BACKGROUND:Staphylococcus haemolyticus is a clinically relevant coagulase-negative staphylococcus frequently responsible for hospital-acquired infections, especially in premature newborns. AIM:To describe an outbreak of multidrug-resistant S. haemolyticus in a neonatal department. METHODS:The outbreak was investigated using classical methods, including screening of the patients, genotyping of the isolates and environmental survey. Numerous infection control measures were implemented. FINDINGS:In 2022 and 2023, a clonal outbreak of multidrug-resistant S. haemolyticus ST29 (40 infections and 71 carriages) occurred in the neonatology department of a University Hospital in France. The infection control measures implemented only partially controlled the outbreak. Although our investigation did not clearly identify the source and mode of transmission, a reservoir constituted by patients and transmission by healthcare workers are the most likely. This episode occurred in a context of countrywide outbreaks of S. haemolyticus ST29 in several French neonatology departments. CONCLUSION:This prolonged outbreak of S. haemolyticus ST29 accounted for the increase in the incidence of S. haemolyticus-related infections in French neonatology departments. Implementation of proactive measures is crucial to limit the spread of such pathogens in neonatal ICUs.

OBJECTIVES:We assessed the contamination with Legionella pneumophila (Lp) of the hot water network (HWN) of a hospital, mapped the risk of contamination, and evaluated the relatedness of isolates. We further validated phenotypically the biological features that could account for the contamination of the network. METHODS:We collected 360 water samples from October 2017 to September 2018 in 36 sampling points of a HWN of a building from a hospital in France. Lp were quantified and identified with culture-based methods and serotyping. Lp concentrations were correlated with water temperature, date and location of isolation. Lp isolates were genotyped by pulsed-field gel electrophoresis and compared to a collection of isolates retrieved in the same HWN two years later, or in other HWN from the same hospital. RESULTS:207/360 (57.5%) samples were positive with Lp. In the hot water production system, Lp concentration was negatively associated with water temperature. In the distribution system, the risk of recovering Lp decreased when temperature was >55 °C (p < 10-3), the proportion of samples with Lp increased with distance from the production network (p < 10-3), and the risk of finding high loads of Lp increased 7.96 times in summer (p = 0.001). All Lp isolates (n = 135) were of serotype 3, and 134 (99.3%) shared the same pulsotype which is found two years later (Lp G). In vitro competition experiments showed that a 3-day culture of Lp G on agar inhibited the growth of a different pulsotype of Lp (Lp O) contaminating another HWN of the same hospital (p = 0.050). We also found that only Lp G survived to a 24h-incubation in water at 55 °C (p = 0.014). CONCLUSION:We report here a persistent contamination with Lp of a hospital HWN. Lp concentrations were correlated with water temperature, season, and distance from the production system. Such persistent contamination could be due to biotic parameters such as intra-Legionella inhibition and tolerance to high temperature, but also to the non-optimal configuration of the HWN that prevented the maintenance of high temperature and optimal water circulation.

The worldwide spread of Klebsiella pneumoniae producing extended-spectrum β-lactamase (ESBL-Kp) is a significant threat. Specifically, various pandemic clones of ESBL-Kp are involved in hospital outbreaks and caused serious infections. In that context, we assessed the phenotypic and molecular features of a collection of ESBL-Kp isolates in a French university hospital and evaluated the occurrence of potential cross-transmissions. Over a 2-year period (2017–2018), 204 non-duplicate isolates of ESBL-Kp were isolated from clinical (n = 118, 57.8%) or screening (n = 86, 42.2%) sample cultures. These isolates were predominantly resistant to cotrimoxazole (88.8%) and ofloxacin (82.8%) but remained susceptible to imipenem (99.3%) and amikacin (93.8%). CTX-M-15 was the most frequent ESBL identified (83.6%). Multilocus sequence typing and pulse-field gel electrophoresis analysis showed an important genetic variability with 41 sequence types (ST) and 50 pulsotypes identified, and the over representation of the international epidemic clones ST307 and ST405. An epidemiological link attesting probable cross-transmission has been identified for 16 patients clustered in 4 groups during the study period. In conclusion, we showed here the dissemination of pandemic clones of ESBL-Kp in our hospital on a background of clonal diversity.

