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We evaluated the in vitro activity of eravacycline and cefoperazone/sulbactam against 42 XDR and 58 PDR Acinetobacter baumannii isolates from blood and bronchoalveolar infections. The minimum and maximum MICs for eravacycline were 0.125 and 4 mg/L, respectively. The MIC50 was 2 mg/L and the MIC90 was 3 mg/L. The minimum and maximum MICs for cefoperazone/sulbactam were 24 and >256 mg/L, respectively. The MIC50 and MIC90 were both >256 mg/L. These novel agents were not adequate for the treatment of A. baumannii infections in our hospital and we recommend that mi- crobiology laboratories perform their own evaluations before including them in clinical practice.

The global vaccination against SARS-CoV-2 has highlighted the need of assessing vaccines' immunogenicity against COVID-19. To evaluate humoral immunity induced by the BNT162b2 vaccine, we enrolled health care workers at AHEPA University Hospital of Thessaloniki, Greece to measure Anti-S SARS-CoV-2, anti-RBD SARS-CoV-2 and neutralizing antibodies. A total of 955 individuals with a median age of 50 years, were included in the study. Median values of antibodies were 1947.27 BAU/mL (Abbott SARS-CoV-2 IgG II Quant), 2064.98 BAU/mL (MAGLUMI SARS-CoV-2 S-RBD IgG) and 2464.63 IU/mL (MAGLUMI SARS-CoV-2 Neutralizing Antibodies). Individuals previously infected had greater antibody responses than infection naive ones and a 7-fold higher neutralizing antibodies titre. Antibodies degreased by age but not sex. Spearman's correlation coefficient among the three assays ranged from 0.903 to 0.969. The BNT162b2 vaccine was highly immunogenic in our cohort. Further research is needed to evaluate the vaccine's immunogenicity through time as well as in different populations., Competing Interests: The authors declare no conflict of interest., (© 2022 The Author(s).)

In collaboration with the American College of Veterinary Pathologists.

Carbapenemase-producing Klebsiella pneumoniae (CPKP) emerged in Greece in 2002 and became endemic thereafter. Driven by a notable variability in the phenotypic testing results for carbapenemase production in K. pneumoniae isolates from the intensive care units (ICUs) of our hospital, we performed a study to assess the molecular epidemiology of CPKP isolated between 2016 and 2019 using pulse-field gel electrophoresis (PFGE) including isolates recovered from 165 single patients. We investigated the molecular relatedness among strains recovered from rectal surveillance cultures and from respective subsequent infections due to CPKP in the same individual (48/165 cases). For the optimal interpretation of our findings, we carried out a systematic review regarding the clonality of CPKP isolated from clinical samples in ICUs in Europe. In our study, we identified 128 distinguishable pulsotypes and 17 clusters that indicated extended dissemination of CPKP within the hospital ICU setting throughout the study period. Among the clinical isolates, 122 harbored KPC genes (74%), 2 harbored KPC+NDM (1.2%), 38 harbored NDM (23%), 1 harbored NDM+OXA-48 (0.6%), 1 harbored NDM+VIM (0.6%) and 1 harbored the VIM (0.6%) gene. Multiple CPKP strains in our hospital have achieved sustained transmission. The polyclonal endemicity of CPKP presents a further threat for the selection of pathogens resistant to last-resort antimicrobial agents.

Vaccines against SARS-CoV-2 are known to be less immunogenic for some individuals, whereas others present notably high levels of antibody production. We assessed the cellular response to BNT162b2 among individuals with low post-vaccination antibody levels as well as in a small group of individuals with high titers. Antibody levels were assessed by the Abbott SARS-CoV-2 IgG II Quant assay. The interferon-γ production of T-cells in response to SARS-CoV-2 antigens was determined using Qiagen's QuantiFERON SARS-CoV-2 ELISA test. Our results showed that participants with high antibody levels presented adequate cellular response in all studied cases, whereas those with low antibody levels generally showed limited to almost absent cellular response five months post vaccination.

