Validation of quantitative real-time polymerase chain reaction for detection of Legionella pneumophila in hospital water networks.

BACKGROUND: Rapid monitoring of Legionella pneumophila (Lp) is essential to reduce the risk of Legionnaires' disease in healthcare facilities. However, culture results take at least eight days, delaying the implementation of corrective measures. Here, we assessed the performance of a qPCR method and determined qPCR action thresholds for the detection of Lp in hospital hot water networks (HWNs). METHODS: Hot water samples (N = 459) were collected from a hospital HWN. Lp were quantified using iQ-Check® Quanti real-time PCR Quantification kits (Bio-Rad) and the results were compared with those of culture. qPCR thresholds corresponding to the culture action thresholds of 10 and 1000 cfu/L were determined on a training dataset and validated on an independent dataset. RESULTS: Lp concentrations measured by culture and qPCR were correlated for both the training dataset (Spearman's correlation coefficient ρ = 0.687, P<0.0001) and the validation dataset (ρ = 0.661, P<0.0001). Lp qPCR positivity thresholds corresponding to culture action thresholds of 10 cfu/L was 91 genome units (gu) per litre (sensitivity, 86.4%; negative predictive value - NPV, 93.3%) and that corresponding to culture action thresholds of 1000 cfu/L was 1048 gu/L (sensitivity, 100%; NPV, 100%). CONCLUSION: Detection of Lp by qPCR could be implemented with confidence in hospitals as a complement to culture in the monitoring strategy to speed up the implementation of corrective measures.

Genome-based typing reveals rare events of patient contamination with Pseudomonas aeruginosa from other patients and sink traps in a medical intensive care unit.

AIM: We used genome-based typing data with the aim of identifying the routes of acquisition of Pseudomonas aeruginosa by patients hospitalized in a medical intensive care unit (MICU) over a long period in a non-epidemic context. METHODS: This monocentric prospective study took place over 10 months in 2019 in a 15-bed MICU that applies standard precautions of hygiene. Lockable sink traps installed at all water points of use were bleach disinfected twice a week. We sampled all sink traps weekly to collect 404 P. aeruginosa environmental isolates and collected all P. aeruginosa isolates (N = 115) colonizing or infecting patients (N = 65). All isolates had their phenotypic resistance profile determined and their genome sequenced, from which we identified resistance determinants and assessed the population structure of the collection at the nucleotide level to identify events of P. aeruginosa transmission. FINDINGS: All sink traps were positive for P. aeruginosa, each sink trap being colonized for several months by one or more clones. The combination of genomic and spatiotemporal data identified one potential event of P. aeruginosa transmission from a sink trap to a patient (1/65, 1.5%) and six events of patient cross-transmission, leading to the contamination of five patients (5/65, 7.7%). All transmitted isolates were fully susceptible to ß-lactams and aminoglycosides. CONCLUSIONS: Genome-based typing revealed the contamination of patients by P. aeruginosa originating from sink traps to be infrequent (1.5%) in an MICU with sink trap-bleaching measures, and that only 7.7% of the patients acquired P. aeruginosa originating from another patient.

Antibiotic resistance in elderly patients: Comparison of Enterobacterales causing urinary tract infections between community, nursing homes and hospital settings.

OBJECTIVE: The objective was to compare the prevalence of antibiotic resistance of, Escherichia coli, Proteus mirabilis, and Klebsiella pneumoniae in elderly patients in, three sectors: community, nursing homes, and hospital settings. MATERIAL AND METHODS: This study was a retrospective observational study conducted in, Bourgogne Franche-Comté (France). We collected positive urine samples from, patients over 75 years of age from six private laboratories. RESULTS: Antibiotic resistance rate for E. coli in nursing homes was close to that of the, ommunity setting. Conversely, resistance of K. pneumoniae in nursing homes was, close to hospital settings. No difference in resistance of P. mirabilis was observed, between the three healthcare sectors. CONCLUSIONS: Patients living in nursing homes should not be considered more at risk of, infection by multi-drug resistant E. coli than patients living in community setting. Screening of multi-drug resistant K. pneumoniae could be of interest for nursing home, patients.