The worldwide spread of E. coli ST131 has significantly contributed to the dissemination of E. coli producing extended-spectrum β-lactamases (ESBL). In a French University hospital, we assessed the molecular features of ESBL-producing E. coli and identified risk factors in patients for colonization or infection with E. coli ST131. Over a 2-year period (2015–2017), each patient with at least one clinical isolate or one screening isolate positive with ESBL-producing E. coli were included (n = 491). The ST131 clonal group accounted for 17.5% (n = 86) of all ESBL-producing E. coli and represented 57.3% isolates of phylogroup B2. FimH -based sub-typing showed that 79.1% (68/86) of ST131 isolates were fimH30, among which 67.6% (n = 46), 20.6% (n = 14) and 11.8% (n = 8) isolates harbored genes encoding the ESBL CTX-M-15, CTX-M-27, and CTX-M-14, respectively. The multivariate analysis identified two factors independently associated with ST131 ESBL-producing E. coli isolates: infection (Odds ratio [OR] = 1.887, 95% confidence interval [CI]: 1.143–3.115; p = 0.013) and community acquisition (OR = 2.220, 95% CI: 1.335–3.693; p = 0.002). In conclusion, our study confirmed the predominance of ST131 clonal group among ESBL-producing E. coli and the difficulty to identify common risk factors associated with carriage of this pandemic clonal group.

Background. The issue of contact precautions as contributory factors for reducing Pseudomonas aeruginosa (Pa) infections in intensive care units (ICUs) remains questioned. We evaluated the impact of the addition of contact precautions to standard precautions for Pa-positive patients on incidence of ICU-acquired Pa infections. Methods. In this multicenter, cluster-randomized crossover trial, 10 French ICUs were randomly assigned (1:1) to sequence 0-1 (6-month control period [CP]/3-month washout period/6-month intervention period [IP]) or sequence 1-0 (6-month IP/3-month washout period/6-month CP). A surveillance screening program for Pa was implemented. Competing-risks regression models were built with death and discharge without the occurrence of ICU-acquired Pa infection (the primary outcome) as competing events. Models were adjusted for within-ICU correlation and patient- and ICU-level covariates. The Simpson diversity index (SDI) and transmission index (TI) of Pa isolates were derived from pulsed-field gel electrophoresis typing. Results. Within recruited ICUs, the cumulative incidence and incidence rate of ICU-acquired Pa infections were 3.38% (55/1625) versus 3.44% (57/1658) and 3.31 versus 3.52 per 1000 patient-days at risk during the CP and IP, respectively. Multivariable models indicated that the intervention did not significantly change the cumulative incidence (subdistribution hazard ratio,.91; 95% confidence interval [CI], .49-1.67; P =.76) or rate (cause-specific hazard ratio, 1.36; 95% CI,.71-2.63; P =.36) of the primary outcome. SDI and TI did not significantly differ between CP and IP. Conclusions. The addition of contact precautions to standard precautions for Pa-positive patients with a surveillance screening program does not significantly reduce ICU-acquired Pa infections in non-outbreak situations.

