{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T00:11:16Z","timestamp":1790727076204,"version":"4.1.0"},"reference-count":40,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2021,9,4]],"date-time":"2021-09-04T00:00:00Z","timestamp":1630713600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100006537","name":"Campus France","doi-asserted-by":"publisher","award":["18G0819"],"award-info":[{"award-number":["18G0819"]}],"id":[{"id":"10.13039\/501100006537","id-type":"DOI","asserted-by":"publisher"}]},{"name":"IHU M\u00e9diterran\u00e9e Infection","award":["2020"],"award-info":[{"award-number":["2020"]}]},{"name":"Minist\u00e8re de l'enseignement sup\u00e9rieur et de la recherche scientifique Tunisie","award":["18G0819"],"award-info":[{"award-number":["18G0819"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>Hospital environments constitute the main reservoir of multidrug-resistant bacteria. In this study we aimed to investigate the presence of Gram-negative bacteria in one Northwestern Tunisian hospital environment, and characterize the genes involved in bacterial resistance. A total of 152 environmental isolates were collected from various surfaces and isolated using MacConkey medium supplemented with cefotaxime or imipenem, with 81 fermenter bacteria (27 Escherichia coli, and 54 Enterobacter spp., including 46 Enterobacter cloacae), and 71 non-fermenting bacteria (69 Pseudomonas spp., including 54 Pseudomonas aeruginosa, and 2 Stenotrophomonas maltophilia) being identified by the MALDI-TOF-MS method. Antibiotic susceptibility testing was performed by disk diffusion method and E-Test was used to determine MICs for imipenem. Several genes implicated in beta-lactams resistance were characterized by PCR and sequencing. Carbapenem resistance was detected among 12 isolates; nine E. coli (blaNDM-1 (n = 8); blaNDM-1 + blaVIM-2 (n = 1)) and three P. aeruginosa were carbapenem-resistant by loss of OprD porin. The whole-genome sequencing of P. aeruginosa 97H was determined using Illumina MiSeq sequencer, typed ST285, and harbored blaOXA-494. Other genes were also detected, notably blaTEM (n = 23), blaCTX-M-1 (n = 10) and blaCTX-M-9 (n = 6). These new epidemiological data imposed new surveillance strategies and strict hygiene rules to decrease the spread of multidrug-resistant bacteria in this area.<\/jats:p>","DOI":"10.3390\/diagnostics11091617","type":"journal-article","created":{"date-parts":[[2021,9,6]],"date-time":"2021-09-06T10:59:12Z","timestamp":1630925952000},"page":"1617","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Occurrence of NDM-1 and VIM-2 Co-Producing Escherichia coli and OprD Alteration in Pseudomonas aeruginosa Isolated from Hospital Environment Samples in Northwestern Tunisia"],"prefix":"10.3390","volume":"11","author":[{"given":"Raouaa","family":"Maaroufi","sequence":"first","affiliation":[{"name":"MEPHI, IHU M\u00e9diterran\u00e9e Infection, 19-21, Boulevard Jean Moulin, Aix-Marseille Universit\u00e9, 13005 Marseille, France"},{"name":"Laboratoire des Microorganismes et Biomol\u00e9cules Actives, Facult\u00e9 des Sciences de Tunis, Campus Universitaire, Universit\u00e9 de Tunis El-Manar, Tunis 2098, Tunisia"},{"name":"Laboratoire de Recherche Sciences et Technologies de l\u2019Environnement, Institut Sup\u00e9rieur des Sciences et Technologies de l\u2019Environnement de Borj-Cedria, Technop\u00f4le de Borj-Cedria, Universit\u00e9 de Carthage, PB 1003, Hammam-Lif 2050, Tunisia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Olfa","family":"Dziri","sequence":"additional","affiliation":[{"name":"Laboratoire des Microorganismes et Biomol\u00e9cules Actives, Facult\u00e9 des Sciences de Tunis, Campus Universitaire, Universit\u00e9 de Tunis El-Manar, Tunis 2098, Tunisia"},{"name":"Laboratoire de Recherche Sciences et Technologies de l\u2019Environnement, Institut Sup\u00e9rieur des Sciences et Technologies de l\u2019Environnement de Borj-Cedria, Technop\u00f4le de Borj-Cedria, Universit\u00e9 de Carthage, PB 1003, Hammam-Lif 2050, Tunisia"},{"name":"Unit\u00e9 de Service en Commun pour la Recherche \u00abPlateforme g\u00e9nomique\u00bb Institut Sup\u00e9rieur des Sciences et Technologies de l\u2019Environnement de Borj-Cedria, Technop\u00f4le de Borj-Cedria, Universit\u00e9 de Carthage, PB 1003, Hammam-Lif 2050, Tunisia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Linda","family":"Hadjadj","sequence":"additional","affiliation":[{"name":"MEPHI, IHU M\u00e9diterran\u00e9e Infection, 19-21, Boulevard Jean Moulin, Aix-Marseille Universit\u00e9, 13005 