Investigation of Phenotypic and mcr-1–Mediated Colistin Resistance in Escherichia coli

This study aimed to investigate colistin resistance in Escherichiacoli isolated from a total of 166 stool samples that were obtainedfrom chickens shipped to slaughter in Turkey and the presenceof the mcr-1 gene, which has been recently discoveredand is one of many genes causing plasmid-mediated colistin resistance.To this end, 200 E. coli isolates were collected and identifiedfrom feces obtained from intestinal samples of the broilerchickens included in this study. To reveal the phenotypic colistinresistance profile, the minimum inhibitory concentration ofcolistin for each isolate was determined using the cation-adjustedbroth microdilution method. Phenotypic colistin resistancewas observed in 15 (7.5%) out of 200 E. coli samples obtainedfrom 7 different poultry slaughterhouses. The presence of themcr-1 gene, which provides plasmid-mediated colistin resistance,was investigated by the polymerase chain reaction methodto detect genotypic colistin resistance. The mcr-1 gene was notdetected in any of the studied samples, including isolates thatexhibited phenotypic colistin resistance. The absence of theplasmid-mediated mcr-1 gene, although colistin resistance wasdetected, suggested that the resistance was of chromosomeorigin or that there were other genes responsible for resistance.However, it is thought that it is necessary to conduct newstudies that will investigate the newly emerging plasmid-mediatedgenes in the future.

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Adiguzel, M. C., Baran, A., Wu, Z., Cengiz, S., Dai, L., Oz, C., Ozmenli, E., Goulart, D. B., & Sahin, O. (2020). Prevalence of colistin resistance in Escherichia coli in Eastern Turkey and genomic characterization of an mcr-1 positive strain from retail chicken meat. Microbial Drug Resistance, doi.org/10.1089/mdr.2020.0209. [Crossref]

Anjum, M. F., Duggett, N. A., AbuOun, M., Randall, L., Nunez-Garcia, J., Ellis, R. J., Horton, R., Brena, C., Williamson, S., Martelli, S., Davies, R., & Teale, C. (2016). Colistin resistance in Salmonella and Escherichia coli isolates from a pig farm in Great Britain. The Journal of Antimicrobial Chemotherapy, 71(8), 2306-2313. [Crossref]

Aghapour, Z., Hasani, A., Aghazadeh, M., & Ahangarzadeh, M. (2019). Genes involved in colistin resistance of gram-negative isolates in the northwest of Iran. Gene Reports, 14, 81-86. [Crossref]

Baquero, F., & Martínez, J. L. (2000). Mutation frequencies and antibiotic resistance. Antimicrobial Agents and Chemotherapy, 44(7), 1771- 1777. [Crossref]

Borowiak, M., Fischer, J., Hammerl, J. A., Hendriksen, R. S., Szabo, I., & Malorny, B. (2017). Identification of a novel transposon-associated phosphoethanolamine transferase gene, mcr-5, conferring colistin resistance in d-tartrate fermenting Salmonella enterica subsp. enterica serovar Paratyphi B. Journal of Antimicrobial Chemotherapy, 72(12), 3317-3324. [Crossref]

Caniaux, I., Belkum, A. Van, Zambardi, G., Poirel, L., & Gros, M. F. (2016). MCR : modern colistin resistance. European Journal of Clinical Microbiology & Infectious Diseases, 36(3), 415-420. [Crossref]

Carattoli, A., Villa L., Feudi, C., Curcio, L., Orsini, S., Luppi, A., Pezzotti, G., & Magistrali, C. F. (2017). Novel plasmid-mediated colistin resistance mcr-4 gene in Salmonella and Escherichia coli, Italy 2013, Spain and Belgium, 2015 to 2016. Euro Surveillance, 22(31), 30589. [Crossref]

Carroll, L. M., Gaballa, A., Guldimann, C., Sullivan, G., Henderson, L. O., & Wiedmann, M. (2019). Identification of novel mobilized colistin resistance gene mcr-9 in a multidrug-resistant, colistin-susceptible salmonella enterica serotype typhimurium isolate. mBio, 10(3), e00853-19. [Crossref]

