Colistin has been regarded as the last line antibiotic for treatment of infections caused by multidrug resistant gram-negative bacteria. Therefore, the increasing emergence of colistin resistance among gram-negative bacteria represents a serious problem. The objective of this study was to characterize the effectiveness of the chemically synthesized thanatin in linear form against colistin-resistant E. coli isolated from a pig farm in China. Agar diffusion assay and broth microdilution test were employed to analyze the susceptibility of colistin-sensitive E. coli (ATCC25922) and colistin-resistant E. coli (SHP45) to linear thanatin (L-thanatin). Combinatory effect of linear thanatin and colistin against E. coli was also determined by fractional inhibition concentration index (FICI) analysis. The results showed that L-thanatin at a concentration of 1 mg/ml produced larger inhibition zone on agar against ATCC25922 than SHP45. In the quantitative microdilution test, L-thanatin had the same MIC of 3.2 μg/ml for ATCC25922 and SHP45. Based on the FICI analysis, additive effect was obtained with 1.56 μg/ml of L-thanatin and 0.125 μg/ml of colistin for ATCC25922; but with 1.56 μg/ml of L-thanatin and 0.25 μg/ml of colistin or with 2 μg/ml of colistin and 0.39 μg/ml of L-thanatin for SHP45. These data proved that L-thanatin is an effective antimicrobial peptide against colistin-resistant E. coli .
Keywords<i>E. coli</i>Colistin ResistanceLinear ThanatinMinimum Inhibition ConcentrationFractional Inhibition Concentration Index
Thabit, A.K., Crandon, J.L. and Nicolau, D.P. (2015) Antimicrobial Resistance: Impact on Clinical and Economic Outcomes and the Need for New Antimicrobials. Expert Opinion on Pharmacotherapy, 16, 159-177. https://doi.org/10.1517/14656566.2015.993381
Moxon, C.A. and Paulus, S. (2016) Beta-Lactamases in Enterobacteriaceae Infections in Children. Journal of Infection, 72, S41-S49. https://doi.org/10.1016/j.jinf.2016.04.021
Ainsworth, G.C., Brown, A.M. and Brownlee, G. (1947) Aerosporin, an Antibiotic Produced by Bacillus aerosporus Greer. Nature, 159, 263.
Schoenbach, E.B. and Bryer, M.J. (1948) The Clinical Use of Polymyxin. Bulletin of Johns Hopkins Hospital, 82, 637-639.
Brown, P. and Dawson, M.J. (2017) Development of New Polymyxin Derivatives for Multi-Drug Resistant Gram-Negative Infections. The Journal of Antibiotics (Tokyo), 70, 386-394. https://doi.org/10.1038/ja.2016.146
Rhouma, M., Beaudry, F., Thériault, W. and Letellier, A. (2016) Colistin in Pig Production: Chemistry, Mechanism of Antibacterial Action, Microbial Resistance Emergence, and One Health Perspectives. Frontiers in Microbiology, 7, 1789.
Harada, K., Asai, T., Kojima, A., Oda, C., Ishihara, K. and Takahashi, T. (2005) Antimicrobial Susceptibility of Pathogenic Escherichia coli Isolated from Sickcattle and Pigs in Japan. Journal of Veterinary Medical Science, 67, 999-1003.
Enne, V.I., Cassar, C., Sprigings, K., Woodward, M.J. and Bennett, P.M. (2008) A High Prevalence of Antimicrobial Resistant Escherichia coli Isolated from Pigs and a Low Prevalence of Antimicrobial Resistant E. coli from Cattle and Sheep in Great Britain at Slaughter. FEMS Microbiology Letters, 278, 193-199. https://doi.org/10.1111/j.1574-6968.2007.00991.x
Lu, L., Dai, L., Wang, Y., Wu, C., Chen, X., Li, L., Qi, Y., Xia, L. and Shen, J. (2010) Characterization of Antimicrobial Resistance and Integrons among Escherichia coli Isolated from Animal Farms in Eastern China. Acta Tropica, 113, 20-25. https://doi.org/10.1016/j.actatropica.2009.08.028
Olaitan, A.O., Morand, S. and Rolain, J.M. (2014) Mechanisms of Polymyxin Resistance: Acquired and Intrinsic Resistance in Bacteria. Frontiers in Microbiology, 5, 643.
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. and 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, 161-168. https://doi.org/10.1016/S1473-3099(15)00424-7
Olaitan, A.O., Thongmalayvong, B., Akkhavong, K., Somphavong, S., Paboriboune, P., Khounsy, S., Morand, S. and Rolain, J.M. (2015) Clonal Transmission of a Colistin-Resistant Escherichia coli from a Domesticated Pig to a Human in Laos. Journal of Antimicrobial Chemotherapy, 70, 3402-3404.
