Importance of consultations using mobile teams in the screening and treatment of neglected tropical skin diseases in Benin.

CONTEXT: Since 2013, the World Health Organization has recommended integrated control strategies for neglected tropical diseases (NTDs) with skin manifestations. We evaluated the implementation of an integrated approach to the early detection and rapid treatment of skin NTDs based on mobile clinics in the Ouémé and Plateau areas of Benin. METHODS: This descriptive cross-sectional study was performed in Ouémé and Plateau in Benin from 2018 to 2020. Consultations using mobile teams were performed at various sites selected by reasoned choice based on the epidemiological data of the National Program for the Control of Leprosy and Buruli Ulcer. All individuals presenting with a dermatological lesion who voluntarily approached the multidisciplinary management team on the day of consultation were included. The information collected was kept strictly anonymous and was entered into an Excel 2013 spreadsheet and analyzed with Stata 11 software. RESULTS: In total, 5,267 patients with various skin conditions consulted the medical team. The median age of these patients was 14 years (IQR: 7-34 years). We saw 646 (12.3%) patients presenting NTDs with skin manifestations, principally scabies, in 88.4% (571/646), followed by 37 cases of Buruli ulcer (5.8%), 22 cases of leprosy (3.4%), 15 cases of lymphatic filariasis (2.3%) and one case of mycetoma (0.2%). We detected no cases of yaws. CONCLUSION: This sustainable approach could help to decrease the burden of skin NTDs in resource-limited countries.

An Overview of 10 Years of Activity of a Molecular Laboratory for Buruli Ulcer Diagnosis at a Field Hospital in Benin.

Buruli ulcer (BU) is a neglected tropical disease caused by Mycobacterium ulcerans. Early diagnosis is crucial to prevent morbidity. In November 2012, a field laboratory fully equipped for the rapid on-site quantitative PCR (qPCR) diagnosis of M. ulcerans was established at the Buruli ulcer treatment center (CDTLUB) center in Pobè Benin, a region where BU is endemic. We describe its first 10 years of activity and its gradual evolution into an expert laboratory for BU diagnosis. From 2012 to 2022, the laboratory analyzed 3,018 samples from patients attending consultations for suspected BU at the CDTLUB in Pobè. Ziehl-Neelsen staining and qPCR targeting the IS2404 sequence were performed. Since 2019, the laboratory has also received and analyzed 570 samples from other centers. The laboratory confirmed the diagnosis of BU by qPCR for 39.7% samples: M. ulcerans DNA was detected in 34.7% of swabs, 47.2% of all fine needle aspiration samples (FNA) and 44.6% of all skin biopsy specimens. Positive Ziehl-Neelsen staining results were obtained for 19.0% samples. Bacterial load, estimated by qPCR, was significantly greater for the Ziehl-Neelsen-positive samples than for Ziehl-Neelsen-negative samples, and detection rates were highest for FNA samples. Overall, 26.3% of the samples received from other centers were positive for BU. Most of these samples were sent by the CDTLUBs of Lalo, Allada, and Zagnanado, Benin. The establishment of the laboratory in the CDTLUB of Pobè has been a huge success. Optimal patient care depends on the close proximity of a molecular biology structure to BU treatment centers. Finally, FNA should be promoted among caregivers. IMPORTANCE Here, we describe the first 10 years of activity at a field laboratory established at the Buruli ulcer treatment center (CDTLUB) in Pobè, Benin, a country in which Mycobacterium ulcerans is endemic. Between 2012 and 2022, the laboratory analyzed 3,018 samples from patients consulting the CDTLUB of Pobè with a suspected clinical BU. Ziehl-Neelsen staining and qPCR targeting the IS2404 sequence were performed. In total, 39.7% of samples tested positive by qPCR and 19.0% tested positive by Ziehl-Neelsen staining. Detection rates were highest for FNA samples, and the bacterial loads estimated by qPCR were significantly higher for Ziehl-Neelsen-positive samples than for Ziehl-Neelsen-negative samples. Since 2019, the laboratory has also analyzed 570 samples received from outside the CDTLUB of Pobè, 26.3% of which were positive for BU. Most of these samples were sent by the CDTLUBs of Lalo, Allada, and Zagnanado in Benin. The establishment of the laboratory in the CDTLUB of Pobè has been a huge success, with major benefits for both the medical staff and patients. Our findings illustrate that the usefulness and feasibility of having a diagnostic center in rural Africa, where the disease is endemic, is a key part of optimal patient care, and that FNA should be promoted to increase detection rates.

