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Abstract
Antibiotic-resistant bacteria represent a global risk to public health. Horizontal gene transfer, a common mechanism for genetic exchange in bacteria, plays an essential role in the acquisition of resistance genes. In this work, we evaluated the effect of sub-lethal concentrations of antibiotics on plasmid transfer by conjugation and transformation in the opportunistic pathogen Klebsiella pneumoniae. Despite not being naturally competent, this bacterium could acquire extracellular DNA from various plasmids at a very low frequency, which increased upon incubating cells with the aminoglycoside antibiotics amikacin and gentamicin. Transfer by conjugation analyzed using a clinical isolate carrying plasmid pNDM-1 also increased in the presence of sub-lethal concentrations of antibiotics. An RNAseq analysis showed differential expression of several genes when cells were incubated in the presence of sub-lethal concentrations of amikacin suggesting metabolic and regulatory changes, as well as alteration of cell envelope components that could affect the uptake of foreign DNA. These results suggest that sub-lethal concentrations of some aminoglycosides, in particular amikacin, can promote the transfer of resistance-bearing genetic elements in K. pneumoniae, which is relevant for understanding the spread of resistance determinants in this human pathogen.
References
Allen, H. K., Donato, J., Wang, H. H., Cloud-Hansen, K. A., Davies, J., Handelsman, J. (2010). Call of the wild: Antibiotic resistance genes in natural environments. Nat Rev Microbiol. 8 (4): 251-259.
Allen, H. K., Moe, L. A., Rodbumrer, J., Gaarder, A., Handelsman, J. (2009). Functional metagenomics reveals diverse β-lactamases in a remote Alaskan soil. ISME J. 3 (2): 243-251.
Anders, S. & Huber, W. (2010). Differential expression analysis for sequence count data. Genome Biol. 11 (10): R106.
Andersson, D. I. & Hughes, D. (2014). Microbiological effects of sublethal levels of antibiotics. Nat Rev Microbiol. 12 (7): 465-478.
Andrews, J. M. (2002). Determination of minimum inhibitory concentrations. J Antimicrob Chemother. 49 (6): 1049-1049.
Baharoglu, Z., Krin, E., Mazel, D. (2013). RpoS Plays a Central Role in the SOS Induction by Sub-Lethal Aminoglycoside Concentrations in Vibrio cholerae. PLoS Genet. 9 (4): e1003421.
Balestrino, D., Ghigo, J.-M., Charbonnel, N., Haagensen, J. A. J., Forestier, C. (2008). The characterization of functions involved in the establishment and maturation of Klebsiella pneumoniae in vitro biofilm reveals dual roles for surface exopolysaccharides. Environ Microbiol. 10 (3): 685-701.
Barr, V., Barr, K., Millar, M. R., Lacey, R. W. (1986). β-Lactam antibiotics increase the frequency of plasmid transfer in Staphylococcus aureus. J Antimicrob Chemother. 17 (4): 409-413.
Barrero, L. I., Castillo, J. S., Leal, A. L., Sánchez, R., Cortés, J. A., Álvarez, C. A., González, A. L. (2014). Impacto económico de la resistencia a meticilina en pacientes con bacteriemia por Staphylococcus aureus en hospitales de Bogotá. Biomédica. 34 (3): 345-353.
Benjamini, Y. & Hochberg, Y. (1995). Controlling the False Discovery Rate: A Practical and Powerful Approach to Multiple Testing. J R Stat Soc Ser B. 57: 289-300.
Bhullar, K., Waglechner, N., Pawlowski, A., Koteva, K., Banks, E. D., Johnston, M. D., … Wright, G. D. (2012). Antibiotic Resistance Is Prevalent in an Isolated Cave Microbiome. PLoS One. 7 (4): e34953.
Bloemberg, G. V, Wijfjes, A. H., Lamers, G. E., Stuurman, N., Lugtenberg, B. J. (2000). Simultaneous imaging of Pseudomonas fluorescens WCS365 populations expressing three different autofluorescent proteins in the rhizosphere: New perspectives for studying microbial communities. Mol Plant Microbe Interact. 13 (11): 1170-1176.
Blokesch, M. (2016). Natural competence for transformation. Curr Biol. 26 (21): R1126-R1130.
Bolger, A. M., Lohse, M., Usadel, B. (2014). Trimmomatic: A flexible trimmer for Illumina sequence data. Bioinformatics. 30 (15): 2114-2120.
Carlet, J., Jarlier, V., Harbarth, S., Voss, A., Goossens, H., Pittet, D., Participants of the 3rd World Healthcare-Associated Infections Forum. (2012). Ready for a world without antibiotics? The Pensières Antibiotic Resistance Call to Action. Antimicrob Resist Infect Control. 1 (1): 11.
