{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,16]],"date-time":"2026-06-16T06:30:56Z","timestamp":1781591456730,"version":"3.54.5"},"reference-count":50,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2021,5,10]],"date-time":"2021-05-10T00:00:00Z","timestamp":1620604800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2021,5,10]],"date-time":"2021-05-10T00:00:00Z","timestamp":1620604800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Commun Biol"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The increasing prevalence of antimicrobial resistance has become a global health problem. <jats:italic>Acinetobacter baumannii<\/jats:italic> is an important nosocomial pathogen due to its capacity to persist in the hospital environment. It has a high mortality rate and few treatment options. Antibiotic combinations can help to fight multi-drug resistant (MDR) bacterial infections, but they are rarely used in the clinics and mostly unexplored. The interaction between bacteriostatic and bactericidal antibiotics are mostly reported as antagonism based on the results obtained in the susceptible model laboratory strain <jats:italic>Escherichia coli<\/jats:italic>. However, in the present study, we report a synergistic interaction between nalidixic acid and tetracycline against clinical multi-drug resistant <jats:italic>A. baumannii<\/jats:italic> and <jats:italic>E. coli<\/jats:italic>. Here we provide mechanistic insight into this dichotomy. The synergistic combination was studied by checkerboard assay and time-kill curve analysis. We also elucidate the mechanism behind this synergy using several techniques such as fluorescence spectroscopy, flow cytometry, fluorescence microscopy, morphometric analysis, and real-time polymerase chain reaction. Nalidixic acid and tetracycline combination displayed synergy against most of the MDR clinical isolates of <jats:italic>A. baumannii<\/jats:italic> and <jats:italic>E. coli<\/jats:italic> but not against susceptible isolates. Finally, we demonstrate that this combination is also effective in vivo in an <jats:italic>A. baumannii<\/jats:italic>\/<jats:italic>Caenorhabditis elegans<\/jats:italic> infection model (<jats:italic>p<\/jats:italic>\u2009&lt;\u20090.001)<\/jats:p>","DOI":"10.1038\/s42003-021-02074-5","type":"journal-article","created":{"date-parts":[[2021,5,10]],"date-time":"2021-05-10T10:04:16Z","timestamp":1620641056000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":32,"title":["Mechanistic insights into synergy between nalidixic acid and tetracycline against clinical isolates of Acinetobacter baumannii and Escherichia coli"],"prefix":"10.1038","volume":"4","author":[{"given":"Amit","family":"Gaurav","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Varsha","family":"Gupta","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sandeep K.","family":"Shrivastava","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7965-8176","authenticated-orcid":false,"given":"Ranjana","family":"Pathania","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,5,10]]},"reference":[{"key":"2074_CR1","unstructured":"WHO. Global Priority List of Antibiotic-Resistant Bacteria to Guide Research, Discovery, and Development of New Antibiotics (WHO, 2017)."},{"key":"2074_CR2","doi-asserted-by":"publisher","first-page":"409","DOI":"10.1128\/CMR.00058-16","volume":"30","author":"D Wong","year":"2017","unstructured":"Wong, D. et al. Clinical and pathophysiological overview of Acinetobacter infections: a century of challenges. Clin. Microbiol. Rev. 30, 409\u2013447 (2017).","journal-title":"Clin. Microbiol. Rev."