Background The spread of carbapenemase-producing Enterobacteriaceae (CPE) in the Southwest Indian Ocean area (SIOA) is poorly documented. Reunion Island is a French overseas territory located close to Madagascar and connected with Southern Africa, Indian sub-continent and Europe, with several weekly flights. Here we report the results of the CPE surveillance program in Reunion Island over a six-year period. Methods All CPE were collected between January 2011 and December 2016. Demographics and clinical data of the carrier patients were collected. We determined their susceptibility to antimicrobials, identified the carbapenemases and ESBL by PCR and sequencing, and explored their genetic relationship using pulsed-field gel electrophoresis and multi-locus sequence typing. Results A total of 61 CPEs isolated from 53 patients were retrieved in 6 public or private laboratories of the island. We found that 69.8% of CPE patients were linked to a foreign country of SIOA and that almost half of CPE cases (47.2%) reached the island through a medical evacuation. The annual number of CPE cases strongly increased over the studied period (one case in 2011 vs. 21 cases in 2016). A proportion of 17.5% of CPE isolates were non-susceptible to colistin. bla(NDM) was the most frequent carbapenemase (79.4%), followed by bla(IMI) (11.1%), and bla(IMP-10) (4.8%). Autochtonous CPE cases (30.2%) harboured CPE isolates belonging to a polyclonal population. Conclusions Because the hospital of Reunion Island is the only reference healthcare setting of the SIOA, we can reasonably estimate that its CPE epidemiology reflects that of this area. Mauritius was the main provider of foreign CPE cases (35.5%). We also showed that autochthonous isolates of CPEs are mostly polyclonal, thus unrelated to cross-transmission. This demonstrates the local spread of carbapenemase-encoding genes (i.e. bla(NDM)) in a polyclonal bacterial population and raises fears that Reunion Island could contribute to the influx of NDM-carbapenemase producers into the French mainland territory.

The aim of the study was to determine factors associated with spread of linezolid (LNZ)-resistant Staphylococcus epidermidis isolates in a surgical intensive care unit (ICU). A case-control study was conducted in one French adult surgical ICU. From January 2012 to December 2016, patients with at least a single positive LNZ-resistant S. epidermidis blood culture were matched to control with LNZ-susceptible S. epidermidis blood culture in a 1:4 manner. Cases were compared to controls regarding baseline clinical characteristics and LNZ exposure before positive blood culture. Bacterial isolates were genotyped by using pulsed-field gel electrophoresis (PFGE) and MLST. We identified 13 LNZ-resistant S. epidermidis isolates, 1 in 2012, 3 in 2014, 6 in 2015, and 3 in 2016. LNZ use increased steadily from 8 DDDs/100 patient days in 2010 to 19 in 2013 and further decrease by more of 50% in 2015 and 2016. The only independent risk factors associated to LNZ-resistant S. epidermidis isolation were length of stay in ICU before infection (OR 1.45; 95% CI 1.07–1.98), prior exposure to LNZ (OR 109; 95% CI 3.9–3034), and Charlson comorbidities score (OR 3.19; 95% CI 1.11–9.14). PFGE typing showed that all LNZ-resistant isolates were clonal belonging to ST2 and that LNZ-susceptible isolates were highly diverse. We report herein that previous exposure to LNZ substantially increased the risk of occurrence of LNZ resistance in S. epidermidis even in the case of clonal spread of LNZ-resistant isolates. These findings highlight the need for reducing the use of LNZ to preserve its efficacy in the future.

The typing of epidemic bacterial pathogens in hospitals relies on DNA-based, expensive, and time-consuming techniques, that are often limited to retrospective studies. However, the quick identification of epidemic pathogens in the routine of the microbiology laboratories would expedite infection control procedures that limit the contamination of new patients. IR Biotyper (Bruker Daltonics GmbH) is a new typing machine based on Fourier-transform infrared (FTIR) spectroscopy which generates spectra, aiming at typing the micro-organisms within 3 h. This technique discriminates the isolates by exploring the differences of the surface cell polysaccharides. In this work, we evaluated the ability of the FTIR spectroscopy to recognize Gram-negative bacilli clones responsible for hospital outbreaks. Isolates of Pseudomonas aeruginosa (n = 100), Klebsiella pneumoniae (n = 16), Enterobacter cloacae (n = 23), and Acinetobacter baumannii (n = 20) were typed by the reference methods Multi-Locus Sequence Typing (defining sequence types - STs) along with or without pulsed field gel electrophoresis (PFGE) (defining pulsotypes), and by FTIR spectroscopy. The congruence of FTIR spectroscopy clustering was compared to those of MLST and PFGE by Adjusted Rand index and Adjusted Wallace coefficient. We found that FTIR spectroscopy accurately clustered P. aeruginosa, K. pneumoniae, and E. cloacae isolates belonging to the same ST. The performance of the FTIR spectroscopy was slightly lower for A. baumannii. Furthermore, FTIR spectroscopy also correctly clustered P. aeruginosa isolates having a similar pulsotype. Overall, the IR Biotyper can quickly (in less than 3 h) detect the spread of clones of P. aeruginosa, K. pneumoniae, E. cloacae, and A. baumannii. The use of this technique by clinical microbiology laboratories may help to tackle the spread of epidemic clones by the quick implementation of infection control measures.