Rejuvenating the Activity of Usual Antibiotics on Resistant Gram-Negative Bacteria: Recent Issues and Perspectives.

Antibiotic resistance continues to evolve and spread beyond all boundaries, resulting in an increase in morbidity and mortality for non-curable infectious diseases. Due to the failure of conventional antimicrobial therapy and the lack of introduction of a novel class of antibiotics, novel strategies have recently emerged to combat these multidrug-resistant infectious microorganisms. In this review, we highlight the development of effective antibiotic combinations and of antibiotics with non-antibiotic activity-enhancing compounds to address the widespread emergence of antibiotic-resistant strains.

clbP Gene, a Potential New Member of the ß-Lactamase Family.

The colibactin island (pks) of Escherichia coli formed by 19 genes (55-Kb), encodes non-ribosomal peptide (NRP) and polyketide (PK) synthases, which allow the synthesis of colibactin, a suspected hybrid PK-NRP compound that causes damage to DNA in eukaryotic cells. The clbP, an unusual essential gene, is found in the operon structure with the clbS gene in the pks-encoded machinery. Interestingly, the clbP gene has been annotated as a ß-lactamase but no previous study has reported its ß-lactamase characteristics. In this study, we (i) investigated the ß-lactamase properties of the clbP gene in silico by analysing its phylogenetic relationship with bacterial ß-lactamase and peptidase enzymes, (ii) compared its three-dimensional (3D) protein structure with those of bacterial ß-lactamase proteins using the Phyr2 database and PyMOL software, and (iii) evaluated in vitro its putative enzymatic activities, including ß-lactamase, nuclease, and ribonuclease using protein expression and purification from an E. coli BL21 strain. In this study, we reveal a structural configuration of toxin/antitoxin systems in this island. Thus, similar to the toxin/antitoxin systems, the role of the clbP gene within the pks-island gene group appears as an antitoxin, insofar as it is responsible for the activation of the toxin, which is colibactin. In silico, our analyses revealed that ClbP belonged to the superfamily of ß-lactamase, class C. Furthermore, in vitro we were unable to demonstrate its ß-lactamase activity, likely due to the fact that the clbP gene requires co-expression with other genes, such as the genes present in the pks-island (19 genes). More research is needed to better understand its actions, particularly with regards to antibiotics, and to discover whether it has any additional functions due to the importance of this gene and its toxicity.

Clonal Dissemination of Plasmid-Mediated Carbapenem and Colistin Resistance in Refugees Living in Overcrowded Camps in North Lebanon.

Carbapenem and colistin-resistant bacteria represent a global public health problem. Refugees carrying these bacteria and living in inadequate shelters can spread these microorganisms. The aim of this study was to investigate the intestinal carriage of these bacteria in Syrian refugees in Lebanon. Between June and July 2019, 250 rectal swabs were collected from two refugee camps in North Lebanon. Swabs were cultured on different selective media. Antibiotic susceptibility testing was performed using the disk diffusion method. Carbapenemase-encoding genes and mcr genes were investigated using real-time polymerase chain reaction (RT-PCR) and standard polymerase chain reaction (PCR). Epidemiological relatedness was studied using multilocus sequence typing (MLST). From 250 rectal swabs, 16 carbapenem-resistant, 5 colistin-resistant, and 4 colistin and carbapenem-resistant Enterobacteriaceae were isolated. The isolates exhibited multidrug-resistant phenotypes. Seven Klebsiella pneumoniae isolates harboured the blaOXA-48 gene, and in addition four K. pneumoniae had mutations in the two component systems pmrA/pmrB, phoP/phoQ and co-harboured the blaNDM-1 gene. Moreover, the blaNDM-1 gene was detected in six Escherichia coli and three Enterobacter cloacae isolates. The remaining five E. coli isolates harboured the mcr-1 gene. MLST results showed several sequence types, with a remarkable clonal dissemination. An urgent strategy needs to be adopted in order to avoid the spread of such resistance in highly crowded underserved communities.