Cienfuegos-Gallet, A. V, Ocampo-de Los Ríos, A. M., Sierra-Viana, P., Ramírez-Brinez, F., Restrepo-Castro, C., Roncancio-Villamil, G., … Jiménez, J. N. (2019). Risk factors and survival of patients infected with carbapenem-resistant Klebsiella pneumoniae in a KPC endemic setting: a case-control and cohort study. BMC Infect Dis. 19 (1): 830.
Clinical & Laboratory Standards Institute - CLSI. (2015). Performance standards for antimicrobial susceptibility testing: 25th informational supplement. In P. A. Wayne (Ed.), CLSI document M100-S25.
Cosgrove, S. E. (2006). The Relationship between Antimicrobial Resistance and Patient Outcomes: Mortality, Length of Hospital Stay, and Health Care Costs. Clin Infect Di. 42 (Supplement_2): S82–S89.
Cuzon, G., Naas, T., Correa, A., Quinn, J. P., Villegas, M.-V., Nordmann, P. (2013). Dissemination of the KPC-2 carbapenemase in non-Klebsiella pneumoniae enterobacterial isolates from Colombia. Int J Antimicrob Agents. 42 (1): 59-62.
D’Costa, V. M., King, C. E., Kalan, L., Morar, M., Sung, W. W. L., Schwarz, C., … Wright, G. D. (2011). Antibiotic resistance is ancient. Nature. 477 (7365): 457-461.
Dalhoff, A. A. (2018). Membrane Interactions of Antibacterial Agents. Trends Clin Microbiol. 1 (1): 04–48.
Escobar-Pérez, J. A., Olarte-Escobar, N. M., Castro-Cardozo, B., Valderrama-Márquez, I. A., Garzón-Aguilar, M. I., Martínez-de la Barrera, L., … Vanegas-Gómez, N. (2013). Outbreak of NDM-1-producing Klebsiella pneumoniae in a neonatal unit in Colombia. Antimicrob Agents Chemother. 57 (4): 1957-1960.
Finkel, S. E. & Kolter, R. (2001). DNA as a nutrient: Novel role for bacterial competence gene homologs. J Bacteriol. 183 (21): 6288-6293.
Frost, L. S. & Koraimann, G. (2010). Regulation of bacterial conjugation: Balancing opportunity with adversity. Future Microbiol. 5 (7): 1057-1071.
Gaze, W. H., Krone, S. M., Larsson, D. G. J., Li, X.-Z., Robinson, J. A., Simonet, P., Zhu, Y.-G. (2013). Influence of Humans on Evolution and Mobilization of Environmental Antibiotic Resistome. Emerg Infect Dis. 19 (7): e120871.
Harrison, J. J., Stremick, C. a, Turner, R. J., Allan, N. D., Olson, M. E., Ceri, H. (2010). Microtiter susceptibility testing of microbes growing on peg lids: A miniaturized biofilm model for high-throughput screening. Nat Protoc. 5 (7): 1236-1254.
Hawser, S. P., Bouchillon, S. K., Hoban, D. J., Hackel, M., Johnson, J. L., Badal, R. E. (2009). Klebsiella pneumoniae isolates possessing KPC β-lactamase in Israel, Puerto Rico, Colombia and Greece. Int J Antimicrob Agents. 34 (4): 384-385.
Hoffman, L. R., D’Argenio, D. A., MacCoss, M. J., Zhang, Z., Jones, R. A., Miller, S. I. (2005). Aminoglycoside antibiotics induce bacterial biofilm formation. Nature. 436 (7054): 1171-1175.
Holt-Harris, J. E. & Teague, O. (1916). A new culture medium for the isolation of Bacillus typhosus from stools. J Infect Dis. 18 (6): 596-600.
Huertas, M. G., Zárate, L., Acosta, I. C., Posada, L., Cruz, D. P., Lozano, M., Zambrano, M. M. (2014). Klebsiella pneumoniae yfiRNB operon affects biofilm formation, polysaccharide production and drug susceptibility. Microbiology. 160: 2595-2606.
Instituto Nacional de Salud (INS). Resultados del Programa de Vigilancia por Laboratorio de Resistencia antimicrobiana en Infecciones Asociadas a la Atención en Salud (IAAS) 2016. Accessed on Nov, 2019. https://www.ins.gov.co/buscador-eventos/Informacin%20de%20laboratorioInforme%20Vigilancia%20por%20Laboratorio%20Resistencia%20Antimicrobiana%20y%20Whonet%20IAAS%202016.pdf
Jacoby, G. A. & Han, P. (1996). Detection of extended-spectrum beta-lactamases in clinical isolates of Klebsiella pneumoniae and Escherichia coli. J Clin Microbiol. 34 (4): 908-911.