Northern Greece was struck by an intense second COVID-19 (coronavirus disease 2019) epidemic wave during the fall of 2020. Because of the coinciding silent epidemic of multidrug-resistant organisms, the handling of COVID-19 patients became even more challenging. In the present study, the microbiological characteristics of bacteremias in confirmed cases of hospitalized COVID-19 patients were determined. Data from 1165 patients hospitalized between September and December 2020 were reviewed regarding the frequency of bloodstream infections, the epidemiology and the antibiotic susceptibility profiles of the causative bacteria. The hospital's antibiotic susceptibility data for all major nosocomial pathogens isolated from bacteremias of COVID-19 patients between September and December 2020 versus those between September and December 2019 were also compared. Overall, 122 patients developed bacteremia (10.47%). The average of time interval between hospitalization date and development of bacteremia was 13.98 days. Admission to ICU occurred in 98 out of 122 patients with an average stay time of 15.85 days and 90.81% in-hospital mortality. In total, 166 pathogens were recovered including 114 Gram-negative bacteria and 52 Gram-positive cocci. Acinetobacter baumannii was the most frequent ( n = 51) followed by Klebsiella pneumoniae ( n = 45) and Enterococcus faecium ( n = 31). Bacteremias in hospitalized COVID-19 patients were related with prolonged time of hospitalization and higher in-hospital mortality, and the isolated microorganisms represented the bacterial species that were present in our hospital before the COVID-19 pandemic. Worryingly, the antibiotic resistance rates were increased compared with the pre-pandemic era for all major opportunistic bacterial pathogens. The pandemic highlighted the need for continuous surveillance of patients with prolonged hospitalization., (© The Author(s) 2021. Published by Oxford University Press on behalf of FEMS.)

The present review aims to study and detect the global emergence of mcr genes in E. coli, K. pneumoniae and Salmonella spp., isolates from human specimens over the last six years. Nowadays the rise of multidrug-resistant superbugs has made essential the return of drugs that were previously abandoned. A clear example is colistin, which acts against multidrug - resistant gram - negative pathogens, including Enterobacterales. Colistin resistance is an unfortunate fact, with the emergence of mcr genes conferring resistance to colistin in Enterobacterales posing the most recent threat. Literature about mcr genes and their spread in E. coli, K. pneumoniae and Salmonella spp. is cited, focusing on the emergence of mcr genes in human specimens since 2015. The data were taken from the PubMed and Scopus databases. It seems that the mcr-1 gene continues to be the protagonist among the three species. E. coli is the dominant species harbouring mcr genes. Moreover, plasmid - mediated colistin resistance is also conferred upon other species that carry different genes resistant to antibiotics. There are only scarse reports on human Salmonella spp isolates harbouring mcr genes. Finally, the emergence of the mcr-9 gene in all of them is quite remarkable. CONCLUSION: Plasmid - mediated colistin resistance in Enterobacterales is a global issue and has been worsening over the years. The continuous mutations of mcr gene subtypes underline the need for better surveillance, constant investigation and wise use of colistin, especially in countries with high levels of antibiotic resistance., (Copyright © 2021 by Academy of Sciences and Arts of Bosnia and Herzegovina.)

The increased demand for SARS-CoV-2 molecular testing during the COVID-19 pandemic resulted in shortage of reagents and consumables. Pooling of specimens could be an alternative strategy to overcome these problems. Initial evaluation of the pooling strategy was performed using known positive specimens, previously tested individually, and their respective pools of plus four (5X), five (6X) and nine (10X) known negative specimens. Subsequently, 35 positive 5X and 35 positive 6X pools containing only one positive specimen per pool were analyzed prospectively regarding the difference in Ct values in pooled versus individual specimens. When the number of samples in the pool were five or six, the average deviation of Ct differences was < 1; therefore, this strategy was followed in the prospective study. Significant difference in Ct values was observed in positive specimens when tested individually and in 5X pools ( p  = 0.006), while the difference was not significant when positive specimens were tested individually and in 6X pools ( p  = 0.07). The difference in Ct values was not significant between the 5X and 6X pools. Testing in pools of five or six specimens is a reliable option for SARS-CoV-2 RNA detection when mass testing is needed., Competing Interests: Conflict of interestThe authors declare that there is no conflict of interest., (© Indian Virological Society 2021.)