},{"key":"2074_CR3","unstructured":"Bush, K. in Topics in Medicinal Chemistry - Antibacterials: Volume I (eds Fisher, J. F., Mobashery, S. & Miller, M. J.) 69\u201388 (Springer, 2018)."},{"key":"2074_CR4","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.mib.2015.05.008","volume":"27","author":"T Bollenbach","year":"2015","unstructured":"Bollenbach, T. Antimicrobial interactions: mechanisms and implications for drug discovery and resistance evolution. Curr. Opin. Microbiol. 27, 1\u20139 (2015).","journal-title":"Curr. Opin. Microbiol."},{"key":"2074_CR5","doi-asserted-by":"publisher","first-page":"668","DOI":"10.1038\/nature05685","volume":"446","author":"R Chait","year":"2007","unstructured":"Chait, R., Craney, A. & Kishony, R. Antibiotic interactions that select against resistance. Nature 446, 668 (2007).","journal-title":"Nature"},{"key":"2074_CR6","doi-asserted-by":"publisher","first-page":"31","DOI":"10.1016\/j.mib.2020.05.012","volume":"57","author":"GJ Sullivan","year":"2020","unstructured":"Sullivan, G. J., Delgado, N. N., Maharjan, R. & Cain, A. K. How antibiotics work together: molecular mechanisms behind combination therapy. Curr. Opin. Microbiol. 57, 31\u201340 (2020).","journal-title":"Curr. Opin. Microbiol."},{"key":"2074_CR7","doi-asserted-by":"publisher","first-page":"301","DOI":"10.1001\/archinte.1952.00240090022003","volume":"90","author":"E Jawetz","year":"1952","unstructured":"Jawetz, E. Antibiotic synergism and antagonism: review of experimental evidence. AMA Arch. Intern. Med. 90, 301\u2013309 (1952).","journal-title":"AMA Arch. Intern. Med."},{"key":"2074_CR8","doi-asserted-by":"publisher","first-page":"348","DOI":"10.1038\/nchembio.559","volume":"7","author":"L Ejim","year":"2011","unstructured":"Ejim, L. et al. Combinations of antibiotics and nonantibiotic drugs enhance antimicrobial efficacy. Nat. Chem. Biol. 7, 348 (2011).","journal-title":"Nat. Chem. Biol."},{"key":"2074_CR9","doi-asserted-by":"publisher","first-page":"139","DOI":"10.1128\/CMR.1.2.139","volume":"1","author":"GM Eliopoulos","year":"1988","unstructured":"Eliopoulos, G. M. & Eliopoulos, C. T. Antibiotic combinations: should they be tested? Clin. Microbiol. Rev. 1, 139 (1988).","journal-title":"Clin. Microbiol. Rev."},{"key":"2074_CR10","doi-asserted-by":"publisher","first-page":"259","DOI":"10.1038\/s41586-018-0278-9","volume":"559","author":"AR Brochado","year":"2018","unstructured":"Brochado, A. R. et al. Species-specific activity of antibacterial drug combinations. Nature 559, 259\u2013263 (2018).","journal-title":"Nature"},{"key":"2074_CR11","doi-asserted-by":"publisher","first-page":"872","DOI":"10.15252\/msb.20156777","volume":"12","author":"S Chandrasekaran","year":"2016","unstructured":"Chandrasekaran, S. et al. Chemogenomics and orthology-based design of antibiotic combination therapies. Mol. Syst. Biol. 12, 872\u2013872 (2016).","journal-title":"Mol. Syst. Biol."},{"key":"2074_CR12","doi-asserted-by":"publisher","first-page":"4573","DOI":"10.1128\/AAC.02463-14","volume":"58","author":"PS Ocampo","year":"2014","unstructured":"Ocampo, P. S. et al. Antagonism between bacteriostatic and bactericidal antibiotics is prevalent. Antimicrob. Agents Chemother. 58, 4573 (2014).","journal-title":"Antimicrob. Agents Chemother."