[What academic training for pharmacy students for clinical pharmacy? Example of shared medication report]. / Quelle formation académique pour les nouvelles missions de pharmacie clinique à l'officine ? Exemple du bilan partagé de médication du sujet âgé.

OBJECTIVES: In 2018, the implementation of shared medication reports in pharmacy encourages pharmacists to cooperate with other healthcare professionals. This job allows a decrease of medication errors in elderly. This requires a reorganization of the training offered by universities (initial and continuing training). The aim is to present the results of this pedagogical experimentation. METHODS: The experimentation (years 2017-2018 and 2018-2019) required the creation of a course to allow students to carry out a pharmaceutical analysis suitable to elderly people, to set up and carry out a shared medication report in pharmacy. Then, during their 6th year internship, students had to carry out at least one shared medication report per month. A monthly follow-up was organized with a database online. RESULTS: Sixty-four students and 35 internship supervisors participated in the experimentation. All the students improved their ease in using clinical pharmacy tools (pharmaceutical analysis, pharmaceutical interventions, assessment of adherence, etc.). They carried out 345 shared medication reports. In 24.3% of cases, an improvement in the prescription was proposed to the doctor (general practitioner or specialist). For 80% of the internship supervisors, the initial training of the students helped to set up this new pharmacy activity. CONCLUSIONS: This teaching is appreciated by students and internship supervisors. It enabled the adoption of the various tools essential for carrying out shared medication reports in pharmacy. Shared medication reports reinforce the multidisciplinary work of pharmacists, especially with general practitioners.

Influenza vaccination coverage at community pharmacies: Positioning, needs, expectations and involvement of community pharmacists in the Franche-Comté region.

INTRODUCTION: Influenza vaccination coverage currently remains below the 75% recommended threshold by the World Health Organization. To correct this situation, experiments have been successively carried out in France to enable community pharmacists to vaccinate at-risk populations. In this context, a study was conducted with pharmacists from the French Franche-Comté region to evaluate their positioning, needs and expectations regarding influenza vaccination at community pharmacies. MATERIALS AND METHODS: A survey was created and sent to licensed pharmacists in March of 2018. This consisted of 4 parts: characteristics of the community pharmacy; positioning of the pharmacist regarding vaccinations carried out at the pharmacy; training needs and expectations; and willingness to implement vaccinations. RESULTS: The participation rate in this survey was 32% (137/427). More than 90% of the pharmacists agreed that community pharmacies' assets were adequate for the implementation of these vaccinations (accessibility and availability), although 52% considered this complicated. Their main fears were reluctance from patients and conflicts of interest with other health professionals authorized to vaccinate (58%). The needs and expectations regarding pharmacy student training were essential for 94% of them as well as continuous training of practicing pharmacists (96%). The willingness of pharmacists to vaccinate stemmed from the fact that influenza vaccination coverage would increase for at-risk subjects (36%). CONCLUSION: This survey allowed us to assess the favorable positioning and the real interest of pharmacists from Franche-Comté regarding the influenza vaccination done at community pharmacies, given the proviso that they were given relevant training and allocated adequate resources.

[Implementation of prehospital transfusion in case of trauma shock]. / Instauration d'un protocole de transfusion préhospitalière dans le choc hémorragique traumatique.

Transfusion plays a major role in the management of hemorrhagic shock where every minute counts. A pre-hospital transfusion protocol is established in the medical vehicle within the emergency department of CHR hospital Liège. It is based on predefined clinical and biological severity criteria which allow us to start a massive transfusion protocol as early as possible and thus to optimize its effect during traumatic hemorrhagic shock.

