Nationwide proficiency assessment of bacterial identification and antimicrobial susceptibility testing among 110 laboratories in Lebanon.

INTRODUCTION: Proficiency testing (PT) is one of the most valuable and important activities for the Clinical Microbiology Laboratories (CML) to enroll in to ensure the accuracy and reliability of results. This first time conducted nationwide study was warranted to assess the PT performance activity among CML in Lebanon. METHODOLOGY: Four training and PT activities were organized for 110 nationwide laboratories involved in providing clinical microbiology services. In each PT activity, five different bacterial species were distributed to each laboratory to provide identification (ID) and antimicrobial susceptibility testing (AMST) according to prior discussions and guidelines. RESULTS: The percentages of labs that correctly identified the bacterial species and performed the relevant AMST to it, respectively, were as follows: S. aureus, (100% and 67.8%); Enterococcus faecalis (71% and 82%); Listeria monocytogenes (75% and 61%); Streptococcus agalactiae (86% and 71%); Corynebacterium amycolatum (7% and 33 %); Pseudomonas aeruginosa, (93 % and 53.4%); Klebsiella pneumoniae, (97% and 67.7%); Salmonella typhi ESBL (87 % and 66%); Enterobacter aerogenes (89% and 59%) and Stenotrophomonas maltophilia (84 % and 65%). The resistant types for the species were specified by labs as carbapenem resistant (CR) K. pneumoniae in 78 %, CR E. aerogenes in 34 %, MRSA in 83 %, and VRE in 80.5%. CONCLUSIONS: The wide variation as well as the overall humble scoring of accurate results reflects the dire need for the MOPH to establish and maintain a PT activity program, and entrust the reference laboratory to provide continuing education and training sessions.

Impact of Different Irrigant Agitation Methods on Bacterial Elimination from Infected Root Canals.

Activation techniques are essential for root canal disinfection but may result in incomplete removal of bacteria. The aim of our study was to assess the antibacterial action of sonically, ultrasonically and laser-activated irrigation and 5.25% sodium hypochlorite (NaOCl) on Enterococcus faecalis in an infected tooth. Forty-four extracted mandibular premolars were mechanically prepared, sterilized, and inoculated with E. faecalis for 1 week. Bacterial counts after inoculation were evaluated in 4 randomly chosen teeth, remaining root canals were divided into 4 groups. Group A: laser-activated irrigation by photon-induced photoacoustic streaming, Group B: the sonic irrigation by EDDY, Group C: ultrasonic irrigation by EndoUltra, and Group D: 5.25% NaOCl. Colony forming unit (CFU) counts were measured and Kolmogorov-Smirnov, Wilcoxon, Kruskal-Wallis and Mann-Whitney tests were used to determine differences. The mean of CFU was found to significantly decrease in group D, 2110 ± 1015.93 (p < 0.001). Changes in measurement levels followed the same trend over time in groups A 27.40 ± 30.15, B 81.3 ± 85.68 and C 44.40 ± 67.12 (p = 0.141). The average CFU after irrigation in all groups was significantly greater than 0. Within the limitations of this study, all activation techniques were superior to NaOCl 5.25% in reducing E. faecalis from the infected tooth model.

Post Syrian war impact on susceptibility rates and trends in molecular characterization of Enterobacteriaceae.

OBJECTIVE: Describe susceptibility and molecular profiles among Enterobacteriaceae pathogens and to explore if war, among other factors, can affect antimicrobial resistance. METHODS: Clinical isolates from the Study for Monitoring Antimicrobial Resistance Trends associated with urinary tract and intra-abdominal infections between 2011 and 2015 were identified in Lebanon and Jordan. Susceptibility testing and molecular characterization were performed as per standard methods. RESULTS: A total of 1486 Enterobacteriaceae pathogens (including unusual pathogens) were identified. Incidence rates of  extended spectrum ß-lactamases were high with an overall higher prevalence of resistance in Jordan compared with Lebanon. CTX-M-15 was the most prevalent extended spectrum ß-lactamases produced and OXA-48 the most reported carbapenemases subtype. CONCLUSION: Changes in healthcare system due to war could impact regional resistance patterns and which requires a continuous surveillance program and containment plan.