Olaitan, A.O., Chabou, S., Okdah, L., Morand, S. and Rolain, J.M. (2016) Dissemination of the MCR-1 Colistin Resistance Gene. The Lancet Infectious Diseases, 16, 147. https://doi.org/10.1016/S1473-3099(15)00540-X
Fehlbaum, P., Bulet, P., Chemysh, S., Briand, J.P., Rousse, J.P., Letellier, L., et al. (1996) Structure Activity Analysis of Thanatin, a 21-Residue Inducible Insect Defense Peptide with Sequence Homology to Frog Skin Antimicrobial Peptides. Proceedings of the National Academy of Sciences, 93, 1221-1225. https://doi.org/10.1073/pnas.93.3.1221
Mandard, N., Sodano, P., Labbe, H., Bonmatin, J.M., Bulet, P., Hetru, C., Ptak, M. and Vovelle, F. (1998) Solution Structure of Thanatin, a Potent Bactericidal and Fungicidal Insect Peptide, Determined from Proton Two-Dimensional Nuclear Magnetic Resonance Data. European Journal of Biochemistry, 256, 404-410. https://doi.org/10.1046/j.1432-1327.1998.2560404.x
Pagès, J.M., Dimarcq, J.L., Quenin, S. and Hetru, C. (2003) Thanatin Activity on Multidrug Resistant Clinical Isolates of Enterobacter aerogenes and Klebsiella pneumoniae. International Journal of Antimicrobial Agents, 22, 265-269. https://doi.org/10.1016/S0924-8579(03)00201-2
Wu, G., Fan, X., Li, L., Wang, H., Ding, J., Hongbin, W., Zhao, R., Gou, L., Shen, Z. and Xi, T. (2010) Interaction of Antimicrobial Peptide s-Thanatin with Lipopolysaccharide in Vitro and in an Experimental Mouse Model of Septic Shock Caused by a Multidrug-Resistant Clinical Isolate of Escherichia coli. International Journal of Antimicrobial Agents, 35, 250-254. https://doi.org/10.1016/j.ijantimicag.2009.11.009
Wu, G., Li, X., Fan, X., Wu, H., Wang, S., Shen, Z. and Xi, T. (2011) The Activity of Antimicrobial Peptide S-Thanatin Is Independent on Multidrug-Resistant Spectrum of Bacteria. Peptides, 32, 1139-1145. https://doi.org/10.1016/j.peptides.2011.03.019
Edwards, I.A., Elliott, A.G., Kavanagh, A.M., Zuegg, J., Blaskovich, M.A. and Cooper, M.A. (2016) Contribution of Amphipathicity and Hydrophobicity to the Antimicrobial Activity and Cytotoxicity of β-Hairpin Peptides. ACS Infectious Diseases, 2, 442-450. https://doi.org/10.1021/acsinfecdis.6b00045
Ma, B., Niu, C., Zhou, Y., Xue, X., Meng, J., Luo, X. and Hou, Z. (2016) The Disulfide Bond of the Peptide Thanatin Is Dispensible for Its Antimicrobial Activity in Vivo and in Vitro. Antimicrobial Agents Chemotherapy, 60, 4283-4289. https://doi.org/10.1128/AAC.00041-16
Boulanger, N., Munks, R.J., Hamilton, J.V., Vovelle, F., Brun, R., Lehane, M.J. and Bulet, P. (2002) Epithelial Innate Immunity. A Novel Antimicrobial Peptide with Antiparasitic Activity in the Blood-Sucking Insect Stomoxys calcitrans. Journal of Biological Chemistry, 277, 49921-49926. https://doi.org/10.1074/jbc.M206296200
Del Valle, P., Garcia-Armesto, M.R., De Arriaga, D., Alez-Donquiles, C.G., Rodriguez-Fernandez, P. and Rua, J. (2016) Antimicrobial Activity of Kaempferol and Resveratrol in Binary Combinations with Parabens or Propyl Gallate against Enterococcus faecalis. Food Control, 61, 213-220. https://doi.org/10.1016/j.foodcont.2015.10.001
Lee, Y.S., Kang, O.H., Choi, J.G., Oh, Y.C., Chae, H.S., Kim, J.H., Park, H., Sohn, D.H., Wang, Z.T. and Kwon, D.Y. (2008) Synergistic Effects of the Combination of Galangin with Gentamicin against Methicillin-Resistant Staphylococcus aureus. The Journal of Microbiology, 46, 283-288. https://doi.org/10.1007/s12275-008-0012-7
Alonso, C.A., Zarazaga, M., Ben Sallem, R., Jouini, A., Ben Slama, K. and Torres, C. (2017) Antibiotic Resistance in Escherichia coli in Husbandry Animals: The African Perspective. Letters in Applied Microbiology, 64, 318-334. https://doi.org/10.1111/lam.12724
Zavascki, A.P., Goldani, L.Z., Li, J. and Nation, R.L. (2007) Polymyxin B for the Treatment of Multidrug-Resistant Pathogens: A Critical Review. Journal of Antimicrobial Chemotherapy, 60, 1206-1215. https://doi.org/10.1093/jac/dkm357
Bollenbach, T. (2015) Antimicrobial Interactions: Mechanisms and Implications for Drug Discovery and Resistance Evolution. Current Opinion in Microbiology, 27, 1-9. https://doi.org/10.1016/j.mib.2015.05.008
Mohammadi, M., Khayat, H., Sayehmiri, K., Soroush, S., Sayehmiri, F., Delfani, S., Bogdanovic, L. and Taherikalani, M. (2017) Synergistic Effect of Colistin and Rifampin against Multidrug Resistant Acinetobacter baumannii: A Systematic Review and Meta-Analysis. The Open Microbiology Journal, 11, 63-71. https://doi.org/10.2174/1874285801711010063
Lenhard, J.R., Nation, R.L. and Tsuji, B.T. (2016) Synergistic Combinations of Polymyxins. International Journal of Antimicrobial Agents, 48, 607-613. https://doi.org/10.1016/j.ijantimicag.2016.09.014
Velkov, T., Thompson, P.E., Nation, R.L. and Li, J. (2010) Structure-Activity Relationships of Polymyxin Antibiotics. Journal of Medicinal Chemistry, 53, 1898-1916. https://doi.org/10.1021/jm900999h