| Title | ddPCR applied on archived Continuous Plankton Recorder samples reveals long-term occurrence of class 1 integrons and a sulphonamide resistance gene in marine plankton communities |
| Abstract | Antibiotic resistance is a rising threat for human health. Although in clinical settings and terrestrial environments the rise of antibiotic resistant bacteria is well documented, their dissemination and spread in the marine environment, covering almost two-thirds of the Earth's surface, is still poorly understood. In this study, the presence and abundance of sulphonamide resistance gene (sul2) and class 1 integron-integrase gene (intI1), used as markers for the occurrence and spread of antibiotic resistance genes since the beginning of the antibiotic era, were investigated. Twenty-nine archived formalin-fixed samples, collected by the Continuous Plankton Recorder (CPR) survey in the Atlantic Ocean and North Sea from 1970 to 2011, were analysed using Droplet Digital PCR (ddPCR) applied for the first time on CPR samples. The two marker genes were present in a large fraction of the samples (48% for sul2 and 76% for intI1). In contrast, results from Real-Time PCR performed on the same samples greatly underestimate their occurrence (21% for sul2 and 52% for intI1). Overall, besides providing successful use of ddPCR for the molecular analysis of CPR samples, this study reveals long-term occurrence and spread of sul2 gene and class 1 integrons in the plankton-associated bacterial communities in the ocean. |
| Source | Environmental microbiology reports 10 (4), pp. 458–464 |
| Keywords | ddPCR |
| Journal | Environmental microbiology reports |
| Editor | Wiley-Blackwell, [Oxford], Stati Uniti d'America |
| Year | 2018 |
| Type | Articolo in rivista |
| DOI | 10.1111/1758-2229.12665 |
| Authors | Andrea Di Cesare, Sara Petrin, Diego Fontaneto, Carmen Losasso, Ester M. Eckert, Giovanni Tassistro, Alessio Borello, Antonia Ricci, William H. Wilson, Carla Pruzzo, Luigi Vezzulli |
| Text | 390469 2018 10.1111/1758 2229.12665 ddPCR ddPCR applied on archived Continuous Plankton Recorder samples reveals long term occurrence of class 1 integrons and a sulphonamide resistance gene in marine plankton communities Andrea Di Cesare, Sara Petrin, Diego Fontaneto, Carmen Losasso, Ester M. Eckert, Giovanni Tassistro, Alessio Borello, Antonia Ricci, William H. Wilson, Carla Pruzzo, Luigi Vezzulli 1Department of Earth, Environmental and Life Sciences DISTAV , University of Genoa, Genoa, Italy. 2O.U. Microbial Ecology, Department of Food Safety, Istituto Zooprofilattico Sperimentale delle Venezie, Legnaro, Italy. 3Microbial Ecology Group MEG , National Research Council Institute of Ecosystem Study CNR ISE , Verbania, Italy. 4CPR Survey, Marine Biological Association, The Laboratory, Citadel Hill, Plymouth, UK. Antibiotic resistance is a rising threat for human health. Although in clinical settings and terrestrial environments the rise of antibiotic resistant bacteria is well documented, their dissemination and spread in the marine environment, covering almost two thirds of the Earth s surface, is still poorly understood. In this study, the presence and abundance of sulphonamide resistance gene sul2 and class 1 integron integrase gene intI1 , used as markers for the occurrence and spread of antibiotic resistance genes since the beginning of the antibiotic era, were investigated. Twenty nine archived formalin fixed samples, collected by the Continuous Plankton Recorder CPR survey in the Atlantic Ocean and North Sea from 1970 to 2011, were analysed using Droplet Digital PCR ddPCR applied for the first time on CPR samples. The two marker genes were present in a large fraction of the samples 48% for sul2 and 76% for intI1 . In contrast, results from Real Time PCR performed on the same samples greatly underestimate their occurrence 21% for sul2 and 52% for intI1 . Overall, besides providing successful use of ddPCR for the molecular analysis of CPR samples, this study reveals long term occurrence and spread of sul2 gene and class 1 integrons in the plankton associated bacterial communities in the ocean. 10 Published version https //onlinelibrary.wiley.com/doi/abs/10.1111/1758 2229.12665 ddPCR 2018_EMIR_Di Cesare et al.pdf Articolo in rivista Wiley Blackwell 1758 2229 Environmental microbiology reports Environmental microbiology reports Environ. microbiol. rep. Environmental microbiology reports diego.fontaneto FONTANETO DIEGO andrea.dicesare DI CESARE ANDREA estermaria.eckert ECKERT ESTER MARIA |