Carbapenem and colistin-resistant bacteria in North Lebanon: Coexistence of mcr-1 and NDM-4 genes in Escherichia coli.

INTRODUCTION: The increasing incidence of infections caused by multidrug-resistant bacteria is considered a global health problem. This study aimed to investigate this resistance in Gram-negative bacteria isolated from patients hospitalized in North-Lebanon. METHODOLOGY: All isolates were identified using the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Antibiotic susceptibility testing was achieved using disk diffusion, E-test and Broth microdilution methods. Phenotypic detection of carbapenemase was carried out using the CarbaNP test. RT-PCR, standard-PCR and sequencing were performed to detect resistance genes and oprD gene. Conjugal transfer was carried out between our isolates and Escherichia coli J53 to detect the genetic localization of resistance genes. MLST was conducted to determine the genotype of each isolate. RESULTS: Twenty-three carbapenem-resistant Enterobacterales of which eight colistin-resistant Escherichia coli, and Twenty carbapenem-resistant Pseudomonas aeruginosa were isolated. All isolates showed an imipenem MIC greater than 32 mg/mL with MICs for colistin greater than 2 mg/L for E. coli isolates. All the Enterobacterales isolates had at least one carbapenemase-encoding gene, with E. coli isolates coharboring blaNDM-4 and mcr-1 genes. Moreover, 16/20 Pseudomonas aeruginosa harbored the blaVIM-2 gene and 18/20 had mutations in the oprD gene. MLST revealed that the isolates belonged to several clones. CONCLUSIONS: We report here the first description in the world of clinical E. coli isolates coharboring blaNDM-4 and mcr-1 genes, and K. pneumoniae isolates producing NDM-6 and OXA-48 carbapenemases. Also, we describe the emergence of NDM-1-producing E. cloacae in Lebanon. Screening for these isolates is necessary to limit the spread of resistant microorganisms in hospitals.

Bacterial infection during wars, conflicts and post-natural disasters in Asia and the Middle East: a narrative review.

Introduction: Bacterial infections resulting from wars and natural disasters represent a major public health problem. Over the past 50 years, Asia and the Middle East have suffered several wars. Moreover, East-Asian countries are considered the most natural disaster-prone countries in the world.Areas covered: This review focuses on bacterial infection occurring during wars and after natural disasters, among refugees, wounded citizens and soldiers as well as the prevention and control measures that must be taken.Expert opinion: During wars, refugees and soldiers represent the two main sources of bacterial infections. Refugees coming from countries with a high prevalence of antimicrobial resistance can spread these pathogens to their final destination. In addition, these refugees living in inadequate shelters can contribute to the spread of bacterial infections. Moreover, some factors including the presence of fixed imported fragments; environmental contamination and nosocomial transmissions, play a key role in the dissemination of bacteria among soldiers. As for natural disasters, several factors are associated with increased bacterial transmissions such as the displacement of large numbers of people into over-crowded shelters, high exposure to disease vectors, lack of water and sanitation. Here, we carry out a systematic review of the bacterial infections that follow these two phenomena.

Intestinal carriage of colistin-resistant Enterobacteriaceae at Saint Georges Hospital in Lebanon.