},{"key":"2074_CR13","doi-asserted-by":"publisher","first-page":"193","DOI":"10.1016\/j.ebiom.2019.07.041","volume":"46","author":"N Dillon","year":"2019","unstructured":"Dillon, N. et al. Surprising synergy of dual translation inhibition vs. Acinetobacter baumannii and other multidrug-resistant bacterial pathogens. Ebiomedicine 46, 193\u2013201 (2019).","journal-title":"Ebiomedicine"},{"key":"2074_CR14","doi-asserted-by":"publisher","first-page":"4678","DOI":"10.1128\/AAC.00497-10","volume":"54","author":"MM Sopirala","year":"2010","unstructured":"Sopirala, M. M. et al. Synergy testing by Etest, microdilution checkerboard, and time-kill methods for pan-drug-resistant Acinetobacter baumannii. Antimicrob. Agents Chemother. 54, 4678 (2010).","journal-title":"Antimicrob. Agents Chemother."},{"key":"2074_CR15","doi-asserted-by":"publisher","first-page":"72","DOI":"10.1016\/j.mib.2019.10.008","volume":"51","author":"MI Hutchings","year":"2019","unstructured":"Hutchings, M. I., Truman, A. W. & Wilkinson, B. Antibiotics: past, present and future. Curr. Opin. Microbiol. 51, 72\u201380 (2019).","journal-title":"Curr. Opin. Microbiol."},{"key":"2074_CR16","doi-asserted-by":"publisher","unstructured":"Shang, D. et al. Synergistic antibacterial activity of designed Trp-containing antibacterial peptides in combination with antibiotics against multidrug-resistant Staphylococcus epidermidis. Front. Microbiol. 10, https:\/\/doi.org\/10.3389\/fmicb.2019.02719 (2019).","DOI":"10.3389\/fmicb.2019.02719"},{"key":"2074_CR17","doi-asserted-by":"publisher","first-page":"1168","DOI":"10.1016\/j.chembiol.2013.07.006","volume":"20","author":"MA Farha","year":"2013","unstructured":"Farha, M. A., Verschoor, C. P., Bowdish, D. & Brown, E. D. Collapsing the proton motive force to identify synergistic combinations against Staphylococcus aureus. Chem. Biol. 20, 1168\u20131178 (2013).","journal-title":"Chem. Biol."},{"key":"2074_CR18","doi-asserted-by":"publisher","first-page":"e02792","DOI":"10.1128\/AEM.02792-18","volume":"85","author":"JM Griffith","year":"2019","unstructured":"Griffith, J. M. et al. Experimental evolution of Escherichia coli K-12 in the presence of proton motive force (PMF) uncoupler carbonyl cyanide-chlorophenylhydrazone selects for mutations affecting PMF-driven drug efflux pumps. Appl. Environ. Microbiol. 85, e02792\u201302718 (2019).","journal-title":"Appl. Environ. Microbiol."},{"key":"2074_CR19","doi-asserted-by":"publisher","first-page":"797","DOI":"10.1016\/j.cell.2007.06.049","volume":"130","author":"MA Kohanski","year":"2007","unstructured":"Kohanski, M. A., Dwyer, D. J., Hayete, B., Lawrence, C. A. & Collins, J. J. A common mechanism of cellular death induced by bactericidal antibiotics. Cell 130, 797\u2013810 (2007).","journal-title":"Cell"},{"key":"2074_CR20","doi-asserted-by":"crossref","unstructured":"Dwyer, D. J., Kohanski, M. A., Hayete, B. & Collins, J. J. Gyrase inhibitors induce an oxidative damage cellular death pathway in Escherichia coli. Mol. Syst. Biol. 3, 91 (2007).","DOI":"10.1038\/msb4100135"},{"key":"2074_CR21","doi-asserted-by":"publisher","first-page":"16169","DOI":"10.1073\/pnas.1311066110","volume":"110","author":"P Nonejuie","year":"2013","unstructured":"Nonejuie, P., Burkart, M., Pogliano, K. & Pogliano, J. Bacterial cytological profiling rapidly identifies the cellular pathways targeted by antibacterial molecules. Proc. Natl Acad. Sci. USA 110, 16169 (2013).","journal-title":"Proc. Natl Acad. Sci. USA"},{"key":"2074_CR22","doi-asserted-by":"publisher","first-page":"707","DOI":"10.1016\/j.cell.2009.10.025","volume":"139","author":"T