Catry, B., Cavaleri, M., Baptiste, K., Grave, K., Grein, K., Holm, A., Jukes, H., Liebana, E., Lopez Navas, A., Mackay, D., Magiorakos, A.P., Moreno Romo, M. A., Moulin, G., Muñoz Madero, C., Matias Ferreira Pomba, M. C., Powell, M., Pyörälä, S., Rantala, M., Ružauskas, M., Sanders, P., Teale, C., Threlfall, E. J., Törneke, K., van Duijkeren, E., & Edo, J. T. (2015). Use of colistin-containing products within the European Union and European Economic Area (EU/EEA): development of resistance in animals and possible impact on human and animal health. International Journal of Antimicrobial Agents, 46(3), 297- 306. [Crossref]

Cavaco, L., & Hendriksen, R. (2015). PCR for plasmid-mediated colistin resistance gene mcr-1, protocol optimized at Statens Serum Institut, Copenhagen, Denmark: Available from: https://www.eurl-ar.eu/ (Accessed on: December, 2015).

Di Pilato, V., Arena, F., Tascini, C., Cannatelli, A., Henrici De Angelis, L., Fortunato, S., Giani, T., Menichetti, F., & Rossolini, G.M. (2016). mcr- 1.2: a new mcr variant carried on a transferable plasmid from a colistin-resistant KPC carbapenemase-producing Klebsiella pneumoniae strain of sequence type 512. Antimicrobial Agents and Chemotherapy, 60(9), 5612-5615. [Crossref]

Etebu, E., & Ukpong, M. (2016). Bacterial resistance to antibiotics: Update on molecular perspectives. Microbiology Research International, 4(4), 40-49.

EUCAST Recommendations for MIC determination of colistin (Polymyxin E) As recommended by joint CLSI-EUCAST Polymyxin Breakpoints Working Group: Available from: www.eucast.org. (Accessed on: March, 22nd, 2016)

Falgenhauer, L., Waezsada, S. E., Yao, Y., Imirzalioglu, C., Käsbohrer, A., Roesler, U., Michael, G.B., Schwarz, S., Werner, G., Kreienbrock, L., & Chakraborty, T. (2016). Colistin resistance gene mcr-1 in extended- spectrum β-lactamase-producing and carbapenemase-producing gram-negative bacteria in Germany. The Lancet Infectious Diseases, 16(3), 282-283. [Crossref]

Garcia-Graells, C., De Keersmaecker, S., Vaneste, K., Pochet, B., Vermeersch, K., Roosens, N., Dierick, K., & Botteldoorn, N. (2018). Detection of plasmid-mediated colistin resistance, mcr-1 and mcr-2 genes, in Salmonella spp. isolated from food at retail in Belgium from 2012 to 2015. Foodborn Pathogens and Disease, 15(2), 114- 117. [Crossref]

Grami, R., Mansour, W., Mehri, W., Bouallègue, O., Boujaâfar, N., Madec, J., & Haenni, M. (2016). Impact of food animal trade on the spread of mcr-1-mediated colistin resistance, Tunisia, July 2015. Eurosurveillance, 21(8), 1-5. [Crossref]

Haenni, M., Poirel, L., Kieffer, N., Châtre, P., Saras, E., Métayer, V., Dumoulin, R., Nordmann, P., & Madec, J. Y. (2016). Co-occurrence of extended spectrum β lactamase and mcr-1 encoding genes on plasmids. The Lancet Infectious Diseases, 16(3), 281-282. [Crossref]

Hasman, H., Hammerum, A. M., Hansen, F., Hendriksen, R. S., Olesen, B., Agersø, Y., Zankari, E., Leekitcharoenphon, P., Stegger, M., Kaas, R. S., Cavaco, L. M., Hansen, D. S., Aarestrup, F. M., & Skov, R. L. (2015). Detection of mcr-1 encoding plasmid-mediated colistin-resistant Escherichia coli isolates from human bloodstream infection and imported chicken meat, Denmark 2015. Eurosurveillance, 20(49), 1-5. [Crossref]