OBJECTIVES: The increase in resistance to antibiotics has led to the revival of colistin as the last option for treatment, which automatically led to an increase of colistin-resistant, Gram-negative bacteria. In this study, we report the presence of clinical colistin-resistant Enterobacteriaceae isolated from a Lebanese hospital. METHODS: From 23 rectal swabs, eight colistin-resistant clinical strains (five Escherichia coli, two Enterobacter cloacae, and one Klebsiella pneumoniae) were isolated. Antibiotic susceptibility testing was performed using the disk diffusion method and Etest. The broth microdilution method was used to determine colistin susceptibility. Reverse transcription polymerase chain reaction (RT-PCR), standard PCR and sequencing were used to investigate genes encoding for extended-spectrum ß-lactamases, carbapenemases and colistin resistance. Genotyping of these isolates was conducted by multilocus sequence typing (MLST). RESULTS: Results of antibiotic susceptibility testing revealed that all isolates were resistant to colistin. They had MICs for colistin that ranged from 8 to 32 mg/L. Real-time PCR results showed that five strains harboured blaTEM-1 and one strain harboured blaTEM-163. Moreover, four strains were positive for blaCTX-M-15, blaCTX-M-103 and blaCTX-M-189, and K. pneumoniae harboured blaSHV-1. Observed colistin resistance was linked to amino acid substitutions into protein sequences of pmrA/B, phoP/Q, and mgrB. Interestingly, we report here a mutation in the mgrB regulator and pmrA/B, phoP/Q in colistin-resistant E. cloacae and E. coli clinical isolates for the first time in Lebanon. CONCLUSION: This study highlights the presence of colistin-resistant Gram-negative bacteria in a Lebanese hospital, which is worrisome. An urgent strategy needs to be adopted to avoid the spread of such bacteria.

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.

Investigation of multidrug-resistant ST2 Acinetobacter baumannii isolated from Saint George hospital in Lebanon.

BACKGROUND: Acinetobacter baumannii is an opportunistic pathogen causing various nosocomial infections. The spread of multidrug-resistant A. baumannii is a major public health problem. The aim of this study was to investigate the molecular epidemiology and the genetic support of multidrug-resistant A. baumannii isolates collected from Saint-Georges Hospital in Lebanon. METHODS: Between January and August 2016, 31 A. baumannii isolates were collected from sputum samples of patients infected with ventilator-associated pneumonia (VAP) and treated with colistin-carbapenem combination therapy. Antibiotic susceptibility testing was performed using the disk diffusion method. Carbapenemases, extended spectrum ß-lactamases encoding genes and mcr-1/2 genes were investigated by RT-PCR and standard PCR. The epidemiological relatedness of the strains was studied using MLST analysis. RESULTS: Most of the isolates exhibited multidrug-resistant phenotypes. All the isolates were carbapenem-resistant and among them, 30 carried the class D carbapenemase blaoxa-23 gene while one isolate carried blaoxa-72 gene. MLST results revealed three sequence types, namely ST2, ST699, and ST627. Isolates having ST2 were the most prevalent clone (29/31, 93.5%). CONCLUSIONS: This study shows a nosocomial spread of multidrug-resistant A. baumannii ST2 having blaOXA-23 gene in Saint-George in Lebanon. Monitoring and control measures need to be adopted to avoid the spread of A. baumannii to patients.

First Detection of VIM-4-Producing Pseudomonas aeruginosa and OXA-48-Producing Klebsiella pneumoniae in Northeastern (Annaba, Skikda) Algeria.

PURPOSE: The aim of the study was to investigate the prevalence and molecular support of carbapenem resistance in gram-negative bacilli clinical isolates collected between March 2013 and March 2015 in three cities (Annaba, Skikda, and Guelma) in northeastern Algeria. RESULTS: One hundred eighty-six isolates were identified as Enterobacteriaceae (161), Pseudomonas aeruginosa (18), and Acinetobacter baumannii (7). Thirty-six of 186 (19.3%) were resistant to carbapenems. Among them, 11 harbored carbapenemase genes, including blaOXA-48 (2 Klebsiella pneumoniae), blaVIM-4 (2 P. aeruginosa), blaNDM-1 (2 A. baumannii), and blaOXA-23 (5 A. baumannii). In addition, other ß-lactamases were detected: blaCTX-M-(15/66/139), blaSHV-(28/85/1/133), and blaTEM-1. All imipenem-resistant P. aeruginosa displayed OprD mutations. Multilocus sequence typing demonstrated the presence of ST 404 and ST 219 in K. pneumoniae, ST 2 and ST 85 in A. baumannii, and ST (244, 1076, 241, 227, and 233) in P. aeruginosa. CONCLUSION: In this study, we report the first detection of P. aeruginosa ST 1076 harboring the blaVIM-4 gene in African countries in two cities (Annaba and Skikda) in northeastern Algeria. Additionally, we report the first detection of blaOXA-48 in K. pneumoniae ST 404 and ST 219 in Algerian cities (Annaba and Skikda).