Jones, R. N., Guzmán-Blanco, M., Gales, A. C., Gallegos, B., Castro, A. L. L., Martino, M. D. V., … Castanheira, M. (2013). Susceptibility rates in Latin American nations: Report from a regional resistance surveillance program (2011). Brazilian J Infect Dis. 17 (6): 672-681.
Jutkina, J, Marathe, N. P., Flach, C.-F., Larsson, D. G. J. (2018). Antibiotics and common antibacterial biocides stimulate horizontal transfer of resistance at low concentrations. Sci Total Environ. 616-617: 172-178.
Jutkina, J., Rutgersson, C., Flach, C.-F., Joakim-Larsson, D. G. (2016). An assay for determining minimal concentrations of antibiotics that drive horizontal transfer of resistance. Sci Total Environ. 548-549: 131-138.
Kado, C. I. & Liu, S. T. (1981). Rapid procedure for detection and isolation of large and small plasmids. J Bacteriol. 145 (3): 1365-1373.
Kopylova, E., Noé, L., Touzet, H. (2012). SortMeRNA: Fast and accurate filtering of ribosomal RNAs in metatranscriptomic data. Bioinformatics. 28 (24): 3211-3217.
Kumarasamy, K. K., Toleman, M. A., Walsh, T. R., Bagaria, J., Butt, F., Balakrishnan, R., … Woodford, N. (2010). Emergence of a new antibiotic resistance mechanism in India, Pakistan, and the UK: A molecular, biological, and epidemiological study. Lancet Infect Dis. 10 (9): 597-602.
Landecker, H. (2016). Antibiotic Resistance and the Biology of History. Body Soc. 22 (4): 19-52. Leal, A.L. (2016). Boletín GREBO, Número 9, Bogotá, 2016, ISSN 2027- 0860. Bogotá.
Lee, C.-R., Lee, J. H., Park, K. S., Jeon, J. H., Kim, Y. B., Cha, C.-J., … Lee, S. H. (2017). Antimicrobial Resistance of Hypervirulent Klebsiella pneumoniae: Epidemiology, Hypervirulence-Associated Determinants, and Resistance Mechanisms. Front Cell Infect Microbiol. 7: 483.
Lin, J. T., Connelly, M. B., Amolo, C., Otani, S., Yaver, D. S. (2005). Global transcriptional response of Bacillus subtilis to treatment with subinhibitory concentrations of antibiotics that inhibit protein synthesis. Antimicrob Agents Chemother. 49 (5): 1915-1926.
Liu, G., Bogaj, K., Bortolaia, V., Olsen, J. E., Thomsen, L. E. (2019). Antibiotic-Induced, Increased Conjugative Transfer Is Common to Diverse Naturally Occurring ESBL Plasmids in Escherichia coli. Front Microbiol. 10: 2119.
Maeda, S., Ito, M., Ando, T., Ishimoto, Y., Fujisawa, Y., Takahashi, H., … Kato, S. (2006). Horizontal transfer of nonconjugative plasmids in a colony biofilm of Escherichia coli. FEMS Microbiol Lett. 255 (1): 115-120.
Marti, E., Variatza, E., Balcazar, J. L. (2014). The role of aquatic ecosystems as reservoirs of antibiotic resistance. Trends Microbiol. 22 (1): 36-41.
McClure, R., Balasubramanian, D., Sun, Y., Bobrovskyy, M., Sumby, P., Genco, C. A., … Tjaden, B. (2013). Computational analysis of bacterial RNA-Seq data. Nucleic Acids Res. 41 (14): e140-e140.
Mielich-Süss, B. & Lopez, D. (2015). Molecular mechanisms involved in Bacillus subtilis biofilm formation. Environ Microbiol. 17 (3): 555-565.
Ministerio de Salud. (2018). Plan nacional de respuesta a la resitencia a los antimicrobianos. Plan estratégico. Bogotá.
Ortiz-Mayorga, J. L., Pineda-Rodríguez, I. G., Dennis, R. J., Porras, A. (2019). Costos atribuidos a las infecciones asociadas con la atención en salud en un hospital de Colombia, 2011-2015. Biomédica. 39 (1): 102-112.
Palchevskiy, V. & Finkel, S. E. (2006). Escherichia coli competence gene homologs are essential for competitive fitness and the use of DNA as a nutrient. J Bacteriol. 188 (11): 3902-3910.
Pendleton, J. N., Gorman, S. P., Gilmore, B. F. (2013). Clinical relevance of the ESKAPE pathogens. Expert Rev Anti Infect Ther. 11 (3): 297-308.
Pfeltz, R. F., Schmidt, J. L., Wilkinson, B. J. (2001). A microdilution plating method for population analysis of antibiotic-resistant staphylococci. Microb Drug Resist. 7 (3): 289-295.