Bollenbach","year":"2009","unstructured":"Bollenbach, T., Quan, S., Chait, R. & Kishony, R. Nonoptimal microbial response to antibiotics underlies suppressive drug interactions. Cell 139, 707\u2013718 (2009).","journal-title":"Cell"},{"key":"2074_CR23","doi-asserted-by":"publisher","first-page":"e0144697","DOI":"10.1371\/journal.pone.0144697","volume":"10","author":"M Maeda","year":"2016","unstructured":"Maeda, M., Shimada, T. & Ishihama, A. Strength and regulation of seven rRNA promoters in Escherichia coli. PLos ONE 10, e0144697 (2016).","journal-title":"PLos ONE"},{"key":"2074_CR24","doi-asserted-by":"publisher","first-page":"1728","DOI":"10.1128\/AAC.04198-14","volume":"59","author":"E Jayamani","year":"2015","unstructured":"Jayamani, E. et al. Insect-derived cecropins display activity against Acinetobacter baumannii in a whole-animal high-throughput Caenorhabditis elegans model. Antimicrob. Agents Chemother. 59, 1728\u20131737 (2015).","journal-title":"Antimicrob. Agents Chemother."},{"key":"2074_CR25","doi-asserted-by":"publisher","first-page":"162","DOI":"10.1038\/s42003-020-0877-5","volume":"3","author":"Y Liu","year":"2020","unstructured":"Liu, Y. et al. Anti-HIV agent azidothymidine decreases Tet(X)-mediated bacterial resistance to tigecycline in Escherichia coli. Commun. Biol. 3, 162 (2020).","journal-title":"Commun. Biol."},{"key":"2074_CR26","doi-asserted-by":"publisher","first-page":"11131","DOI":"10.1038\/s41598-018-29313-w","volume":"8","author":"L Zou","year":"2018","unstructured":"Zou, L. et al. Synergistic antibacterial activity of silver with antibiotics correlating with the upregulation of the ROS production. Sci. Rep. 8, 11131 (2018).","journal-title":"Sci. Rep."},{"key":"2074_CR27","doi-asserted-by":"publisher","first-page":"700","DOI":"10.1038\/msb.2013.57","volume":"9","author":"V L\u00e1z\u00e1r","year":"2013","unstructured":"L\u00e1z\u00e1r, V. et al. Bacterial evolution of antibiotic hypersensitivity. Mol. Syst. Biol. 9, 700 (2013).","journal-title":"Mol. Syst. Biol."},{"key":"2074_CR28","doi-asserted-by":"crossref","unstructured":"Cokol, M. et al. Systematic exploration of synergistic drug pairs. Mol. Syst. Biol. 7, 544 (2011).","DOI":"10.1038\/msb.2011.71"},{"key":"2074_CR29","doi-asserted-by":"publisher","first-page":"1073","DOI":"10.1016\/j.celrep.2014.02.007","volume":"6","author":"KB Wood","year":"2014","unstructured":"Wood, K. B., Wood, K. C., Nishida, S. & Cluzel, P. Uncovering scaling laws to infer multidrug response of resistant microbes and cancer cells. Cell Rep. 6, 1073\u20131084 (2014).","journal-title":"Cell Rep."},{"key":"2074_CR30","doi-asserted-by":"publisher","first-page":"547","DOI":"10.1016\/j.tibtech.2007.09.004","volume":"25","author":"G Cottarel","year":"2007","unstructured":"Cottarel, G. & Wierzbowski, J. Combination drugs, an emerging option for antibacterial therapy. Trends Biotechnol. 25, 547\u2013555 (2007).","journal-title":"Trends Biotechnol."},{"key":"2074_CR31","doi-asserted-by":"publisher","first-page":"193","DOI":"10.1007\/BF01956870","volume":"10","author":"TB Vree","year":"1988","unstructured":"Vree, T. B., Wijnands, W. J., Baars, A. M. & Hekster, Y. A. Pharmacokinetics of nalidixic acid in man: hydroxylation and glucuronidation. Pharm. Weekbl. Sci. 10, 193\u2013199 (1988).","journal-title":"Pharm. Weekbl. Sci."},{"key":"2074_CR32","unstructured":"USFDA. NegGram.pdf. NDA 14-214\/S-058, 3\u201312 (USFDA, 2008)."