Nasopharyngeal swab specimen (NPS) molecular testing is considered the gold standard for SARS-CoV-2 detection. However, saliva is an attractive, noninvasive specimen alternative. The aim of the study was to evaluate the diagnostic accuracy of Advanta Dx SARS-CoV-2 RT-PCR saliva-based assay against paired NPS tested with either NeumoDx TM SARS-CoV-2 assay or Abbott Real Time SARS-CoV-2 assay as the reference method. We prospectively evaluated the method in two settings: a diagnostic outpatient and a healthcare worker screening convenience sample, collected in November-December 2020. SARS-CoV-2 was detected in 27.7% (61/220) of diagnostic samples and in 5% (10/200) of screening samples. Overall, saliva test in diagnostic samples had a sensitivity of 88.5% (77.8-95.3%) and specificity of 98.1% (94.6-99.6%); in screening samples, the sensitivity was 90% (55.5-99.7%) and specificity 100% (98.1-100%). Our data suggests that the Fluidigm Advanta Dx RT-PCR saliva-based assay may be a reliable diagnostic tool for COVID-19 diagnosis in symptomatic individuals and screening asymptomatic healthcare workers.

Pearsonema (syn. Capillaria) plica is a nematode that resides in the urinary bladder of canids, felids and mustelids (definitive hosts) and is classified in the same class as Trichuris spp. Epidemiological and clinical data on Pearsonema plica infection in domestic animals are limited. The nematode has an indirect lifecycle that involves earthworms as intermediate hosts. A six-year-old crossbred dog from Greece, presented a history of intermittent pollakiuria and hematuria. At urine analysis, P. plica eggs were found in the urine sediment. The dog was successfully treated with a double dose of milbemycin. To the best of our knowledge this is the first case of urinary capillariasis diagnosed in a domestic animal in Greece., (Copyright © 2021. Published by Elsevier B.V.)

The aim of this study was to evaluate the performance of the new automated system Alfred 60 AST which is based on light scattering technology for rapid susceptibility testing directly from positive blood cultures as well as its applicability in the routine laboratory workflow. We evaluated 176 significant episodes of bacteremia due to 92 Gram-negative and 84 Gram-positive bacteria. The antimicrobial agents tested were ceftriaxone, ciprofloxacin, gentamicin, meropenem, piperacillin-tazobactam, and colistin for Gram negatives and cefoxitin, vancomycin, linezolid, and daptomycin for Gram positives. Concordance assessment was performed in comparison with our routine method, Vitek2 (bioMérieux). Discrepancies were resolved with MICRONAUT-S (Merlin) or E-test (bioMérieux). Out of 690 susceptibility determinations, 94.05% showed categorical agreement (CA) with the routine method and this percentage increased to 94.49 after discrepancy analysis. There were 1.45% very major errors, 3.33% major errors, and 1.16% minor errors (decreased to 1.45, 3.04, and 1.01 after discrepancy analysis). The CA for most of the antibiotics was above 90% except for daptomycin for Gram positives (87.30%) and ceftriaxone for Gram negatives (88.23%). The concordance was slightly better for Gram negative than for Gram-positive bacteria (94.30 versus 93.70%, respectively). The total turnaround time for a complete Alfred 60 AST result was 6-6.5h. The evaluated method gave rapid and reliable results in a few hours, versus 48h for the conventional one. Implementing this technology in routine workflow allows clinicians to optimize the treatment on the same day of blood culture positivity with potential positive clinical benefits and impact on antibiotic stewardship.

Introduction: Prompt detection of antibiotic resistance genes in healthcare institutions is of utmost importance in tackling the spread of multi-drug resistant micro-organisms. We evaluated the Antimicrobial Resistance (AMR) Direct Flow Chip Kit versus phenotypic screening assays for rectal and nasopharyngeal specimens upon ICU admission., Methods: A total of 184 dual specimens (92 rectal and 92 nasopharyngeal swabs) from 92 patients were collected from 11/2017 to 8/2018. All swabs were subjected to both AMR and phenotypic tests according to their origin. The degree of agreement of the two methods was assessed by the kappa coefficient., Results: The kappa coefficient showed perfect agreement for MRSA, ESBLs, oxacillinases and vancomycin resistance genes (1.000, p<0.01) and very good agreement for mecA-positive CoNS, KPC-carbapenemases and metallo-beta-lactamases (0.870, p<0.01; 0.864, p<0.01; and 0.912, p<0.01, respectively)., Conclusion: The AMR Direct Flow Chip Kit is a useful alternative to phenotypic testing for rapid detection of resistance markers., (Copyright © 2020 Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. Published by Elsevier España, S.L.U. All rights reserved.)