International Organization for Standardization (ISO) (2006). Clinical laboratory testing and in vitro diagnostic test systems: Susceptibility testing of infectious agents and evaluation of performance of antimicrobial susceptibility test devices. ISO 20776-1.2006; 1-19. Irrgang, A., Roschanski, N., Tenhagen, B. A., Grobbel, M., Skladnikiewicz- Ziemer, T., Thomas, K., Roesler, U., & Käsbohrer, A. (2016). Prevalence of mcr-1 in E. coli from Livestock and Food in Germany, 2010–2015. Plos One, 11, e0159863. [Crossref]

Kempf, I., Fleury, M. A., Drider, D., Bruneau, M., Sanders, P., Chauvin, C., Madec, J. Y., & Jouy, E. (2013). What do we know about resistance to colistin in Enterobacteriaceae in avian and pig production in Europe? International Journal of Antimicrobial Agents, 42(5), 379-383. [Crossref]

Kuo, S. C., Huang, W. C., Wang, H. Y., Shiau, Y. R., Cheng, M. F., & Lauderdale, T. L. (2016). Colistin resistance gene mcr -1 in Escherichia coli isolates from humans and retail meats, Taiwan. Journal of Antimicrobial Chemotherapy, 71(8), 2327-2329. [Crossref]

Kurekci, C., Aydin, M., Nalbantoglu, O. U., & Gundogdu, A. (2018). First report of Escherichia coli carrying the mobile colistin resistance gene mcr-1 in Turkey. Journal of Global Antimicrobial Resistance, 15, 169-170. [Crossref]

Kusumoto, M., Ogura, Y., Gotoh, Y., Iwata, T., Hayashi, T., & Akiba, M. (2016). Colistin-resistant mcr-1 positive pathogenic Escherichia coli in swine. Emerging Infectious Diseases, 22(7), 2014-2016. [Crossref]

Liu, Y. Y., Wang, Y., Walsh, T. R., Yi, L. X., Zhang, R., Spencer, J., Doi, Y., Tian, G., Dong, B., Huang, X., Yu, L. F., Gu, D., Ren, H., Chen, X., Lv, L., He, D., Zhou, H., Liang, Z., Liu, J. H., & Shen, J. (2016). Emergence of plasmid- mediated colistin resistance mechanism mcr-1 in animals and human beings in China: a microbiological and molecular biological study. The Lancet Infectious Diseases, 16(2), 161-168. [Crossref]

McGann, P., Snesrud, E., Maybank, R., Corey, B., Ong, A. C., Clifford, R., Hinkle, M., Whitman, T., Lesho, E., & Schaecher, K. E. (2016). Escherichia coli Harboring mcr-1 and blaCTX-M on a Novel IncF Plasmid: first report of mcr-1 in the USA. Antimicrobial Agents Chemotherapy, 60(7), 4420-4421. [Crossref]

Ovejero, C. M., & Muniesa, M. (2017). Spread of mcr-1 -carrying Enterobacteriaceae in sewage water from Spain. Journal of Antimicrobial Chemotherapy, 72(4), 1050-1053. [Crossref]

Principe, L., Piazza A., Mauri, C., Anesi A., Bracco, S., Brigante, G., Casari, E., Agrappi, C., Caltagirone, M., Novazzi, F., Migliavacca, R., Pagani, L., & Luzzaro, F. (2018). Multicenter prospective study on the prevalence of colistin resistance in Escherichia coli: relevance of mcr-1-positive clinical isolates in Lombardy, Northern Italy. Infection and Drug Resistance, 11, 377-385. [Crossref]

Sekyere, O. J., Govinden, U., Bester, L. A., & Essack, S. Y. (2016). Colistin and tigecycline resistance in carbapenemase-producing gram negative bacteria: emerging resistance mechanisms and detection methods. Journal of Applied Microbiology, 121(3), 601-617. [Crossref]

Skov, R. L., & Monnet, D. L. (2016). Plasmid-mediated colistin resistance (mcr-1 gene): three months later, the story unfolds. Eurosurveillance, 21(9), 30155. [Crossref]