La transfusion occupe une place majeure dans la prise en charge du choc hémorragique où chaque minute compte. Un protocole de transfusion préhospitalière est instauré via le véhicule SMUR (Service Mobile d'Urgence-Réanimation) au sein du service des urgences du CHR de Liège. Il est basé sur des critères de gravité cliniques et biologiques prédéfinis qui permettent de débuter, le plus précocement possible, un protocole de transfusion massive et, ainsi, d'optimaliser son effet lors de choc hémorragique traumatique.

Comparison of pulsed-field gel electrophoresis and whole-genome-sequencing-based typing confirms the accuracy of pulsed-field gel electrophoresis for the investigation of local Pseudomonas aeruginosa outbreaks.

AIM: To determine whether pulsed-field gel electrophoresis (PFGE) accurately recognizes isolates belonging to clusters defined by techniques based on whole-genome sequencing (WGS) using Pseudomonas aeruginosa as a model. METHODS: We selected 65 isolates of ST395 P. aeruginosa isolated in seven European hospitals between 1998 and 2012. Isolates were typed by PFGE and sequenced by WGS. A core genome multi-locus sequence typing (cgMLST) analysis based on 3831 genes was performed with a homemade pipeline. FINDINGS: PFGE identified eight pulsotypes and cgMLST differentiated nine clusters and nine singletons. Five cgMLST clusters and pulsotypes (31/65 isolates) coincided perfectly. Isolates without evident epidemiological links grouped by PFGE were separated by cgMLST (16/65 isolates) differentiating cities, suggesting that PFGE should be kept for the investigation of local outbreaks. Importantly, hypermutator isolates still shared the pulsotype with their parents (16/65 isolates), whereas they were not recognized by cgMLST. This shows that PFGE was less affected than WGS-based typing by the accelerated genetic drift that occurs in epidemic P. aeruginosa. CONCLUSIONS: although WGS-based typing has logically become the new reference standard, we show here that the PFGE can be used with confidence for the investigation of local outbreaks caused by P. aeruginosa.

Exogenous acquisition of Pseudomonas aeruginosa in intensive care units: a prospective multi-centre study (DYNAPYO study).

BACKGROUND: Pseudomonas aeruginosa remains one of the most common nosocomial pathogens in intensive care units (ICUs). Although exogenous acquisition has been widely documented in outbreaks, its importance is unclear in non-epidemic situations. AIM: To elucidate the role of exogenous origin of P. aeruginosa in ICU patients. METHODS: A chronological analysis of the acquisition of P. aeruginosa was performed using samples collected in 2009 in the DYNAPYO cohort study, during which patients and tap water were screened weekly. Molecular relatedness of P. aeruginosa isolates was investigated by pulsed-field gel electrophoresis. Exogenous acquisition was defined as identification of a P. aeruginosa pulsotype previously isolated from another patient or tap water in the ICU. FINDINGS: The DYNAPYO cohort included 1808 patients (10,402 samples) and 233 water taps (4946 samples). Typing of 1515 isolates from 373 patients and 375 isolates from 81 tap water samples identified 296 pulsotypes. Analysis showed exogenous acquisition in 170 (45.6%) of 373 patients. The pulsotype identified had previously been isolated from another patient and from a tap water sample for 86 and 29 patients, respectively. The results differed according to the ICU. CONCLUSION: Exogenous acquisition of P. aeruginosa could be prevented in half of patients. The overall findings of this survey support the need for studies on routes of transmission and risk assessment approach to better define how to control exogenous acquisition in ICUs.

Prevalence, genetic diversity of and factors associated with ESBL-producing Enterobacterales carriage in residents of French nursing homes.