Ceftaroline is a novel cephalosporin able to bind to and inhibit PBP2a, and thus active against methicillin-resistant Staphylococcus aureus. In the present study we assessed the in vitro activity of ceftaroline and comparators against a large sample of methicillin-resistant and methicillin-susceptible S. aureus isolates collected at our hospital. Overall, both MRSA and MSSA isolates in our study were sensitive to ceftaroline, even though the MIC range was higher for MRSAs (0.12-2 mg/L against ≤0.06-0.5 mg/L for MSSAs). Our results indicate that ceftaroline may be considered a reliable alternative for the treatment of MRSA.

Bacterial carbapenem resistance, especially when mediated by transferable carbapenemases, is of important public health concern. An increased number of metallo-β-lactamase (MBL)-producing Klebsiella pneumoniae strains isolated in a tertiary hospital in Thessaloniki, Greece, called for further genetic investigation.The study included 29 non-repetitive carbapenem resistant K. pneumoniae isolates phenotypically characterized as MBL-producers collected in a tertiary hospital in Greece. The isolates were screened for the detection of carbapenemase genes (K. pneumoniae carbapenemase (blaKPC), Verona-integron-encoded MBL-1 (blaVIM-1), imipenemase (blaIMP), oxacillinase-48 (blaOXA-48) and New Delhi MBL (blaNDM)). The genetic relationship of the isolates was determined by Random Amplified Polymorphic DNA (RAPD) analysis. The whole genome sequences (WGS) from two NDM-positive K. pneumoniae isolates were further characterized.The presence of New Delhi MBL (blaNDM) gene was confirmed in all K. pneumoniae isolates, while blaKPC and blaVIM-1 genes were co-detected in one and two isolates, respectively. The RAPD analysis showed that the isolates were clustered into two groups. The whole genome sequence analysis of two K. pneumoniae isolates revealed that they belonged to the sequence type 11, they carried the blaNDM-1 gene, and exhibited differences in the number and type of the plasmids and the resistant genes.All MBL-producing K. pneumoniae isolates of the study harbored a blaNDM gene, while WGS analysis revealed genetic diversity in resistance genes. Continuous surveillance is needed to detect the emergence of new clones in a hospital setting, while application of antimicrobial stewardship is the only way to reduce the spread of multi-resistant bacteria.

The present study evaluated the carbapenem resistance mechanisms of Klebsiella pneumoniae strains isolated in two Greek tertiary teaching hospitals and their susceptibility to currently used and novel antimicrobial agents.Forty-seven carbapenem resistant K. pneumoniae strains were collected in G. Papanikolaou and Ippokrateio hospital of Thessaloniki between 2016 and 2018. Strain identification and antimicrobial susceptibility was conducted by Vitek 2 system (Biomérieux France). Susceptibility against new antimicrobial agents was examined by disk diffusion method. Polymerase chain reaction (PCR) was used to detect blaKPC, blaVIM, blaNDM and blaOXA-48 genes.The meropenem-EDTA and meropenem-boronic acid synergy test performed on the 24 K. pneumoniae strains demonstrated that 8 (33.3%) yielded positive for metallo-beta-lactamases (MBL) and 16 (66.6%) for K. pneumonia carbapenemases (KPC) production. Colistin demonstrated the highest in vitro activity (87.7%) among the 47 K. pneumoniae strains followed by gentamicin (76.5%) and tigecycline (51%). Among new antibiotics ceftazidime/avibactam showed the highest sensitivity (76.6%) in all strains followed by eravacycline (66.6%). The blaKPC gene was present in 30 strains (63.8%), the blaNDM in 11 (23.4%) and the blaVIM in 6 (12.8%). The blaOXA-48 gene was not detected.Well established antimicrobial agents such as colistin, gentamicin and tigecycline and novel antibiotics like ceftazidime/avibactam and eravacycline can be reliable options for the treatment of invasive infections caused by carbapenem-resistant K. pneumoniae.