},{"key":"2074_CR33","doi-asserted-by":"publisher","first-page":"S40","DOI":"10.1093\/infdis\/131.Supplement.S40","volume":"131","author":"DM Musher","year":"1975","unstructured":"Musher, D. M., Minuth, J. N., Thorsteinsson, S. B. & Holmes, T. Effectiveness of achievable urinary concentrations of tetracyclines against \u201ctetracycline-resistant\u201d pathogenic bacteria. J. Infect. Dis. 131, S40\u2013S44 (1975).","journal-title":"J. Infect. Dis."},{"key":"2074_CR34","doi-asserted-by":"publisher","first-page":"106022","DOI":"10.1016\/j.ijantimicag.2020.106022","volume":"56","author":"A Gaurav","year":"2020","unstructured":"Gaurav, A., Kothari, A., Omar, B. J. & Pathania, R. Assessment of polymyxin B-doxycycline in combination against Pseudomonas aeruginosa in vitro and in a mouse model of acute pneumonia. Int. J. Antimicrob. Agents 56, 106022 (2020).","journal-title":"Int. J. Antimicrob. Agents"},{"key":"2074_CR35","doi-asserted-by":"publisher","first-page":"12","DOI":"10.1039\/C7MD00528H","volume":"9","author":"RJ Melander","year":"2018","unstructured":"Melander, R. J., Zurawski, D. V. & Melander, C. Narrow-spectrum antibacterial agents. Medchemcomm 9, 12\u201321 (2018).","journal-title":"Medchemcomm"},{"key":"2074_CR36","doi-asserted-by":"publisher","first-page":"156","DOI":"10.4065\/mcp.2010.0639","volume":"86","author":"S Leekha","year":"2011","unstructured":"Leekha, S., Terrell, C. L. & Edson, R. S. General principles of antimicrobial therapy. Mayo Clin. Proc. 86, 156\u2013167 (2011).","journal-title":"Mayo Clin. Proc."},{"key":"2074_CR37","doi-asserted-by":"publisher","first-page":"623","DOI":"10.1038\/nature25979","volume":"555","author":"L Maier","year":"2018","unstructured":"Maier, L. et al. Extensive impact of non-antibiotic drugs on human gut bacteria. Nature 555, 623\u2013628 (2018).","journal-title":"Nature"},{"key":"2074_CR38","unstructured":"CLSI. Performance Standards for Antimicrobial Susceptibility Testing (Clinical and Laboratory Standards Institute, 2018)."},{"key":"2074_CR39","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1093\/jac\/dkg301","volume":"52","author":"FC Odds","year":"2003","unstructured":"Odds, F. C. Synergy, antagonism, and what the chequerboard puts between them. J. Antimicrobial Chemother. 52, 1\u20131 (2003).","journal-title":"J. Antimicrobial Chemother."},{"key":"2074_CR40","doi-asserted-by":"publisher","first-page":"4353","DOI":"10.1038\/s41598-017-04462-6","volume":"7","author":"LP Menzel","year":"2017","unstructured":"Menzel, L. P. et al. Potent in vitro and in vivo antifungal activity of a small molecule host defense peptide mimic through a membrane-active mechanism. Sci. Rep. 7, 4353 (2017).","journal-title":"Sci. Rep."},{"key":"2074_CR41","doi-asserted-by":"publisher","first-page":"330","DOI":"10.1038\/nrmicro2549","volume":"9","author":"TA Krulwich","year":"2011","unstructured":"Krulwich, T. A., Sachs, G. & Padan, E. Molecular aspects of bacterial pH sensing and homeostasis. Nat. Rev. Microbiol. 9, 330\u2013343 (2011).","journal-title":"Nat. Rev. Microbiol."},{"key":"2074_CR42","doi-asserted-by":"publisher","first-page":"29","DOI":"10.3389\/fcell.2016.00029","volume":"4","author":"JD te Winkel","year":"2016","unstructured":"te Winkel, J. D., Gray, D. A., Seistrup, K. H., Hamoen, L. W. & Strahl, H. Analysis of antimicrobial-triggered membrane depolarization using voltage sensitive dyes. Front. Cell Dev. Biol. 4, 29 (2016).","journal-title":"Front. Cell Dev. Biol."