Introduction: Pseudomonas aeruginosa is one of the most common nosocomial pathogens, known with a wide resistance to antimicrobials. Carbapenemases producing Pseudomonas aeruginosa is a growing global public health concern as this pathogen is easily transmissible among patients. Metallo-Beta-lactamases is the most important class of these carbapenemases with their broad-spectrum resistance profile. This study was conducted to investigate the prevalence of MBL-producing P. aeruginosa collected in an Algerian hospital. Methodology: All Metallo-β-lactamase (MBL)-producing P. aeruginosa isolates recovered from patients during a 2 years period (2015-2016) were studied using a combination of phenotypic and molecular typing methods (susceptibility testing, molecular characterization of carbapenemase-encoding genes, multi-locus sequence typing and pulsed-field gel electrophoresis). Results: A total of twenty-six MBL producing P. aeruginosa of 188 isolates were investigated. The burns unit ranked in the first position of the majority of identified cases with 73.07%. About 73.07% of total MBL isolates were mainly isolated from pus samples. The studied isolates were subjected to the molecular typing, in which 4 different Dra1-PFGE patterns and 3 sequences type were assigned (ST244, ST381, and ST1076), and all isolates were revealed positive for VIM-4. Conclusions: We report the third description of blaVIM-4 in Algeria indicating the emergence and spread of carbapenemase-encoding genes among P. aeruginosa in the hospital environment.

Although Pseudomonas aeruginosa has a non-clonal epidemic population structure, recent studies have provided evidence of the existence of epidemic high-risk clones. The aim of this study was to assess the molecular epidemiology of P. aeruginosa isolates responsible for infections in French ICUs, regardless of resistance patterns. For a 1-year period, all non-duplicate P. aeruginosa isolated from bacteremia and pulmonary infections in ten adult ICUs of six French university hospitals were characterized by antimicrobial susceptibility testing and genotyping (MLST and PFGE). We identified β-lactamases with an extended spectrum phenotypically and by sequencing. The 104 isolates tested were distributed in 46 STs, of which 7 epidemic high-risk (EHR) clones over-represented: ST111, ST175, ST235, ST244, ST253, ST308, and ST395. Multidrug-resistant (MDR) isolates mostly clustered in these EHR clones, which frequently spread within hospitals. Only one ST233 isolate produced the carbapenemase VIM-2. PFGE analysis suggests frequent intra-hospital cross-transmission involving EHR clones. For ST395 and ST308, we also observed the progression from wild-type to MDR resistance pattern within the same PFGE pattern. Molecular epidemiology of P. aeruginosa in French ICUs is characterized by high clonal diversity notably among antimicrobial susceptible isolates and the over-representation of EHR clones, particularly within MDR isolates, even though multidrug resistance is not a constant inherent trait of EHR clones.