Prevalence and emergence of carbapenemases-producing Gram-negative bacteria in Mediterranean basin.

The emergence and the global spread of carbapenemases concern to health services worldwide. Their celestial rise among Gram-negative bacilli has challenged both the scientific and pharmaceutical sectors. Indeed, infections caused by these bacteria have limited treatment options and have been associated with high mortality and morbidity rates. Carbapenemase producers are mainly identified among Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, and Acinetobacter baumannii and still mostly in hospital settings and rarely in the community. They are closely related to KPC, VIM, IMP, NDM, and OXA-48 types. The encoding genes are mostly plasmid located and associated with various mobile genetic elements. The Mediterranean area is of interest due to a great diversity and population mixing. The prevalence of carbapenemases is particularly high and variant among countries, partially depending on the population exchange relationship between the regions and the possible reservoirs of each carbapenemase. This review described the epidemiology of carbapenemases in this region of the world highlighting the worrisome situation and the need to screen and detect these enzymes to prevent and control their dissemination especially as it is clear that very few novel antibiotics will be introduced in the next few years, making the dissemination of carbapenem-resistant Gram-negative bacteria of crucial importance worldwide.

Molecular epidemiology of environmental and clinical carbapenemase-producing Gram-negative bacilli from hospitals in Guelma, Algeria: Multiple genetic lineages and first report of OXA-48 in Enterobacter cloacae.

This study was designed to investigate environmental colonisation in Algerian hospitals by carbapenem-resistant Gram-negative bacilli (GNB), including molecular characterisation of their resistance, and to perform a comparative molecular analysis between clinical and environmental strains. GNB isolated from hospitalised patients and the hospital environment were identified using microbiological methods and matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF/MS). Antibiotic susceptibility testing was performed by disk diffusion and Etest methods. Carbapenemase- and extended-spectrum ß-lactamase (ESBL)-encoding genes were searched for using PCR and sequencing. Clonality of the environmental and clinical strains was assessed by multilocus sequencing typing (MLST). A total of 32 carbapenem-resistant GNB were isolated, including 16 (29%) of 56 multidrug-resistant (MDR) GNB from clinical specimens and 16 (48%) of 33 MDR-GNB from inanimate surfaces. Of the 32 carbapenem-resistant isolates, 14 produced a carbapenemase. The blaOXA-48 gene was detected both in clinical and surface isolates of Klebsiella pneumoniae (n=3) and Enterobacter cloacae (n=2). Clinical and surface isolates of Acinetobacter baumannii were found to produce the carbapenemases NDM-1 (7 isolates) and OXA-23 (2 isolates). MLST revealed clonal diversity and a relationship between environmental and clinical strains with identical sequence types. Here we report the first description of an OXA-48-producing E. cloacae isolate in Algeria. We also highlight the important role of inanimate surfaces in the spread of carbapenem-resistant bacteria and the emergence of nosocomial infections.

Carbapenemases and extended-spectrum ß-lactamases producing Enterobacteriaceae isolated from Tunisian and Libyan hospitals.