Carbapenem-resistant Enterobacterales have become a major public-health issue worldwide. Here we report an outbreak caused by a clonal multidrug-resistant Proteus mirabilis strain producing VIM-4 metallo-β-lactamase (MBL) and TEM-2 β-lactamase in a Greek tertiary-care hospital. From July 2015 to February 2016, 27 imipenem-resistant P. mirabilis isolates were recovered from 14 patients hospitalised in two intensive care units (ICUs) and the internal medicine department in AHEPA University Hospital, Thessaloniki. The isolates were either susceptible or resistant to meropenem and were resistant to all remaining β-lactams except aztreonam. Phenotypic and molecular analysis revealed that all of the isolates harboured a bla VIM-4 MBL gene. Resistome analysis of a representative isolate showed the presence of an IncQ1 plasmid harbouring the bla VIM-4 carbapenemase and bla TEM-2 β-lactamase genes among resistance genes coding for resistance to β-lactams, aminoglycosides, trimethoprim, sulfonamides and lincosamides. Genotyping by pulsed-field electrophoresis (PFGE) revealed that the isolates were epidemiologically related. After recovery of the index carbapenemase-producing P. mirabilis clinical isolate, infection control measures were intensified in the affected departments. Rectal sampling for carbapenem-resistant bacteria was initiated on a weekly basis among patients admitted to the general ICU. The outbreak was finally interrupted 6 months later in February 2016. This is the first documentation of the bla VIM-4 MBL gene in P. mirabilis as well as the first hospital outbreak caused by a MBL-producing P. mirabilis strain., Competing Interests: Declaration of Competing Interest None declared., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Staphylococcus lugdunensis is considered more pathogenic than other coagulase-negative Staphylococci (CoNS), with its virulence resembling that of Staphylococcus aureus. We report a retrospective study of all S. lugdunensis infection cases during a 3.5-year period in a large tertiary university hospital in Greece. S.lugdunensis was susceptible to most tested antibiotics, although a high resistance percentage was found to clindamycin (27%) and erythromycin (25%). The susceptibility rate to penicillin was 49%, much lower than previously reported elsewhere, indicating that penicillin may not be an optimal treatment choice for S. lugdunensis infections in our region.

Objectives: Greece is endemic for KPC-encoding Klebsiella pneumoniae; however, until now, reports have referred only to hospital isolates. In this study, seven KPC-encoding K. pneumoniae isolated in private laboratories from non-hospitalised patients were characterised., Methods: Whole-genome sequencing was performed on an Illumina MiniSeq Sequencing System. Multilocus sequence typing (MLST) was performed using a BLAST-based approach, and antimicrobial resistance genes and plasmid replicons were identified using ResFinder and PlasmidFinder, respectively. The Rapid Annotation using Subsystem Technology (RAST) v.2.0 server was used for genome annotation of virulence, pathogenesis and defence genes., Results: Six isolates belonged to the major MLST sequence type 258 (ST258) and one to ST39. The resistome included genes encoding resistance mechanisms to β-lactams, aminoglycosides, quinolones, sulfonamides, trimethoprim, fosfomycin and phenicols, conferring multidrug-resistant phenotypes. Moreover, various genes involved in virulence, pathogenesis and defence have been identified., Conclusions: It is highly probable that these isolates were acquired during previous hospitalisation in Greek hospitals. The presence of KPC-encoding K. pneumoniae in non-hospitalised patients is alarming, although it is not yet possible to assess its actual impact., (Copyright © 2019 International Society for Antimicrobial Chemotherapy. Published by Elsevier Ltd. All rights reserved.)