OBJECTIVE: To determine the prevalence and genotypic characteristics of extended-spectrum ß-lactamase-producing Enterobacterales (ESBLE) and carbapenemase-producing Enterobacterales (CPE) in nursing homes (NHs) in a French region. Risk factors associated with their carriage were also investigated. METHODS: A point-prevalence survey was proposed from November 2017 to June 2018 to NHs in the study region. Volunteer residents were screened for ESBLE and CPE carriage. Escherichia coli and Klebsiella pneumoniae isolates were genotyped using multi-locus sequence typing, pulsed-field gel electrophoresis (PFGE) and phylogrouping (for E. coli alone). Collective and individual data were analysed by random-effects logistic regression. RESULTS: The study was conducted in 18 NHs and included 262 patients. Fifty-two patients (19.8%) carried at least one ESBLE, corresponding to 56 isolates (42 E. coli, 11 K. pneumoniae and three others), while no CPE was detected. The majority (27/42) of ESBL E. coli belonged to phylogroup B2, and ST131 was over-represented in this subset (21/27). PFGE analysis revealed ST131 cross-transmission within NHs. Regarding ESBL K. pneumoniae, nine of 11 isolates belonged to ST663, and PFGE suggested diffusion of the clone in six NHs. Significant individual risk factors for colonization by ESBLE were: use of a shared bathroom, previous antibiotic use and recent history of hospitalization. Significant collective protective factors were proper compliance with glove use and support of the NH by a healthcare facility. CONCLUSION: This study shows that NHs in the study region are an important reservoir of ESBLE, whereas no residents were CPE carriers. The control of ESBLE in NHs should focus on antibiotic stewardship and excreta management policies.

[A case of periodic thyrotoxic paralysis]. / Le cas clinique du mois. Paralysie périodique thyrotoxique.

Periodic thyrotoxic paralysis (TPP) is a disease that usually affects Asian men from 20 to 50 years. It is secondary to genetic mutations of ion channels for the transport of sodium and potassium. It is characterized by recurrent and brutal bouts of paresis / flaccid paralysis, without sensory involvement, in a context of hyperthyroidism. Seizures preferentially affect proximal muscles and are often associated with severe hypokalemia. However, serious repercussions are rare, except in cases of extreme weakness of the respiratory muscles or cardiac arrhythmia induced by hypokalemia. The treatment of ionic disorders, followed by the correction of hyperthyroïdism, leads to the complete disappearance of symptoms.

La paralysie périodique thyrotoxique (PPT) est une maladie affectant généralement les hommes d'origine asiatique entre 20 et 50 ans. Elle est secondaire à des mutations génétiques de certains canaux ioniques permettant le transport du sodium et du potassium. Elle se caractérise par la survenue d'accès récurrents et brutaux de parésie/paralysie flasque, sans atteinte sensitive, dans un contexte d'hyperthyroïdie. Les crises touchent préférentiellement les muscles proximaux et sont très souvent associées à une hypokaliémie parfois sévère. Cette maladie n'a toutefois que rarement des répercussions graves, sauf en cas de faiblesse extrême des muscles respiratoires ou d'arythmie cardiaque induite par l'hypokaliémie. Le traitement des troubles ioniques, suivi de la correction de l'hyperthyroïdie, conduit à la disparition complète des symptômes.

Impact of a long-term antibiotic stewardship program targeting fluoroquinolones in a French local hospital.

INTRODUCTION: Fluoroquinolones (FQs) are major antibiotics but their wide use in hospital and community settings has led to an increased bacterial resistance against this antibiotic class. We aimed to assess the efficiency of an antibiotic stewardship program targeting FQs in a local hospital, and its impact on bacterial resistance. METHODS: This observational study was conducted in the local hospital of Morteau (Franche-Comté region, East of France). The hospital has 166 beds with health and medico-social sectors and a medical home affiliated with the facility. Local guidelines on empirical treatment regimens were released in 2007 aiming to reduce the use of FQs, especially for urinary tract infections. The following monitoring indicators were assessed: total consumption of antibiotics and of FQs (DDD/1,000 hospital patient-days), and resistance to nalidixic acid among Escherichia coli strains. Changes in the number of FQ packs sold in a community pharmacy were also recorded. RESULTS: The FQ consumption decreased by 85.6% between 2006 and 2015 (from 41.1 to 5.9 DDD/1,000 patient-days). The resistance to nalidixic acid among E. coli strains substantially decreased after remaining steady until 2011 (-57.2% between 2007 and 2015). The number of norfloxacin packs sold in the assessed community pharmacy decreased by 88%. CONCLUSION: Setting up an antibiotic stewardship program in a local hospital can lead to a substantial reduction in FQ use and in E. coli resistance to FQs. It may also have a positive impact on community prescriptions.