Stoesser, N., Mathers, A. J., Moore, C. E., Day, N. P. J., & Crook, D. W. (2016). Colistin resistance gene mcr-1 and pHNSHP45 plasmid in human isolates of Escherichia coli and Klebsiella pneumoniae. The Lancet Infectious Diseases, 16(3), 285-286. [Crossref]

Suzuki, S., Ohnishi, M., Kawanishi, M., Akiba, M., & Kuroda, M. (2016). Investigation of a plasmid genome database for colistin-resistance gene mcr-1. The Lancet Infectious Diseases, 16(3), 284-285. [Crossref]

Tenover, F. C. (2006). Mechanisms of antimicrobial resistance in bacteria. American Journal of Infection Control, 19(6), 3-10. [Crossref]

Wang, C., Feng, Yu., Liu, L., Wei, Li., Kang, M., & Zong, Z. (2020). Identification of novel mobile colistin resistance gene mcr-10. Emerging Microbes and Infections, 9(1), 508-516. [Crossref]

Wang, X., Wang, Y., Zhou, Y., Li, J., Yin, W., Wang, S., Zhang, S., Shen, J., Shen, Z., & Wang, Y. (2018). Emergence of a novel mobile colistin resistance gene, mcr-8, in NDM-producing Klebsiella pneumoniae article. Emerging Microbes and Infections, 7(1), 1-9. [Crossref]

Watkins, R. R. (2016). Overview: Global and local impact of antibiotic resistance antibiotic resistance infections agriculture public health. Infectious Disease Clinics of North America, 30(2), 313-322. [Crossref]

Watkins, R. R., Smith, T. C., & Bonomo, R. A. (2016). On the path to untreatable infections: colistin use in agriculture and the end of “last resort” antibiotics. Expert Review of Anti-infective Therapy, 14(9), 785-788. [Crossref]

WHO (World Health Organization) (2016). Available from: http://www.who.int (Accessed in February 2019).

Xavier, B. B., Lammens, C., Ruhal, R., Kumar-Singh, S., Butaye, P., Goossens, H., & Malhotra-Kumar, S. (2016a). Identification of a novel plasmid-mediated colistin-resistance gene, mcr-2, in Escherichia coli, Belgium, June 2016. Eurosurveillance, 21(27), 611. [Crossref]

Xavier, B. B., Lammens, C., Butaye, P., Goossens, H., & Malhotra-Kumar, S. (2016b). Complete sequence of an IncFII plasmid harbouring the colistin resistance gene mcr-1 isolated from Belgian pig farms. Journal of Antimicrobial Chemotherapy, 71(8), 2342-2344. [Crossref]

Yang, Y. Q., Li, Y. X., Song, T., Yang, Y. X., Jiang, W., Zhang, A. Y., Guo, X. Y., Liu, B. H., Wang, Y. X., Lei, C. W., Xiang, R., & Wang, H. N. (2017). Colistin resistance gene mcr-1 and its variant in Escherichia coli isolates from chickens in China. Antimicrobial Agents and Chemotherapy, 61(5), e01204-e01216. [Crossref]

Yang, Y. Q, Li, Y. X., Lei, C. W., Zhang A. Y., & Wang, H.N.(2018). Novel plasmid- mediated colistin resistance gene mcr-7.1 in Klebsiella pneumoniae. Journal Antimicrob Chemotherapy, 73(7), 1791-1795. [Crossref]

Yin, W., Li, H., Shen, Y., Liu, Z., Wang, S., Shen, Z., Zhang, R., Walsh, T.R., Shen, J., & Wang, Y. (2017). Novel plasmid-mediated colistin resistance gene mcr-3 in Escherichia coli. American Society For Microbiology, 8(3), e00543. [Crossref]

Zurfuh, K., Poirel, L., Nordmann, P., Nüesch-Inderbinen, M., Hächler, H., & Stephan, R. (2016). Occurrence of the plasmid-borne mcr-1 colistin resistance gene in extended-spectrum-lactamase-producing enterobacteriaceae in river water and imported vegetable samples in Switzerland. Antimicrobial Agents and Chemotherapy, 60(4), 2594-2595. [Crossref]