Nintedanib is a tyrosine kinase inhibitor approved for the treatment of idiopathic pulmonary fibrosis (IPF). In a retrospective, real-world study across seven Greek hospitals, we evaluated the effectiveness and safety of nintedanib in routine clinical practice. Patients diagnosed with IPF, as per guideline criteria or multidisciplinary diagnosis, received nintedanib between January 2013 and January 2018. We evaluated 244 patients: mean±sd age 71.8±7.5 years, 79.1% male, 45.1% current smokers and 33.1% ex-smokers at treatment initiation. At baseline, predicted forced vital capacity (FVC) was 73.3±20.7% and predicted diffusing capacity of the lungs for carbon monoxide ( D LCO ) was 42.6±16.7%. On average, patients spent 23.6±15.0 months on nintedanib. At 3 years, 78 patients had died, equating to a 3-year survival rate of 59.4% (unaffected by treatment discontinuation or dose reduction). FVC% pred and D LCO % pred were largely stable at 3 years, with no significant difference from baseline (FVC 73.3±20.7% pred versus 78±20.1% pred, p=0.074; D LCO 42.6±16.7% pred versus 40.4±18.1% pred, p=0.334). Of the 244 patients, 55.7% reported an adverse event. Gastrointestinal events were the most common (173 (77.2%) out of 224 total events) and 45.0% of patients experienced diarrhoea. Only 32 (13.1%) patients had to permanently discontinue nintedanib due to an adverse event. This real-world study shows a 3-year survival rate of 59.4% and a low discontinuation rate due to adverse events. Our experience is consistent with previous findings in clinical trials of nintedanib in IPF., Competing Interests: Conflict of interest: K. Antoniou reports grants, personal fees, nonfinancial support and other support from BI Hellas during the conduct of the study, and other from Roche outside the submitted work. Conflict of interest: K. Markopoulou reports grants, personal fees, nonfinancial support and other support from BI Hellas during the conduct of the study, and other from Roche outside the submitted work. Conflict of interest: A. Tzouvelekis reports grants, personal fees, nonfinancial support and other support from BI Hellas during the conduct of the study, and other from Roche outside the submitted work. In addition, Dr Tzouvelekis has a patent on inhaled or aerosolised delivery of thyroid hormone to the lung as a novel therapeutic agent in fibrotic lung diseases issued to Yale University. Conflict of interest: A. Trachalaki has nothing to disclose. Conflict of interest: E. Vasarmidi has nothing to disclose. Conflict of interest: J. Organtzis has nothing to disclose. Conflict of interest: V. Tzilas has nothing to disclose. Conflict of interest: E. Bouros has nothing to disclose. Conflict of interest: G. Kounti reports grants, personal fees, nonfinancial support and other support from BI Hellas during the conduct of the study, and other from Roche outside the submitted work. Conflict of interest: C. Rampiadou reports grants, personal fees, nonfinancial support and other support from BI Hellas during the conduct of the study, and other from Roche outside the submitted work. Conflict of interest: S-C. Kotoulas has nothing to disclose. Conflict of interest: F. Bardaka has nothing to disclose. Conflict of interest: E. Bibaki has nothing to disclose. Conflict of interest: E. Fouka reports travel grants, lecture fees and consultation fees from Boehringer Ingelheim and GlaxoSmithKline, and travel grants and lecture fees from Roche Pharma, AstraZeneca, Novartis, Chiesi, Innovis and Elpen outside the submitted work. Conflict of interest: G. Meletis reports grants, personal fees, nonfinancial support and other support from BI Hellas during the conduct of the study. Conflict of interest: S. Tryfon has nothing to disclose. Conflict of interest: Z. Daniil reports grants, personal fees, nonfinancial support and other support from BI Hellas during the conduct of the study, and other from Roche outside the submitted work. Conflict of interest: D. Papakosta reports financial support for research, travel grants and lecture fees from Boehringer Ingelheim and Roche, outside the submitted work. Conflict of interest: D. Bouros reports research support, personal fees for lectures and advisory boards, and travel grants from Boehringer Ingelheim, and personal fees for lectures and advisory boards, and travel grants from Roche, outside the submitted work., (Copyright ©ERS 2020.)

Objectives: Klebsiella pneumoniae is a well-known pathogen frequently implicated in serious life-threatening nosocomial infections. Here we present a K. pneumoniae isolate (AHEPA1046) co-harbouring bla NDM-1 and bla OXA-48 isolated from a blood sample of an inpatient in Thessaloniki, Greece., Methods: Whole-genome sequencing (WGS) was performed using an Illumina MiniSeq Sequencing System. Multilocus sequence typing (MLST) was performed using a BLAST-based approach, and antimicrobial resistance genes and plasmid replicons were identified by ResFinder and PlasmidFinder, respectively. The Rapid Annotation using Subsystem Technology (RAST) v.2.0 server was used for genome annotation., Results: WGS analysis revealed the complete resistome of K. pneumoniae AHEPA1046. The strain harboured bla NDM-1 and bla OXA-48 together with 16 additional antimicrobial resistance genes and was resistant to carbapenems, aminoglycosides, quinolones, macrolides, tetracyclines, trimethoprim, fosfomycin and phenicols. Moreover, it was classified as ST11., Conclusion: This is the first report of a K. pneumoniae clinical isolate from Greece co-producing NDM-1 and OXA-48 carbapenemases and is one of a few reported worldwide., (Copyright © 2019 International Society for Antimicrobial Chemotherapy. Published by Elsevier Ltd. All rights reserved.)