Antibiotics associated with acquisition of carbapenem-resistant Pseudomonas aeruginosa in ICUs: a multicentre nested case-case-control study.

Background: Carbapenem-resistant Pseudomonas aeruginosa (CRPA) strains are involved in severe infections, mostly in ICUs. Exposure to antibiotics other than carbapenems may be associated with isolation of CRPA; therefore, we aimed to identify those antibiotics using the case-case-control study design. Methods: A case-case-control study was conducted in 2015 in a prospective multicentre cohort that included 1808 adults hospitalized in 2009 in 10 French ICUs. Patients were screened for P. aeruginosa at admission to the ICU and then weekly. Cases were patients with CRPA and patients with carbapenem-susceptible P. aeruginosa (CSPA) isolation. Controls were patients without P. aeruginosa isolation, matched with each case according to centre, length of stay and hospitalization period. Effects of antibiotic exposure were explored, after adjusting for prior treatment with carbapenems and confounding factors comprising colonization pressure with two logistic regression models. The two models were compared to identify specific risk factors for CRPA isolation. Results: Fifty-nine CRPA, 83 CSPA and 142 controls were compared. In adjusted multivariable analyses, exposure to carbapenems and to antibiotics belonging to the group of ß-lactams inactive against P. aeruginosa were independent risk factors for CRPA isolation (OR, 1.205; 95% CI, 1.079-1.346 and OR, 1.101; 95% CI, 1.010-1.201, respectively). Conversely, exposure to ß-lactams active against P. aeruginosa was an independent protective factor for CSPA isolation (OR, 0.868; 95% CI, 0.772-0.976). Conclusions: Besides carbapenem exposure, exposure to ß-lactams inactive against P. aeruginosa was a specific risk factor for CRPA isolation. Clinicians should counterweigh the potential benefits of administering these antibiotics against the increased risk of CRPA infection.

[Subarachnoid hemorrhage resulting from a ruptured aneurysm : management in an emergency department]. / L'hémorragie méningée sur rupture d'anévrisme Prise en charge dans un service d'Urgences.

Subarachnoid hemorrhage is a neurovascular emergency affecting relatively young adults. A ruptured cerebral aneurysm is the main non-traumatic cause. Except for the non-specific symptom of headache, which frequently occurs in subarachnoid hemorrhage, the clinical presentation is atypical, making the diagnosis difficult with possible dramatic consequences. The diagnosis is based on the CT scan without contrast, which, when negative, will be followed by a lumbar puncture. The etiologic analysis is based on an urgent arteriography, revealing the characteristics of the aneurysm such as the anatomy of the intracerebral vessels, the identification of the bleeding site and the size and location of the aneurysm. The main complications are bleeding relapse, hydrocephaly and vasospasm. Early treatment is based on the final repair of the aneurysm through two possible approaches: microvascular neurosurgical clipping or coiling via endovascular access. This article discusses the diagnostic and therapeutic aspects of subarachnoid hemorrhage.