Objective: The aim of this study was to investigate the resistance mechanisms of carbapenem-resistant Enterobacteriaceae clinical strains recovered from Al Thawra University Hospital, Sana'a, Yemen. Methods: A total of 27 isolates showing decreased susceptibility to carbapenems were obtained from different clinical specimens in Al Thawra Hospital, Sana'a, Yemen. Strains were identified by Matrix Assisted Laser Desorption Ionization Time-Of-Flight spectroscopy. Susceptibility to antibiotics was determined by the disk diffusion method on Mueller Hinton agar. Carbapenemases-encoding genes, extended-spectrum β-lactamases (ESBLs), and plasmid-mediated quinolone resistance (PMQR) genes were screened by PCR. Bacterial isolates were typed by multilocus sequence typing (MLST). Results: Carbapenemase genes detection and sequencing showed that 18 (66.7%) isolates were Klebsiella pneumoniae (NDM-1, n = 13; NDM-1 + OXA-48, n = 3; OXA-48, n = 1; OXA-232, n = 1), 6 (22.2%) were Escherichia coli (NDM-5, n = 3; OXA-181, n = 2; OXA-48, n = 1), and 3 (11.1%) were Enterobacter cloacae (NDM-1, n = 1; OXA-181, n = 2). In addition the ESBL gene blaCTX-M-15 was detected in 14 K. pneumoniae and 2 E. coli isolates, and the blaCTX-M-216 was found in 1 E. coli isolate. Fifteen isolates were PMQR positive including qnrB1 (n = 1), qnrS1 (n = 5), qnrS4 (n = 2), and aac-(6')-Ib-cr (n = 7). The MLST typing showed a diversity of sequence type (ST) clones including Escherichia coli ST410 (3), ST448 (2), and ST648; Enterobacter cloacae ST78 and ST270; and Klebsiella pneumoniae ST395 (2), ST309, ST23, ST35, ST1728, ST15, ST231, and ST1428. Conclusion: This study reports the first description of OXA-48-like-producing Enterobacteriaceae and NDM-5 enzymes in E. coli in Yemen.

The epidemiology of Staphylococcus aureus is changing and several surveillances worldwide have evidenced an increasing incidence of S. aureus bloodstream infections (BSIs). Here, we described the long-term epidemiology of the emergent clonal group CC398 among S. aureus isolated from BSIs in our French university hospital between 2010 and 2017. Each patient with at least one blood culture positive with S. aureus during the study period was included (N = 1455). Cefoxitin susceptibility was determined using the disk diffusion method according to EUCAST recommendations. CC398 isolates were first screened from the whole S. aureus collection with a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) typing method confirmed by a CC398-specific PCR. In our hospital, the incidence of hospital- and community-acquired BSIs due to S. aureus and MSSA increased in parallel between 2010 and 2017 while that of BSIs with MRSA decreased. The prevalence of CC398 isolates among S. aureus from BSIs increased from 3.6 in 2010 to 20.2% in 2017 (p < 0.05). CC398-MRSA emerged but remains very sparse. Our data suggested that CC398-MSSA disseminates in the community. We showed here the emergence and the diffusion of CC398-MSSA, a subclone associated with invasive infections, in our hospital. The monitoring of this particular human-adapted S. aureus clone is needed and genomic studies will have to identify the determinants of its diffusion.

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The spread of carbapenemase-producing Enterobacteriaceae in the Southwest Indian Ocean islands is poorly known. Here we describe an outbreak of colistin-resistant Enterobacter cloacae harbouring blaIMI-1 in the French overseas department of Mayotte. Between October 2015 and January 2017, all isolates of imipenem-non-susceptible E. cloacae at Mayotte Medical Center and University Hospital of Reunion Island were screened for carbapenemase production. Positive isolates were typed by pulsed-field gel electrophoresis and whole-genome sequencing (WGS)-based multilocus sequence typing (MLST), and all β-lactamase genes were identified by PCR and sequencing. Resistance profiles were determined by agar diffusion and Etest. Genetic support of the blaIMI-1 gene was determined by WGS. A total of 18 E. cloacae isolates harbouring blaIMI-1 were detected in 17 patients from Mayotte. Pulsed-field gel electrophoresis (PFGE) analysis showed 16 of the 18 strains to be clonally related and belonging to ST820. Based on clinical data, this outbreak most likely had a community origin. The blaIMI-1 gene in the 18 isolates was carried by a new variant of an integrative mobile element involving the Xer recombinases, called EcloIMEX-8. The mcr-1–mcr-5 genes were absent from the collection. The isolates belonged to E. cloacae cluster XI, known to be colistin heteroresistant. Here we report the first outbreak of IMI-1-producing Enterobacteriaceae. IMI-1-producers may be underdetected in microbiology laboratories because of their unusual antimicrobial resistance profile (resistant to imipenem but with intermediate resistance to ertapenem and susceptible to extended-spectrum cephalosporins) and the absence of blaIMI-1 in the panel of genes targeted by molecular diagnostic kits.