The emergence and spread of NDM-1-encoding Klebsiella pneumoniae is causing worldwide concern, whereas a second epicenter of their dissemination after the Indian subcontinent is thought to be located in the Balkans. In this study, the complete genome sequencing of an NDM-1-producing ST11 K. pneumoniae isolated in a private laboratory in Greece is presented. The genome sequencing was performed on Illumina MiniSeq. Multilocus Sequence Typing was determined using a BLAST-based approach whereas antimicrobial resistance genes and plasmid replicons were identified by ResFinder and PlasmidFinder respectively. The capsular serotype was determined by the nucleotide sequence of the wzc gene. The Rapid Annotation System Technology server v2.0 was used for genome annotation. The isolate was classified to Sequence Type 11 and to the K24 capsular serotype. Its genome consisted of 5,549,974 bp with a G + C content of 57.26%. The resistome included 16 antibiotic resistance genes, 12 located in plasmids and 4 in the chromosome. The whole genome sequence of the isolate has been deposited at GenBank to serve as future reference in the study of the epidemiology and antibiotic resistance mechanisms of carbapenemase-producing Enterobacteriaceae.

Objectives: The emergence and spread of transferable β-lactamases among Enterobacteriaceae is a major problem both to human and veterinary medicine and is an important contributing factor to the development of multidrug-resistant bacterial isolates. In the present study, whole-genome sequencing of a Klebsiella pneumoniae isolate (LKP817909) resistant to first- and second-generation cephalosporins and non-susceptible to fluoroquinolones, isolated from a urine sample of a hospitalised dog, was performed., Methods: Genome sequencing was performed on an Illumina MiniSeq Sequencing System. Multilocus sequence typing (MLST) was performed using a BLAST-based approach, whereas antimicrobial resistance genes and plasmid replicons were identified by ResFinder and PlasmidFinder, respectively. The Rapid Annotation using Subsystem Technology (RAST) server v.2.0 was used for genome annotation., Results: Data analyses revealed the complete resistome of isolate LKP817909, which included the cefotaximase-München-11 (CTX-M-11) extended-spectrum β-lactamase together with 11 other resistance genes. Ten resistance genes were located on plasmids and two on the chromosome., Conclusions: To the best of our knowledge, this is the first detection of a CTX-M-11-producing K. pneumoniae isolated from a canine. The whole genome sequence of the isolate has been deposited at GenBank to serve as a future reference., (Copyright © 2018 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.)

The global spread of carbapenemase-encoding genes among Gram-negative nosocomial pathogens has led to the revival of polymyxins. Colistin and polymyxin B, despite their serious adverse effects, have become last resort treatment options for multi- or even extensively-drug-resistant bacterial infections due to Klebsiella pneumoniae, Acinetobacter baumannii and Pseudomonas aeruginosa. Their use, however, has been followed by an increase in polymyxin resistance rates and the spread of transferable resistance genes limiting further the treatment options and contributing to the emergence of pan-drug-resistance. In the present review, the to-date known polymyxin resistance mechanisms, as well as patents related to polymyxin resistance, are discussed., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Aerococcus urinae is a microorganism responsible for urinary tract and blood stream infections which are rarely reported in clinical practice. However, it has been proposed that the infrequency of such reports may be partially due to difficulties related to pathogen identification. We present here a case of an elderly male patient with urinary tract infection where A. urinae was initially not identified by a private microbiology laboratory. Our report highlights the need to consider A. urinae as a causative agent of urinary tract infections because if not identified and properly treated it may lead to endocarditis or septicemia.

Carbapenem resistance, mainly among Gram-negative pathogens, is an ongoing public-health problem of global dimensions. This type of antimicrobial resistance, especially when mediated by transferable carbapenemase-encoding genes, is spreading rapidly causing serious outbreaks and dramatically limiting treatment options. In this article, important key points related to carbapenem resistance are reviewed and future perspectives are discussed.