},{"key":"2074_CR43","doi-asserted-by":"publisher","first-page":"57","DOI":"10.1007\/978-1-60761-411-1_4","volume":"594","author":"E Eruslanov","year":"2010","unstructured":"Eruslanov, E. & Kusmartsev, S. Identification of ROS using oxidized DCFDA and flow-cytometry. Methods Mol. Biol. 594, 57\u201372 (2010).","journal-title":"Methods Mol. Biol."},{"key":"2074_CR44","doi-asserted-by":"publisher","first-page":"e45","DOI":"10.1093\/nar\/29.9.e45","volume":"29","author":"MW Pfaffl","year":"2001","unstructured":"Pfaffl, M. W. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 29, e45 (2001).","journal-title":"Nucleic Acids Res."},{"key":"2074_CR45","unstructured":"Pfaffl, M. W. & Bustin, S. in AZ of Quantitative PCR 87\u2013120 (International Univ. Line, 2004)."},{"key":"2074_CR46","doi-asserted-by":"publisher","first-page":"685","DOI":"10.1007\/s10482-015-0524-1","volume":"108","author":"DJP Rocha","year":"2015","unstructured":"Rocha, D. J. P., Santos, C. S. & Pacheco, L. G. C. Bacterial reference genes for gene expression studies by RT-qPCR: survey and analysis. Antonie Van Leeuwenhoek 108, 685\u2013693 (2015).","journal-title":"Antonie Van Leeuwenhoek"},{"key":"2074_CR47","doi-asserted-by":"publisher","first-page":"e00968","DOI":"10.1128\/mBio.00968-17","volume":"8","author":"X Wang-Kan","year":"2017","unstructured":"Wang-Kan, X. et al. Lack of AcrB efflux function confers loss of virulence on Salmonella enterica Serovar Typhimurium. Mbio 8, e00968\u201300917 (2017).","journal-title":"Mbio"},{"key":"2074_CR48","doi-asserted-by":"publisher","first-page":"5421","DOI":"10.1046\/j.1432-1327.2000.01606.x","volume":"267","author":"J O\u2019brien","year":"2000","unstructured":"O\u2019brien, J., Wilson, I., Orton, T. & Pognan, F. Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity. Eur. J. Biochem. 267, 5421\u20135426 (2000).","journal-title":"Eur. J. Biochem."},{"key":"2074_CR49","doi-asserted-by":"crossref","unstructured":"Liu, Y. et al. Metformin restores tetracyclines susceptibility against multidrug resistant bacteria. Adv. Sci. 7, 1902227 (2020).","DOI":"10.1002\/advs.201902227"},{"key":"2074_CR50","doi-asserted-by":"publisher","first-page":"e01076","DOI":"10.1128\/mBio.01076-14","volume":"5","author":"AC Jacobs","year":"2014","unstructured":"Jacobs, A. C. et al. AB5075, a highly virulent isolate of Acinetobacter baumannii as a model strain for the evaluation of pathogenesis and antimicrobial treatments. Mbio 5, e01076\u201301014 (2014).","journal-title":"Mbio"}],"container-title":["Communications Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s42003-021-02074-5.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s42003-021-02074-5","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s42003-021-02074-5.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,2,1]],"date-time":"2023-02-01T08:15:47Z","timestamp":1675239347000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s42003-021-02074-5"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,5,10]]},"references-count":50,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2021,12]]}},"alternative-id":["2074"],"URL":"https:\/\/doi.org\/10.1038\/s42003-021-02074-5","relation":{},"ISSN":["2399-3642"],"issn-type":[{"value":"2399-3642","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,5,10]]},"assertion":[{"value":"1 July 2020","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"1 April 2021","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"10 May 2021","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"542"}}