%20of%20nations_1.pdf.

Paterson DL, Bonomo RA. 2005. Extended-spectrum beta-lactamases: a clinical update. Clinical Microbiology Reviews 18, 657-686.

Pruitt Jr BA, McManus AT, Kim SH, Goodwin CW. 1998. Burn wound infections: current status. World Journal of Surgery 22(2), 135-145

Rawat D, Nair D. 2010. Extended-spectrum beta-lactamases in Gram Negative Bacteria. Journal of Global Infectious Diseases 2, 263-274.

Roy S, Ahmed MU, Uddin BMM, Ratan ZA, Rajawat M, Mehta V, Zaman SB. 2017. Evaluation of antibiotic susceptibility in wound infections: A pilot study from Bangladesh. F1000 Research 6, 2103.

Santucci SG, Gobara S, Santos CR. 2003. Infections in a burn intensive care unit: experience of seven years. Journal of Hospital Infection 53, 6-13.

Shiju MP, Yashavanth R, Narendra N. 2010. Detection of extended spectrum beta-lactamase production and multidrug resistance in clinical isolates of E. coli and K. pneumoniae in Mangalore. Journal of Clinical and Diagnostic Research 4(3), 2442-2445.

Sultana M, Naeem N, Sultana S, Sultana KF, Mukharjee SK, Hossain MA. 2016. Screening of Extended Spectrum Beta-Lactamase Producing Bacteria in Clinical Liquid Waste. Bangladesh Medical Research Council Bulletin 42, 39-48.

Sultana S, Mawla N, Kawser S, Akhtar N, Ali MK. 2015. Current microbial isolates from wound swab and their susceptibility pattern in a private medical college hospital in Dhaka city. Delta Medical College Journal 3(1), 25-30.

Taneja N, Chari P, Singh M, Singh G, Biswal M, Sharma M. 2013. Evolution of bacterial flora in burn wounds: key role of environmental disinfection in control of infection. International Journal of Burns Trauma 3(2), 102-107.

WHO. 2014. Antimicrobial resistance. In: Global Report on Surveillance. Accessed on 15.04.2020. https://www.who.int/antimicrobialresistance/publications/surveillancereport/en/