Epidemiological Description and Detection of Antimicrobial Resistance in Various Aquatic Sites in Marseille, France.

Antibiotic resistance is a worldwide public health concern and has been associated with reports of elevated mortality. According to the One Health concept, antibiotic resistance genes are transferrable to organisms, and organisms are shared among humans, animals, and the environment. Consequently, aquatic environments are a possible reservoir of bacteria harboring antibiotic resistance genes. In our study, we screened water and wastewater samples for antibiotic resistance genes by culturing samples on different types of agar media. Then, we performed real-time PCR to detect the presence of genes conferring resistance to beta lactams and colistin, followed by standard PCR and gene sequencing for verification. We mainly isolated Enterobacteriaceae from all samples. In water samples, 36 Gram-negative bacterial strains were isolated and identified. We found three extended-spectrum ß-lactamase (ESBL)-producing bacteria-Escherichia coli and Enterobacter cloacae strains-harboring the CTX-M and TEM groups. In wastewater samples, we isolated 114 Gram-negative bacterial strains, mainly E. coli, Klebsiella pneumoniae, Citrobacter freundii and Proteus mirabilis strains. Forty-two bacterial strains were ESBL-producing bacteria, and they harbored at least one gene belonging to the CTX-M, SHV, and TEM groups. We also detected carbapenem-resistant genes, including NDM, KPC, and OXA-48, in four isolates of E. coli. This short epidemiological study allowed us to identify new antibiotic resistance genes present in bacterial strains isolated from water in Marseille. This type of surveillance shows the importance of tracking bacterial resistance in aquatic environments. IMPORTANCE Antibiotic-resistant bacteria are involved in serious infections in humans. The dissemination of these bacteria in water, which is in close contact with human activities, is a serious problem, especially under the concept of One Health. This study was done to survey and localize the circulation of bacterial strains, along with their antibiotic resistance genes, in the aquatic environment in Marseille, France. The importance of this study is to monitor the frequency of these circulating bacteria by creating and surveying water treatments.

The Effect of Decreased Antipseudomonal Drug Consumption on Pseudomonas aeruginosa Incidence and Antimicrobial Susceptibility Profiles over 9 Years in a Lebanese Tertiary Care Center.

Pseudomonas aeruginosa (PAE) is intrinsically resistant to numerous classes of antimicrobials such as tetracycline and ß-lactam antibiotics. More epidemiological surveillance studies on the antimicrobial susceptibility profiles of PAE are needed to generate clinically significant data and better guided therapeutic options. We describe and analyze in a retrospective study the epidemiologic trends of 1827 Pseudomonas spp. isolates (83.5% PAE, 16.4% Pseudomonas sp., and 0.2% Pseudomonas putida) from various clinical specimens with their resistance patterns to antimicrobial consumption at a tertiary medical center in Lebanon between January 2010 and December 2018. We report a significant drop in the incidence of PAE from sputum (p-value = 0.05), whereas bloodstream infection isolation density showed no trend over the study period. We also registered a minimal but statistically significant drop in resistance of Pseudomonas to certain antibiotics and a decrease in the consumption of antipseudomonal antibiotics (p-value < 0.001). Only 61 PAE isolates from a total of 1827 Pseudomonas cultures (3.33%) were difficult to treat, of which only one was a bacteremia. Interestingly, we found that the carbapenem susceptibility of Pseudomonas was unaffected by the decrease in their consumption. These results augur that antimicrobial pressure may not be the sole contributor to resistance emergence. Finally, antimicrobial stewardship seems to have a positive impact on nosocomial epidemiology.

Draft Genome Sequences of Two Multidrug-Resistant Sequence Type 405 Escherichia coli Isolates without Clinical Infection.

We present the genome sequences of two carbapenemase-producing sequence type 405 Escherichia coli clinical isolates, strains Marseille-Q1950 and Marseille-Q1951. The isolates were obtained 1 month apart during the patient's hospitalization in Lebanon, in May (Marseille-Q1950) and June (Marseille-Q1951) 2019. The genome sizes of strains Marseille-Q1950 and Marseille-Q1951 were 5,181,515 bp and 5,213,451 bp, respectively.

A challenging modified measles outbreak in vaccinated healthcare providers.