Marseille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6466-2324","authenticated-orcid":false,"given":"Seydina M.","family":"Diene","sequence":"additional","affiliation":[{"name":"MEPHI, IHU M\u00e9diterran\u00e9e Infection, 19-21, Boulevard Jean Moulin, Aix-Marseille Universit\u00e9, 13005 Marseille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jean-Marc","family":"Rolain","sequence":"additional","affiliation":[{"name":"MEPHI, IHU M\u00e9diterran\u00e9e Infection, 19-21, Boulevard Jean Moulin, Aix-Marseille Universit\u00e9, 13005 Marseille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8294-8289","authenticated-orcid":false,"given":"Chedly","family":"Chouchani","sequence":"additional","affiliation":[{"name":"Laboratoire des Microorganismes et Biomol\u00e9cules Actives, Facult\u00e9 des Sciences de Tunis, Campus Universitaire, Universit\u00e9 de Tunis El-Manar, Tunis 2098, Tunisia"},{"name":"Laboratoire de Recherche Sciences et Technologies de l\u2019Environnement, Institut Sup\u00e9rieur des Sciences et Technologies de l\u2019Environnement de Borj-Cedria, Technop\u00f4le de Borj-Cedria, Universit\u00e9 de Carthage, PB 1003, Hammam-Lif 2050, Tunisia"},{"name":"Unit\u00e9 de Service en Commun pour la Recherche \u00abPlateforme g\u00e9nomique\u00bb Institut Sup\u00e9rieur des Sciences et Technologies de l\u2019Environnement de Borj-Cedria, Technop\u00f4le de Borj-Cedria, Universit\u00e9 de Carthage, PB 1003, Hammam-Lif 2050, Tunisia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,9,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1016\/j.jhin.2009.03.030","article-title":"The role of hospital cleaning in the control of hospital-acquired infection","volume":"73","author":"Dancer","year":"2009","journal-title":"J. Hosp. Infect."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"59","DOI":"10.4103\/0300-1652.103543","article-title":"Healthcar\u2013associated infections: A public health problem","volume":"53","author":"Revelas","year":"2012","journal-title":"Niger. Med. J."},{"key":"ref_3","first-page":"1","article-title":"Epidemiology and risk factors for nosocomial infection in the respiratory intensive care unit of a teaching hospital in China: A prospective surveillance during 2013 and 2015","volume":"19","author":"Wang","year":"2019","journal-title":"BMC Infect. Dis."},{"key":"ref_4","first-page":"1039","article-title":"Update on Bacterial Nosocomial Infections","volume":"16","author":"Bereket","year":"2012","journal-title":"Eur. Rev. Med. Pharmacol. Sci."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1413","DOI":"10.1378\/chest.14-2171","article-title":"Antimicrobial resistance in hospital-acquired gram-negative bacterial infections","volume":"147","author":"Mehrad","year":"2015","journal-title":"Chest"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1309","DOI":"10.1056\/NEJMoa1914433","article-title":"Multidrug-Resistant Bacterial Infections in U.S. Hospitalized Patients, 2012\u20132017","volume":"382","author":"Jernigan","year":"2020","journal-title":"N. Engl. J. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1941","DOI":"10.3389\/fmicb.2019.01941","article-title":"Understanding the Epidemiology of Multi-Drug Resistant Gram-Negative Bacilli in the Middle East Using a One Health Approach","volume":"10","author":"Dandachi","year":"2019","journal-title":"Front. Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"S8","DOI":"10.1016\/S0924-8579(10)70004-2","article-title":"Emerging carbapenemases: A global perspective","volume":"36","author":"Walsh","year":"2010","journal-title":"Int. J. Antimicrob. Agents"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"725","DOI":"10.1111\/j.1600-0463.2011.02793.x","article-title":"VIM and IMP metallo-beta-lactamases and other extended-spectrum beta-lactamases in Escherichia coli and Klebsiella pneumoniae from environmental samples in a Tunisian hospital","volume":"119","author":"Chouchani","year":"2011","journal-title":"APMIS"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"399","DOI":"10.1089\/mdr.2015.0354","article-title":"Detection of CTX-M-15-Producing Escherichia coli Isolates of Lineages ST131-B2 and ST167-Ain Environmental Samples of a Tunisian Hospital","volume":"22","author":"Dziri","year":"2016","journal-title":"Microb. Drug Resist."