OBJECTIVE To describe an outbreak of multidrug-resistant Pseudomonas aeruginosa in which the hospital waste-pipe system was the likely source of contamination and to report the bundle of measures that facilitated the long-term control of the outbreak. DESIGN Outbreak investigation. SETTING The hematology unit of a tertiary-care referral center. PATIENTS Patients who were colonized or infected with P. aeruginosa belonging to the clonal outbreak. METHODS Patients admitted to our 15-bed stem-cell transplantation hematology unit were screened for P. aeruginosa carriage. Pseudomonas aeruginosa isolates were also obtained from diagnostic samples. We assessed the microbiological contamination of P-traps, water and toilets for 42 months. Extended-spectrum β-lactamases (ESBLs) and metallo-β-lactamases (MBLs) were screened and identified by polymerase chain reaction (PCR) and sequencing. Molecular typing of ESBL- or MBL-producing isolates was carried out using pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). RESULTS From 2009 to 2013, a biclonal outbreak of IMP-19–producing ST235 (11 cases) and IMP-29–producing ST111 (10 cases) of P. aeruginosa occurred. The environmental investigation strongly suggested that P-traps were the reservoirs for the outbreak strains. A bundle of infection control measures, including engineering interventions on water outlets and disinfection of P-traps, controlled the outbreak. CONCLUSIONS We report a prolonged outbreak of IMP-producing high-risk clones of P. aeruginosa , for which P-traps seems to play a major role in cross-transmission. It appears essential to implement proactive measures to limit the bacterial load in water fittings of high-risk units. Infect Control Hosp Epidemiol 2018;39:164–169

The presence of P. aeruginosa in water supply is clearly identified as a risk factor for P. aeruginosa infection in critical care units, even if routes of transmission are often unclear and remain a matter of debate. We determined here the frequency of U-bends contaminated with P. aeruginosa in high-risk units and described the population structure of this opportunistic pathogen in a non-outbreak situation. Eighty-seven U-bends from sinks of rooms in five wards were sampled 3 times and P. aeruginosa was detected in 121 of the 261 (46.4%) U-bend samples. We genotyped 123 P. aeruginosa isolates with pulsed-field gel electrophoresis and multilocus sequence typing and found 41 pulsotypes distributed in 21 Sequence Types (STs). Seven major ST (ST111, CC235, CC253, ST520, ST539, ST1216, and ST1725) were overrepresented in the collection, including the high-risk clones ST111, CC253, and CC235. The distribution of the 21 STs in the cladogram of the species was uneven with most major STs clustering into 2 clades. The major STs were found in different units and buildings and could be represented by a high diversity of pulsotypes. Altogether, this suggests a long term presence of P. aeruginosa in the hospital water network, possibly contaminated by the distribution water or by plumbing fittings before putting into service. Analysis of resistance rates showed that the deficiency of porin OprD was very frequent in U-bends isolates that may benefit from this resistance mechanism in hospital water fittings. In conclusion, our study demonstrates that U-bends of high-risk units are very frequently contaminated with P. aeruginosa with a moderate genomic diversity and with an over-representation of adapted clones.