Clonal emergence of Klebsiella pneumoniae ST14 co-producing OXA-48-type and NDM carbapenemases with high rate of colistin resistance in Dubai, United Arab Emirates.

Few studies have addressed the molecular epidemiology of carbapenem-resistant Enterobacteriaceae (CRE) isolates in the Arabian Peninsula, and such investigations have been missing from Dubai, a major economical, tourism and medical centre of the region. The antibiotic susceptibility, the carbapenemase type produced, and the clonality of 89 CRE strains isolated in five major Dubai hospitals in June 2015 to June 2016 were determined. Thirty-three percent of the collection of 70 Klebsiella pneumoniae, 13 Escherichia coli and 6 other Enterobacteriaceae were extremely drug resistant, 27% were resistant to colistin, and 4.5% (4 K. pneumoniae isolates) were resistant to all antibiotics tested. The colistin resistance rate in K. pneumoniae was 31.4%. None of the isolates carried mobile colistin resistance genes. Seventy-seven isolates produced carbapenemase: 53.3% OXA-48-like, 24.7% NDM and 22.1% both OXA-48-like and NDM, respectively. Pulsed-field gel electrophoresis clustered 50% of K. pneumoniae into a 35-membered group, which showed significant association with double carbapenemase production, with extreme drug resistance, and with being isolated from Emirati patients. Members of the cluster belonged to sequence type ST14. The rate of colistin resistance in K. pneumoniae ST14 was 37.1% vs. 27.1% of K. pneumoniae isolates outside of the cluster. Two of the panresistant K. pneumoniae isolates also belonged to ST14, whereas the other two were ST15 and ST231, respectively. In conclusion, beyond the overall high colistin resistance rate in CRE, the emergence of a highly resistant clone of K. pneumoniae ST14 in all Dubai hospitals investigated is a serious problem requiring immediate attention.

In vitro Evaluation of the Colistin-Carbapenem Combination in Clinical Isolates of A. baumannii Using the Checkerboard, Etest, and Time-Kill Curve Techniques.

The worldwide increase in the emergence of carbapenem resistant Acinetobacter baumannii (CRAB) calls for the investigation into alternative approaches for treatment. This study aims to evaluate colistin-carbapenem combinations against Acinetobacter spp., in order to potentially reduce the need for high concentrations of antibiotics in therapy. This study was conducted on 100 non-duplicate Acinetobacter isolates that were collected from different patients admitted at Saint George Hospital-University Medical Center in Beirut. The isolates were identified using API 20NE strips, which contain the necessary agents to cover a panel of biochemical tests, and confirmed by PCR amplification of blaOXA-51-like. Activities of colistin, meropenem and imipenem against Acinetobacter isolates were determined by ETEST and microdilution methods, and interpreted according to the guidelines of the Clinical and Laboratory Standards Institute. In addition, PCR amplifications of the most common beta lactamases contributing to carbapenem resistance were performed. Tri locus PCR-typing was also performed to determine the international clonality of the isolates. Checkerboard, ETEST and time kill curves were then performed to determine the effect of the colistin-carbapenem combinations. The synergistic potential of the combination was then determined by calculating the Fractional Inhibitory Concentration Index (FICI), which is an index that indicates additivity, synergism, or antagonism between the antimicrobial agents. In this study, 84% of the isolates were resistant to meropenem, 78% to imipenem, and only one strain was resistant to colistin. 79% of the isolates harbored blaOXA-23-like and pertained to the International Clone II. An additive effect for the colistin-carbapenem combination was observed using all three methods. The combination of colistin-meropenem showed better effects as compared to colistin-imipenem (p < 0.05). The colistin-meropenem and colistin-imipenem combinations also showed a decrease of 2.6 and 2.8-fold, respectively in the MIC of colistin (p < 0.001). Time kill assays additionally showed synergistic effects for a few isolates, and no bacterial re-growth was detected following a 24 h incubation. Our study showed that the combination of colistin with carbapenems could be a promising antimicrobial strategy in treating CRAB infections and potentially lowering colistin toxicity related to higher doses used in colistin monotherapy.