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1016\/j.ijantimicag.2013.06.001","article-title":"ISPa46, a novel insertion sequence in the oprD porin gene of an imipenem-resistant Pseudomonas aeruginosa isolate from a cystic fibrosis patient in Marseille, France","volume":"42","author":"Diene","year":"2013","journal-title":"Int. J. Antimicrob. Agents"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1093\/jac\/dkl481","article-title":"Multiplex PCR for rapid detection of genes encoding acquired metallo-\u03b2-lactamases","volume":"59","author":"Ellington","year":"2007","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Roschanski, N., Fischer, J., Guerra, B., and Roesler, U. (2014). Development of a multiplex real-time PCR for the rapid detection of the predominant beta-lactamase genes CTX-M, SHV, TEM and CIT-type AmpCs in Enterobacteriaceae. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0100956"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"17","DOI":"10.2807\/1560-7917.ES.2018.23.6.17-00672","article-title":"Multiplex+ PCR for detection of plasmid-mediated colistin resistance determinants, mcr-1, mcr-2, mcr-3, mcr-4 and mcr-5 for surveillance purposes","volume":"23","author":"Rebelo","year":"2018","journal-title":"Euro. Surveill."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"167","DOI":"10.3109\/1040841X.2011.552880","article-title":"Evolution of beta-lactams resistance in Gram-negative bacteria in Tunisia","volume":"37","author":"Chouchani","year":"2011","journal-title":"Crit. Rev Microbiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"871","DOI":"10.1016\/j.ijantimicag.2018.06.002","article-title":"Metallo-\u03b2-lactamases and Class D Carbapenemases in South-East Tunisia: Implication of Mobile Genetic Elements in Their Dissemination","volume":"52","author":"Dziri","year":"2018","journal-title":"Int. J. Antimicrob. Agents"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1282","DOI":"10.1089\/mdr.2019.0115","article-title":"First Report of NDM and VIM Coproducing Klebsiella pneumoniae in Tunisia and Emergence of Novel Clones","volume":"25","author":"Dziri","year":"2019","journal-title":"Microb. Drug Resist."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"5046","DOI":"10.1128\/AAC.00774-09","article-title":"Characterization of a new metallo-beta-lactamase gene, bla (NDM-1), and a novel erythromycin esterase gene carried on a unique genetic structure in Klebsiella pneumoniae sequence type 14 from India","volume":"53","author":"Yong","year":"2009","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1699","DOI":"10.1111\/j.1469-0691.2010.03385.x","article-title":"New Delhi metallo-beta-lactamase (NDM-1): Towards a new pandemia?","volume":"12","author":"Rolain","year":"2010","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"20809","DOI":"10.2807\/1560-7917.ES2014.19.20.20809","article-title":"New Delhi Metallo-beta-lactamase around the world: An eReview using Google Maps","volume":"19","author":"Berrazeg","year":"2014","journal-title":"Euro. Surveill."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"3036","DOI":"10.1128\/AAC.00049-11","article-title":"Emergence of NDM-1-producing Enterobacteriaceae in Belgium","volume":"55","author":"Bogaerts","year":"2011","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"33","DOI":"10.2217\/fmb.13.69","article-title":"New Delhi metallo-\u03b2-lactamase-producing Acinetobacter baumannii: A novel paradigm for spreading antibiotic resistance genes","volume":"9","author":"Bonnin","year":"2014","journal-title":"Future Microbiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1087","DOI":"10.1128\/AAC.05620-11","article-title":"Tn125-Related Acquisition of blaNDM-Like Genes in Acinetobacter baumannii","volume":"56","author":"Poirel","year":"2012","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"371","DOI":"10.1016\/j.envpol.2017.05.095","article-title":"Abundance of carbapenemase genes (blaKPC, blaNDM and blaOXA-48) in wastewater effluents from Tunisian hospitals","volume":"229","author":"Nasri","year":"2017","journal-title":"Environ. Pollut."},{"key":"ref_25","first-page":"e25952","article-title":"Investigation of OprD Porin Protein Levels in Carbapenem-Resistant Pseudomonas aeruginosa Isolates","volume":"8","author":"Terzi","year":"2015","journal-title":"Jundishapur J. Microbiol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1772","DOI":"10.1093\/jac\/dkt098","article-title":"Complexity of resistance mechanisms to imipenem in intensive care unit strains of Pseudomonas aeruginosa","volume":"68","author":"Fournier","year":"2013","journal-title":"J. Antimicrob Chemother."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1763","DOI":"10.1093\/jac\/dkt115","article-title":"Diversity of b-lactam resistance mechanisms in cystic fibrosis isolates of Pseudomonas aeruginosa: A French multicentre study","volume":"68","author":"Llanes","year":"2013","journal-title":"J. Antimicrob Chemother."