INTRODUCTION: The aim of the study was to investigate the prevalence of extended-spectrum ß-lactamase (ESBL) and carbapenemase production among clinical isolates of Enterobacteriaceae recovered from Tunisian and Libyan hospitals. METHODOLOGY: Bacterial isolates were recovered from patients in intensive care units and identified by biochemical tests and MALDI-TOF. Antibiotic susceptibility testing was performed by disk diffusion and the E-test method. ESBL and carbapenemase activities were detected using standard microbiological tests. Antibiotic resistance-encoding genes were screened by PCR and sequencing. Clonal relationships between Klebsiella pneumoniae strains were carried out using multi-locus sequence typing (MLST). RESULTS: A total of 87 isolates were characterized, with 51 and 36, respectively, identified as E. coli and K. pneumoniae. Overall the resistance prevalence was high for aminoglycosides (> 60%), fluoroquinolones (> 80%), and extended-spectrum cephalosporins (> 94%), and was low for imipenem (11.4%). Among this collection, 58 strains (66.6%) were ESBL producers and 10 K. pneumoniae strains (11.4%) were carbapenemase producers. The antibiotic resistance-encoding genes detected were blaCTX-M-15 (51.7%), blaTEM-1 (35.6%), several variants of blaSHV (21.8%), and blaOXA-48 (11.4%). The MLST typing of K. pneumoniae isolates revealed the presence of multiple clones and three novel sequence types. Also, close relationships between the OXA-48-producing strains from Tunisia and Libya were demonstrated. CONCLUSIONS: This study is the first paper describing the emergence of carbapenemase- and ESBL-producing Enterobacteriaceae, sensitive to colistin, isolated in Tunisia and Libya. Active surveillance and testing for susceptibility to colistin should be implementing because resistance to colistin, mainly in Klebsiella, has been recently reported worldwide.

Molecular characterisation of extended-spectrum ß-lactamase- and plasmid AmpC-producing Escherichia coli strains isolated from broilers in Béjaïa, Algeria.

This study aimed to characterise the molecular support of antibiotic resistance in expanded-spectrum cephalosporin (ESC)-resistant Escherichia coli isolates recovered from healthy broilers in Béjaïa, northeast Algeria. A total of 61 intestinal swabs from slaughtered broilers from four regions in Béjaïa locality, Algeria, were collected between February and April 2014, from which 20 ESC-resistant E. coli strains were isolated. Escherichia coli isolates were identified by classical biochemical and MALDI-TOF methods. Antibiotic susceptibility testing was performed using disk diffusion and Etest methods. Screening for ß-lactamases, aminoglycoside-modifying enzyme (AME)-encoding genes and qnr determinants was performed by PCR and sequencing. Clonal relatedness was determined using molecular typing by multilocus sequence typing (MLST). Antibiotic susceptibility testing revealed that the isolates showed high rates of resistance (>90%) to amoxicillin, amoxicillin/clavulanic acid, piperacillin/tazobactam, aztreonam, ceftazidime, streptomycin, tobramycin, nalidixic acid and ciprofloxacin. Low rates of resistance were observed for kanamycin (35%), amikacin (30%), cefoxitin (20%) and cefotaxime (15%). Molecular characterisation revealed that all of the isolates expressed the blaTEM-1 gene. Fourteen of them harboured the blaSHV-12 gene, two harboured the blaCTX-M-1 gene and four isolates harboured blaCMY-2. Screening for AME-encoding genes demonstrated that all isolates contained the aadA gene. In addition, qnrA was detected as the quinolone resistance determinant in 13 isolates. MLST revealed four known sequence types (STs), including ST744, ST38, ST1011 and ST2179, as well as one new sequence type (ST5086). Here we report the first study describing the clonal diversity of extended-spectrum ß-lactamase (ESBL)- and plasmid AmpC-producing E. coli isolated from healthy broilers in Algeria.

Early detection of metallo-ß-lactamase NDM-1- and OXA-23 carbapenemase-producing Acinetobacter baumannii in Libyan hospitals.