BACKGROUND: Modified measles is rarely reported and thought to be an attenuated, less transmissible form of measles. The occupational safety and management of previously immunized healthcare providers (HCP) facing the global reemergence of measles is controversial and unclear.Aim: We report a measles outbreak with an unusual presentation among our vaccinated HCP at Saint George Hospital University Medical Center (SGHUMC) in Lebanon that occurred during a nationwide measles epidemic. METHODS: We recorded cases at SGHUMC, a 333-bed tertiary-care center, from April 2018 to June 2018. We established a measles clinic for investigating all febrile patients. HCP exposure was linked to influx of index cases through our Emergency Department. Modified measles was defined as any variation in the classic presentation with a pinpoint/vesicular rash, documented exposure and evidence of prior immunity. We performed serology testing to diagnose and/or document immunity and implemented outbreak controls measures including PPE, airborne isolation, and mass notification. FINDINGS: We diagnosed 8 inpatients with classic measles, and 9 affected HCP. We diagnosed 8 HCP with modified measles. One previously immunized HCP developed classic measles despite being immunized and having a positive IgG titer. Our contact tracing revealed a total of 96 exposed HCP with 27 HCP showing non-specific signs of viral illness. We required all the 9 affected HCP to undergo home isolation. CONCLUSION: We believe it is a top priority to achieve adequate measles immunity, especially among HCP that are at the frontline of healthcare systems. This necessitates revisiting vaccination schedules and achieving seroprotective titers to reclaim proper herd immunity.

Trends of Multidrug-Resistant Pathogens, Difficult to Treat Bloodstream Infections, and Antimicrobial Consumption at a Tertiary Care Center in Lebanon from 2015-2020: COVID-19 Aftermath.

INTRODUCTION: We studied the trend of antimicrobial resistance and consumption at Saint George Hospital University Medical Center (SGHUMC), a tertiary care center in Beirut, Lebanon, with a focus on the SARS-CoV-2 pandemic. MATERIALS AND METHODS: We calculated the isolation density/1000 patient-days (PD) of the most isolated organisms from 1 January 2015-31 December 2020 that included: E. coli (Eco), K. pneumoniae (Kp), P. aeruginosa (Pae), A. baumannii (Ab), S. aureus (Sau), and E. faecium (Efm). We considered March-December 2020 a surrogate of COVID-19. We considered one culture/patient for each antimicrobial susceptibility and excluded Staphylococcus epidermidis, Staphylococcus coagulase-negative, and Corynebacterium species. We analyzed the trends of the overall isolates, the antimicrobial susceptibilities of blood isolates (BSI), difficult-to-treat (DTR) BSI, carbapenem-resistant Enterobacteriaceae (CRE) BSI, and restricted antimicrobial consumption as daily-defined-dose/1000 PD. DTR implies resistance to carbapenems, beta-lactams, fluoroquinolones, and additional antimicrobials where applicable. RESULTS AND DISCUSSION: After applying exclusion criteria, we analyzed 1614 blood cultures out of 8314 cultures. We isolated 85 species, most commonly Eco, at 52%. The isolation density of total BSI in 2020 decreased by 16%: 82 patients were spared from bacteremia, with 13 being DTR. The isolation density of CRE BSI/1000 PD decreased by 64% from 2019 to 2020, while VREfm BSI decreased by 34%. There was a significant decrease of 80% in Ab isolates (p-value < 0.0001). During COVID-19, restricted antimicrobial consumption decreased to 175 DDD/1000 PD (p-value < 0.0001). Total carbapenem consumption persistently decreased by 71.2% from 108DDD/1000 PD in 2015-2019 to 31 DDD/1000 PD in 2020. At SGHUMC, existing epidemics were not worsened by the pandemic. We attribute this to our unique and dynamic collaboration of antimicrobial stewardship, infection prevention and control, and infectious disease consultation.

Draft Genome Sequence of an NDM-1-Producing Sequence Type 101 (ST101) Klebsiella pneumoniae Strain, Marseille-Q1949.

A pan-drug-resistant Klebsiella pneumoniae strain was isolated from the blood of a 70-year-old critically ill patient in April 2019. Interestingly, the patient recovered and was discharged home a month later. The genome of strain Marseille-Q1949 is 5,607,584 bp long and has a 57.1% G+C content and 5,467 protein-coding genes.

SCA Medium: A New Culture Medium for the Isolation of All Candida auris Clades.

Candida auris is an emerging multidrug-resistant yeast causing nosocomial infections and associated with high mortality in immunocompromised patients. Rapid identification and characterisation are necessary for diagnosis and containing its spread. In this study, we present a selective culture medium for all C. auris clades. This medium is sensitive with a limit of detection ranging between 101 and 102 CFU/mL. The 100% specificity of SCA (specific C. auris) medium is confirmed on a set of 135 Candida strains, 50 bacterial species and 200 human stool samples. Thus, this medium specifically selects for C. auris isolation from clinical samples, allowing the latter to study its phenotypic profile.

Implementation of an in-house real-time reverse transcription-PCR assay for the rapid detection of the SARS-CoV-2 Marseille-4 variant.