Posada-Perlaza, C. E., Ramírez-Rojas, A., Porras, P., Adu-Oppong, B., Botero-Coy, A.-M., Hernández, F., … Zambrano, M. M. (2019). Bogotá River anthropogenic contamination alters microbial communities and promotes spread of antibiotic resistance genes. Sci Rep. 9(1): 11764.
Pruesse, E., Quast, C., Knittel, K., Fuchs, B. M., Ludwig, W., Peplies, J., Glockner, F. O. (2007). SILVA: A comprehensive online resource for quality checked and aligned ribosomal RNA sequence data compatible with ARB. Nucleic Acids Res. 35 (21): 7188-7196.
Rada, A. M., Hernández-Gómez, C., Restrepo, E., Villegas, M. V. (2019). Distribución y caracterización molecular de betalactamasas en bacterias Gram negativas en Colombia, 2001-2016. Biomédica. 39: 199-220.
Rasheed, J. K., Anderson, G. J., Yigit, H., Queenan, A. M., Domenech-Sánchez, A., Swenson, J. M., … Tenover, F. C. (2000). Characterization of the Extended-Spectrum beta -Lactamase Reference Strain, Klebsiella pneumoniae K6 (ATCC 700603), Which Produces the Novel Enzyme SHV-18. Antimicrob Agents Chemother. 44 (9): 2382-2388.
Rizzo, L., Manaia, C., Merlin, C., Schwartz, T., Dagot, C., Ploy, M. C., … Fatta-Kassinos, D. (2013). Urban wastewater treatment plants as hotspots for antibiotic resistant bacteria and genes spread into the environment: A review. Sci Total Environ. 447: 345-360.
Rodríguez-Rojas, A., Rodríguez-Beltrán, J., Couce, A., Blázquez, J. (2013). Antibiotics and antibiotic resistance: A bitter fight against evolution. Int J Med Microbiol. 303 (6-7): 293-297.
Rojas, L. J., Mojica, M. F., Blanco, V. M., Correa, A., Montealegre, M. C., De La Cadena, E., … Villegas, M. V. (2013). Emergence of Klebsiella pneumoniae coharboring KPC and VIM carbapenemases in Colombia. Antimicrob Agents Chemother. 57: 1101-1102.
Shon, A. S., Bajwa, R. P. S., Russo, T. A. (2013). Hypervirulent (hypermucoviscous) Klebsiella pneumoniae. Virulence. 4 (2): 107-118.
Shun-Mei, E., Zeng, J.-M., Yuan, H., Lu, Y., Cai, R.-X., Chen, C. (2018). Sub-inhibitory concentrations of fluoroquinolones increase conjugation frequency. Microb Pathog. 114: 57-62.
Skippington, E. & Ragan, M. A. (2011). Lateral genetic transfer and the construction of genetic exchange communities. FEMS Microbiol Rev. 35 (5): 707-735.
Suescún, A. V., Cubillos, J. R., Zambrano, M. M. (2006). Genes involucrados en la biogénesis de fimbrias afectan la formación de biopelículas por parte de Klebsiella pneumoniae. Biomédica. 26: 528-537.
Sun, D., Zhang, Y., Mei, Y., Jiang, H., Xie, Z., Liu, H., … Shen, P. (2006). Escherichia coli is naturally transformable in a novel transformation system. FEMS Microbiol Lett. 265 (2): 249-255.
Van Laar, T. A., Chen, T., You, T., Leung, K. P. (2015). Sublethal Concentrations of Carbapenems Alter Cell Morphology and Genomic Expression of Klebsiella pneumoniae Biofilms. Antimicrob Agents Chemother. 59 (3): 1707-1717.
Venkatachalam, I., Teo, J., Balm, M. N. D., Fisher, D. A., Jureen, R., Lin, R. T. P. (2012). Klebsiella pneumoniae carbapenemase-producing enterobacteria in hospital, Singapore. Emerg Infect Dis. 18 (8): 1381-1383.
Ventola, C. L. (2015). The antibiotic resistance crisis: Part 1: Causes and threats. P T. 40 (4): 277-283. World Health Organization, WHO. (2016). WHO | Global action plan on AMR. Accessed on: July 2019. Retrieved from: https://www.who.int/antimicrobial-resistance/global-action-plan/en/Yanisch-Perron, C., Vieira, J., Messing, J. (1985). Improved M13 phage cloning vectors and host strains: Nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 33 (1): 103-119.
Zhang, Y., Shi, C., Yu, J., Ren, J., Sun, D. (2012). RpoS regulates a novel type of plasmid DNA transfer in Escherichia coli. PLoS One. 7 (3): e33514.
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