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"335","DOI":"10.1089\/mdr.2014.0235","article-title":"Emergence of Carbapenem-Resistant Pseudomonas aeruginosa and Acinetobacter baumannii Clinical Isolates Collected from Some Libyan Hospitals","volume":"21","author":"Mathlouthi","year":"2015","journal-title":"Microb. Drug Resist."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"529","DOI":"10.1016\/j.ijantimicag.2014.12.029","article-title":"Carbapenem resistance in cystic fibrosis strains of Pseudomonas aeruginosa as a result of amino acid substitutions in porin OprD","volume":"45","author":"Richardot","year":"2015","journal-title":"Int. J. Antimicrob. Agents"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2829","DOI":"10.1128\/JCM.00997-07","article-title":"Outbreak of Pseudomonas aeruginosa infections with PER-1 extended-spectrum \u03b2-lactamase in Warsaw, Poland: Further evidence for an international clonal complex","volume":"45","author":"Empel","year":"2007","journal-title":"J. Clin. Microbiol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"348","DOI":"10.1016\/S0399-077X(03)00180-X","article-title":"Bacteriological and susceptibility study of 1930 strains isolated from UTIs in a Tunis university hospital","volume":"33","author":"Larabi","year":"2003","journal-title":"Med. Malad. Infect."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"40208","DOI":"10.1038\/srep40208","article-title":"Mechanisms of intrinsic resistance and acquired susceptibility of Pseudomonas aeruginosa isolated from cystic fibrosis patients to temocillin, a revived antibiotic","volume":"7","author":"Chalhob","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"936","DOI":"10.1128\/AAC.01807-15","article-title":"Characterization of a Carbapenem hydrolyzing Enzyme, PoxB, in Pseudomonas aeruginosa PAO1","volume":"60","author":"Zincke","year":"2015","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2043","DOI":"10.1128\/AAC.48.6.2043-2048.2004","article-title":"Biochemical characterization of the naturally occurring oxacillinase OXA-50 of Pseudomonas aeruginosa","volume":"48","author":"Girlich","year":"2004","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1016\/j.gene.2005.05.027","article-title":"Characterization of poxB, a chromosomal encoded Pseudomonas aeruginosa oxacillinase","volume":"358","author":"Kong","year":"2005","journal-title":"Gene"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1016\/j.ijfoodmicro.2015.02.023","article-title":"Detection of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae in vegetables, soil and water of the farm environment in Tunisia","volume":"203","author":"Jouini","year":"2015","journal-title":"Int. J. Food Microbiol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1089\/mdr.2009.0900","article-title":"Characterization of a Novel Extended-Spectrum TEM-Type b-Lactamase, TEM-164, in a Clinical Strain of Klebsiella pneumoniae in Tunisia","volume":"15","author":"Mercuri","year":"2009","journal-title":"Microb. Drug. Resist."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"258","DOI":"10.4103\/0255-0857.83909","article-title":"First detection of TEM-116 extended-spectrum \u03b2-lactamase in a Providencia stuartii isolate from a Tunisian hospital","volume":"29","author":"Lahlaoui","year":"2011","journal-title":"Indian J. Med. Microbiol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"38","DOI":"10.3389\/fcimb.2015.00038","article-title":"Occurrence of blaCTX-M-1, qnrB1 and virulence genes in avian ESBL-producing Escherichia coli isolates from Tunisia","volume":"5","author":"Kilani","year":"2015","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1081","DOI":"10.1007\/s10096-011-1410-z","article-title":"Characterization of IncA\/C conjugative plasmids harboring blaTEM-52 and blaCTX-M-15 extended-spectrum \u03b2-lactamases in clinical isolates of Escherichia coli in Tunisia","volume":"31","author":"Chouchani","year":"2012","journal-title":"Eur. J. Clin. Microbiol. Infect. Dis."}],"container-title":["Diagnostics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-4418\/11\/9\/1617\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:56:28Z","timestamp":1760165788000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-4418\/11\/9\/1617"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,9,4]]},"references-count":40,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2021,9]]}},"alternative-id":["diagnostics11091617"],"URL":"https:\/\/doi.org\/10.3390\/diagnostics11091617","relation":{},"ISSN":["2075-4418"],"issn-type":[{"value":"2075-4418","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,9,4]]}}}