L'hémorragie méningée représente une urgence neuro-vasculaire grave affectant l'adulte relativement jeune. La rupture d'anévrisme cérébral en est la principale cause non traumatique. Hormis la présence fréquente de céphalées, son mode de présentation clinique est variable, ce qui rend le diagnostic difficile et peut entraîner des conséquences dramatiques. La mise au point diagnostique repose sur le scanner cérébral sans injection de produit de contraste qui, s'il est négatif, est suivi d'une ponction lombaire. Le bilan étiologique repose sur l'artériographie, laquelle renseigne sur les caractéristiques de l'anévrisme : anatomie des vaisseaux intracérébraux, identification du site de saignement, taille et localisation de l'anévrisme. Les principales complications sont la récidive de saignement, l'hydrocéphalie et le vasospasme. Le traitement, précoce, repose sur la réparation définitive de l'anévrisme par deux principaux abords : la voie neurochirurgicale (clippage microvasculaire) ou le traitement endovasculaire par pose de spires métalliques (ou coiling). Cet article aborde les aspects diagnostiques et thérapeutiques de la prise en charge de l'hémorragie méningée.

Global emergence of the widespread Pseudomonas aeruginosa ST235 clone.

OBJECTIVES: Despite the non-clonal epidemic population structure of Pseudomonas aeruginosa, several multi-locus sequence types are distributed worldwide and are frequently associated with epidemics where multidrug resistance confounds treatment. ST235 is the most prevalent of these widespread clones. In this study we aimed to understand the origin of ST235 and the molecular basis for its success. METHODS: The genomes of 79 P. aeruginosa ST235 isolates collected worldwide over a 27-year period were examined. A phylogenetic network was built, using a Bayesian approach to find the Most Recent Common Ancestor, and we identified antibiotic resistance determinants and ST235-specific genes. RESULTS: Our data suggested that the ST235 sublineage emerged in Europe around 1984, coinciding with the introduction of fluoroquinolones as an antipseudomonal treatment. The ST235 sublineage seemingly spread from Europe via two independent clones. ST235 isolates then appeared to acquire resistance determinants to aminoglycosides, ß-lactams and carbapenems locally. Additionally, we found that all the ST235 genomes contained the exoU-encoded exotoxin and identified 22 ST235-specific genes clustering in blocks and implicated in transmembrane efflux, DNA processing and bacterial transformation. These unique combinations of genes may have contributed to the poor outcome associated with P. aeruginosa ST235 infections and increased the ability of this international clone to acquire mobile resistance elements. CONCLUSION: Our data suggest that P. aeruginosa ST235 (a) has become prevalent across the globe potentially due to the selective pressure of fluoroquinolones and (b) readily became resistant to aminoglycosides, ß-lactams and carbapenems through mutation and acquisition of resistance elements among local populations.

[How I treat a poisoning with tricyclic antidepressants : potential role of a treatment with lipid emulsion]. / Comment je traite l'intoxication aux antidépresseurs tricycliques : efficacité potentielle d'un traitement par émulsion lipidique.

Poisoning with tricyclic antidepressants is common and can be life-threatening. The classic management is well known (chelating gastrointestinal, sodium bicarbonate, benzodiazepine, norepinephrine). A few years ago, a treatment with lipid emulsion, previously used in local anesthetics poisoning, has been successfully tested in tricyclic poisoning with cardiac arrest. We are currently unable to explain the exact mechanism of this treatment but it could have a place in the treatment of severe tricyclic poisoning with hemodynamic instability in addition to the conventional treatment.

L'intoxication aux antidépresseurs tricycliques n'est pas rare et peut engager le pronostic vital du patient. La prise en charge classique est bien connue (chélateurs gastrointestinaux, bicarbonate de sodium, benzodiazépines, amines vasoactives). Il y a quelques années, un traitement par émulsion lipidique, jusque-là utilisé dans les intoxications aux anesthésiques locaux, a été testé avec succès dans les intoxications aux tricycliques en arrêt cardio-respiratoire. Nous sommes, actuellement, incapables d'expliquer le fonctionnement exact de ce traitement, mais il pourrait avoir une place dans la prise en charge des intoxications aux tricycliques avec instabilité hémodynamique échappant au traitement conventionnel.

[Mediastinitis on oesphagal perforation]. / Le cas clinique du mois. Médiastinite sur perforation oesophagienne.