Sequence type 131 (ST131) is a predominant lineage among extraintestinal pathogenic Escherichia coli. It plays a major role in the worldwide dissemination of E. coli producing extended-spectrum beta-lactamases (ESBLs). Here we describe the long-term epidemiology of this clonal group in a French university hospital, where the incidence of ESBL-producing E. coli has increased from 0.018 case per 1000 patient-days in the year 2000 to 0.50 case per 1000 patient-days in 2014. The first of the 141 ST131 isolates was recovered in 2006, and the ST131 clonal group accounted for 18.1% of total ESBL-producing E. coli over the whole period (2000-2014). Subclonal typing showed that 75.9% (107/141) of ST131 isolates were H30, of which 81.3% (87/107) were H30-Rx. The large majority (137/141) of ESBLs produced were of the CTX-M group, with 94 CTX-M-15, 19 CTX-M-1, 10 CTX-M-27, 8 CTX-M-14 and four other CTX-M types (n = 6). Pulsed-field gel electrophoresis (PFGE) analysis showed high diversity, which increased during the course of the study. The 141 ST131 isolates clustered in 53 pulsotypes (PTs), with 2 dominant PTs (PT14 and PT13) with 36 and 17 isolates, respectively. These findings showed that ST131 was a predominant clone among ESBL-producing E. coli in our hospital, even though it only accounted for <20%. Moreover, ST131 should be regarded not as a unified entity but as a cluster of distinct clonal subsets even if the increase in resistance within ST131 has a strong clonal basis, being attributable mainly to the spread of C1/H30-R and C2/H30-Rx clades. (C) 2016 Elsevier B.V. and International Society of Chemotherapy. All rights reserved.

Reliable molecular typing methods are necessary to investigate the epidemiology of bacterial pathogens. Reference methods such as multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE) are costly and time consuming. Here, we compared our newly developed double-locus sequence typing (DLST) method for Pseudomonas aeruginosa to MLST and PFGE on a collection of 281 isolates. DLST was as discriminatory as MLST and was able to recognize "high-risk" epidemic clones. Both methods were highly congruent. Not surprisingly, a higher discriminatory power was observed with PFGE. In conclusion, being a simple method (single-strand sequencing of only 2 loci), DLST is valuable as a first-line typing tool for epidemiological investigations of P. aeruginosa. Coupled to a more discriminant method like PFGE or whole genome sequencing, it might represent an efficient typing strategy to investigate or prevent outbreaks.

Background Pseudomonas aeruginosa is a major human pathogen, which also affects animals. It is thought that P. aeruginosa has a non-clonal epidemic population structure, with distinct isolates found in humans, animals or the environment. However, very little is known about the structure of the P. aeruginosa population from diseased animals. Data on antimicrobial resistance are also scarce. Results Thirty-four already registered and 19 new MLST profiles were identified. Interestingly, a few clones were more prevalent, and clones associated to human outbreaks were also detected. Multidrug resistance phenotypes were overall rare. Conclusion We highlight the non clonal structure of the population and show a higher prevalence of specific clones, possibly correlating with higher pathogenicity. The low proportion of antimicrobial resistance contrasts with the high resistance rate of human isolates.

Fluoroquinolone (FQ) agents are a potential resort to treat infection due to Enterobacteriaceae producing extended spectrum β-lactamase and susceptible to FQ. In a context of increase of non-susceptibility to carbapenems among Enterobacteriaceae, we characterized FQ resistance mechanisms in 75 Enterobacter cloacae isolates non-susceptible to ertapenem in North-Eastern France in 2012 and describe the population structure by pulsed field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST). Among them, 14.7% (12/75) carried a carbapenemase-encoding gene. Except one isolate producing VIM-1, the carbapenemase-producing isolates carried the well-known IncL/M pOXA48a plasmid. Most of the isolates (59/75) harbored at least a FQ-R determinant. qnr genes were predominant (40%, 30/75). The MLST study revealed that E. cloacae isolates' clonality was wide [24 different sequence types (STs)]. The more widespread STs were ST74, ST101, ST110, ST114, and ST133. Carbapenem MICs were higher for E. cloacae ST74 than for other E. cloacae isolates. Plasmid-mediated quinolone resistance determinants were more often observed in E. cloacae ST74 isolates. These findings showed that (i) pOXA-48a is spreading in North-Eastern France, (ii) qnr is preponderant in E. cloacae, (iii) E. cloacae comprised a large amount of lineages spreading in North-Eastern France, and (iv) FQ as an alternative to β-lactams to treat ertapenem non-susceptible Enterobacteriaceae are compromised.