INTRODUCTION: The SARS-CoV-2 pandemic has been associated with the occurrence since summer 2020 of several viral variants that overlapped or succeeded each other in time. Those of current concern harbor mutations within the spike receptor binding domain (RBD) that may be associated with viral escape to immune responses. In our geographical area a viral variant we named Marseille-4 harbors a S477 N substitution in this RBD. MATERIALS AND METHODS: We aimed to implement an in-house one-step real-time reverse transcription-PCR (qPCR) assay with a hydrolysis probe that specifically detects the SARS-CoV-2 Marseille-4 variant. RESULTS: All 6 cDNA samples from Marseille-4 variant strains identified in our institute by genome next-generation sequencing (NGS) tested positive using our Marseille-4 specific qPCR, whereas all 32 cDNA samples from other variants tested negative. In addition, 39/42 (93 %) respiratory samples identified by NGS as containing a Marseille-4 variant strain and 0/26 samples identified as containing non-Marseille-4 variant strains were positive. Finally, 2018/3960 (51%) patients SARS-CoV-2-diagnosed in our institute, 10/277 (3.6 %) respiratory samples collected in Algeria, and none of 207 respiratory samples collected in Senegal, Morocco, or Lebanon tested positive using our Marseille-4 specific qPCR. DISCUSSION: Our in-house qPCR system was found reliable to detect specifically the Marseille-4 variant and allowed estimating it is involved in about half of our SARS-CoV-2 diagnoses since December 2020. Such approach allows the real-time surveillance of SARS-CoV-2 variants, which is warranted to monitor and assess their epidemiological and clinical characterics based on comprehensive sets of data.

Carbapenem resistant organisms: A 9-year surveillance and trends at Saint George University Medical Center.

INTRODUCTION: Carbapenem resistant organisms (CRO) constitute a large group of bacteria with different mechanisms of resistance and recently increasing global incidence. This rise has ambiguous dynamics and essential local epidemiologic data is lacking. MATERIALS AND METHODS: In this retrospective study at the 400-bed Saint George Hospital (SGH) in Beirut, Lebanon, we retrieved electronic laboratory records of all intrinsic and acquired CRO isolates from January 1, 2010 until June 30, 2018. Isolation density was calculated as: number of isolates/1000PD. Analysis carried out using WHOnet with a trend time series analysis. RESULTS: During the study period, a total of 2150 non-duplicate CRO were isolated. While Acinetobacter baumanii (AB), Pseudomonas aeruginosa (CRPa), and Stenotrophomonas maltophilia (Sm) constituted 85% of total CRO in the study period, the carbapenem resistant enterobacteriaceae (CRE) rose from few sporadic cases before 2016 to a solid 32% of total CRO in 2018. Our most concrete trends were as follows. The rate of AB bacteremia was at an average of 0.114/1000 PD from 2011 to 2014. In 2015, a sudden doubling of AB bacteremia to 0.23/1000 PD. In 2017, there was a significant decrease to 0.113/1,000PD (p < 0.0001) to reach 0.097/1000PD in 2018 with a continuously declining trend. The peak of Sm bacteremia was in 2016 at 0.121/1000PD after which it significantly decreased by 21% in 2017 to disappear in 2018 (p < 0.0001). There were no significant trends observed in the isolation density of the CRPa group from 2010 until June 2018. Klebsiella pneumonia (CRKp) bacteremia was isolated first in 2013, then in 2016 and continued to rise (p = 0.028). In 2017, carbapenem resistant KP bacteremia rate doubled to 0.05/1,000PD from 0.024/1000 (p = 0.0139). CONCLUSION: In conclusion, this 9-year study at SGH depicts the major trends and dynamics of local CRO isolation, mainly A. baumanii, P. aeruginosa and CRKp. Further efforts are warranted both locally and internationally for a richer understanding of this trend. Bearing in mind that understanding antimicrobial resistance is a complex, multifaceted process that is only feasible when all its aspects are combined: molecular, phenotypic and clinical.

Control and Elimination of Extensively Drug-Resistant Acinetobacter baumanii in an Intensive Care Unit.

We decreased antimicrobial drug consumption in an intensive care unit in Lebanon by changing to colistin monotherapy for extensively drug-resistant Acinetobacter baumanii infections. We saw a 78% decrease of A. baumanii in sputum and near-elimination of blaoxa-23-carrying sequence type 2 clone over the 1-year study. Non-A. baumanii multidrug-resistant infections remained stable.

First Detection of Colistin-Resistant Klebsiella pneumoniae in Association with NDM-5 Carbapenemase Isolated from Clinical Lebanese Patients.

This study aims to describe the molecular mechanisms of carbapenem and colistin resistance in Klebsiella pneumoniae strains isolated from hospitalized patients in Lebanon. We report in this study the first description of NDM-5 producing carbapenem-resistant K. pneumoniae ST383, as well as the presence of two out of five isolates resistant to colistin due to mutations in the amino acid sequences of proteins (PmrB, PhoQ, and MgrB). Therefore, screening of such isolates may be effective in limiting the spread of these resistant microorganisms in hospitalized patients and within the community.