Acinetobacter baumannii is an opportunistic pathogen causing various nosocomial infections. The aim of this study was to characterise the molecular support of carbapenem-resistant A. baumannii clinical isolates recovered from two Libyan hospitals. Bacterial isolates were identified by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF/MS). Antibiotic susceptibility testing was performed using disk diffusion and Etest methods, and carbapenem resistance determinants were studied by PCR amplification and sequencing. Multilocus sequence typing (MLST) was performed for typing of the isolates. All 36 imipenem-resistant isolates tested were identified as A. baumannii. The blaOXA-23 gene was detected in 29 strains (80.6%). The metallo-ß-lactamase blaNDM-1 gene was detected in eight isolates (22.2%), showing dissemination of multidrug-resistant (MDR) A. baumannii in Tripoli Medical Center and Burn and Plastic Surgery Hospital in Libya, including one isolate that co-expressed the blaOXA-23 gene. MLST revealed several sequence types (STs). Imipenem-resistant A. baumannii ST2 was the predominant clone (16/36; 44.4%). This study shows that NDM-1 and OXA-23 contribute to antibiotic resistance in Libyan hospitals and represents the first incidence of the association of these two carbapenemases in an autochthonous MDR A. baumannii isolated from patients in Libya, indicating that there is a longstanding infection control problem in these hospitals.

Whole-Genome Sequence of a blaOXA-48-Harboring Raoultella ornithinolytica Clinical Isolate from Lebanon.

We analyzed the whole-genome sequence of ablaOXA-48-harboringRaoultella ornithinolyticaclinical isolate from a patient in Lebanon. The size of theRaoultella ornithinolyticaCMUL058 genome was 5,622,862 bp, with a G+C content of 55.7%. We deciphered all the molecular mechanisms of antibiotic resistance, and we compared our genome to other availableR. ornithinolyticagenomes in GenBank. The resistome consisted of 9 antibiotic resistance genes, including a plasmidicblaOXA-48gene whose genetic organization is also described.

Detection of expanded-spectrum ß-lactamases in Gram-negative bacteria in the 21st century.

Emerging ß-lactamase-producing-bacteria (ESBL, AmpC and carbapenemases) have become a serious problem in our community due to their startling spread worldwide and their ability to cause infections which are difficult to treat. Diagnosis of these ß-lactamases is of clinical and epidemiological interest. Over the past 10 years, several methods have been developed aiming to rapidly detect these emerging enzymes, thus preventing their rapid spread. In this review, we describe the range of screening and detection methods (phenotypic, molecular and other) for detecting these ß-lactamases but also whole genome sequencing as a tool for detecting the genes encoding these enzymes.

Emergence of Carbapenem-Resistant Pseudomonas aeruginosa and Acinetobacter baumannii Clinical Isolates Collected from Some Libyan Hospitals.

The aim of the present study was to investigate the molecular mechanism of carbapenem resistance in Pseudomonas aeruginosa and Acinetobacter baumannii clinical isolates recovered from Libyan hospitals between April 2013 and April 2014. In total, 49 strains (24 P. aeruginosa and 25 A. baumannii) were isolated, including 21 P. aeruginosa and 22 A. baumannii isolates (87.75%) resistant to imipenem (minimum inhibitory concentrations ≥16 µg/ml). The blaVIM-2 gene was detected in 19 P. aeruginosa isolates. All imipenem-resistant P. aeruginosa isolates showed the presence of OprD mutations. Acquired OXA-carbapenemase-encoding genes were present in all A. baumannii isolates: blaOXA-23 (n=19) and blaOXA-24 (n=3). Finally, a total of 13 and 17 different sequence types were assigned to the 21 P. aeruginosa and the 22 A. baumannii carbapenem-resistant isolates, respectively. This study is the first report describing imipenem-resistant P. aeruginosa and A. baumannii isolated from patients in Libya. We report the first case of co-occurrence of blaVIM-2 with oprD porin loss in identical isolates of P. aeruginosa in Libya and demonstrate that these oprD mutations can be used as a tool to study the clonality in P. aeruginosa isolates. We also report the first identification of multidrug-resistant A. baumannii isolates harboring blaOXA-23-like, blaOXA-24-like, and blaOXA-48-like genes in Libya.