Vomiting is less benign than it seems … Esophageal perforations and ruptures cause the contents of the esophageal lumen to leak into the cervical retroesophageal space, the mediastinum or the abdomen, which can trigger, among other adverse events, a mediastinitis, the diagnosis of which is difficult and should be established as promptly as possible. It's a rare pathology involving a high risk of diagnostic errors.

Les perforations et les ruptures de l'œsophage mettent en communication la lumière oesophagienne avec les espaces rétro-oesophagiens cervicaux, le médiastin ou l'abdomen. Ceci peut, entre autres, provoquer une médiastinite, dont le diagnostic, difficile, doit être le plus précoce possible. Il s'agit d'une pathologie rare entraînant des risques importants d'erreur diagnostique.

Association between Pseudomonas aeruginosa positive water samples and healthcare-associated cases: nine-year study at one university hospital.

OBJECTIVE: To study the association between the results of water samples and Pseudomonas aeruginosa healthcare-associated cases in a French university hospital. METHODS: Generalized Estimating Equations were used on complete case and imputed datasets. The spatial unit was the building and the time unit was the quarter. RESULTS: For the period 2004-2013, 2932 water samples were studied; 17% were positive for P. aeruginosa. A higher incidence of P. aeruginosa cases was associated with a higher proportion of positive water samples (P=0.056 in complete case analysis and P=0.031 with the imputed dataset). The association was no longer observed when haematology and intensive care units were excluded, but was significant in analyses of data concerning intensive care units alone (P<0.001). CONCLUSION: This study suggests that water outlet contamination in hospitals can lead to an increase in healthcare-associated P. aeruginosa cases in wards dealing with susceptible patients, but does not play a significant role in other wards.

Detection of temporal clusters of health care-associated infections or colonizations with Pseudomonas aeruginosa.

We investigated temporal clusters of Pseudomonas aeruginosa cases between 2005 and 2014 in 1 French university hospital, overall and by ward, using the Kulldorff method. Clusters of positive water samples were also investigated at the whole hospital level. Our results suggest that water outlets are not closely involved in the occurrence of clusters of P aeruginosa cases.

mcr-1 is borne by highly diverse Escherichia coli isolates since 2004 in food-producing animals in Europe.

OBJECTIVES: In November 2015, a plasmid-mediated colistin resistance, MCR-1, was described in animals, food and humans in China, and it was considered as a potential emerging threat to public health. Therefore, we screened for the mcr-1 gene a European collection of colistin-resistant Escherichia coli (n=218) and Salmonella spp. (n=74) isolated from diseased food-producing animals between 2004 and 2014 and characterized the mcr-1-positive clones. METHODS: Screening for mcr-1 gene was performed by PCR on isolates for which inhibition diameter was <15 mm around a 50 µg disk of colistin. Positive E. coli isolates were then characterized by phylogrouping, multilocus sequence typing and pulsed-field gel electrophoresis typing. Antibiotic susceptibility was determined by disk diffusion testing or by broth microdilution. RESULTS: Among the collection, 42 E. coli and three Salmonella spp. were positive for mcr-1, with continuous detection since 2004 mainly from bovine and swine digestive infections. Most of the mcr-1-positive strains were resistant to amoxicillin and cotrimoxazole but remained susceptible to cephalosporins, carbapenems and piperacillin/tazobactam. All but one isolate were resistant to colistin, with a minimum inhibitory concentration of >2 mg/L. Most of the mcr-1-positive E. coli belonged to the phylogroup A with two prevalent clonal complexes, CC10 and CC165, in which sequence type 10 and sequence type 100 were overrepresented and pulsed-field gel electrophoresis typing revealed a high diversity of pulsotypes. CONCLUSIONS: MCR-1 was detected yearly in European food-producing animal since 2004 with a high diversity of pulsotypes supporting the dissemination of mcr-1 via plasmids.