In 2010 the Hellenic center for disease control and prevention launched the "Prokroustes" nationwide action plan to tackle the increasing rates of carbapenem resistance among gram-negative nosocomial pathogens. In the present report, data from a Greek tertiary-care hospital are presented three years after the adoption of the infection control measures. Carbapenem resistance rates have been contained for Klebsiella pneumoniae and Acinetobacter baumannii but not for Pseudomonas aeruginosa. More worryingly, in accordance with their overuse against carbapenem-resistant bacteria, resistance rates to colistin and tigecycline have risen significantly.

The spread of carbapenemase-producing bacteria constitutes a worldwide problem of major importance to public health. The detection of these enzymes is often reported by most European countries and infection control measures are implemented in order to limit their dissemination. Despite the strong evidence indicating the Balkans as a reservoir for carbapenemase-encoding genes, especially for NDM-1, data published in the English literature seem to be scarce for many of the Balkan countries. We systematically reviewed studies on carbapenemase-producing bacteria from each country of the Balkan Peninsula and correlated them with foreign reports on carbapenemase detection due to patient transfer from the Balkans to the rest of Europe.

Beta-lactam antibiotics constitute the agents most widely used against bacterial infections worldwide. Hence, the emergence and spread of beta-lactam hydrolyzing enzymes among pathogenic bacteria has proved to be one of the major medical issues especially when these enzymes inactivate carbapenems together with other beta-lactams. The prompt detection of beta-lactamases is important for therapeutic and epidemiological purposes therefore, various appropriate methodologies have been developed throughout the last decades. In the present overview, recent patents related to the detection of beta-lactamases and especially of carbapenemases are discussed.

Acinetobacter baumannii clinical isolates have the ability to survive in the hospital niche for prolonged time periods and to develop resistance against multiple antimicrobial agents. Therefore, A. baumannii has emerged as an important cause of nosocomial outbreaks worldwide, especially in critical-care environments such as intensive care units. In the present communication, we report a multi-drug-resistant A. baumannii outbreak that occurred in an orthopedics department in Greece after the admission of a patient previously hospitalized in the intensive care unit of a Greek tertiary care hospital. Despite the implementation of infection control measures, 29 patients were infected, significantly raising their hospitalization periods and treatment costs. Interestingly, the outbreak was put under control after the department's previously programmed relocation., Competing Interests: The authors declare no conflict of interest.

Aim: The aim of this preliminary study was to investigate the in vitro effect of "non-antibiotic" trimebutine against reference strains Staphylococcus aureus ATCC 29213, ATCC 25923, Escherichia coli ATCC 25922, ATCC 35218, Pseudomonas aeruginosa ATCC 27853 and Enterococcus faecalis ATCC 29212; microbiota that are potentially involved in the pathophysiology of post-infectious functional gastrointestinal disorders., Methods: Trimebutine activity was assessed by the broth microdilution method according to Clinical and Laboratory Standards Institute recommendations against reference strains S. aureus ATCC 29213 and ATCC 25923, E. coli ATCC 25922 and ATCC 35218, P. aeruginosa ATCC 27853 and E. faecalis ATCC 29212. Bactericidal activity of the compound was determined by spreading a 10 μL aliquot on Mueller-Hinton agar from each dilution showing non-visible growth. All tests were carried out in triplicate., Results: Trimebutine was active against all strains tested presenting with MIC ranging from 1024 to 4000 mg/L. MIC and MBC were similar for E. coli ATCC 25922 and P. aeruginosa ATCC 27853 whereas for Gram-positive isolates and E. coli ATCC 35218 the MBC was higher., Conclusions: We demonstrated the in vitro bacteriostatic/bactericidal activity of trimebutine against bacteria frequently colonizing the gastrointestinal tract and potentially involved in human gastrointestinal infections that might trigger post-infectious functional gastrointestinal disorders.

A Klebsiella pneumoniae clinical isolate resistant to imipenem was recovered from a wound sample. The patient, a 57-year-old man, underwent a surgical resection of small bowel and sigmoid colon and was treated with multiple courses of antimicrobials. PCR analysis revealed that the clinical isolate was carrying simultaneously bla(VIM-1), bla(KPC-2), bla(SHV) and bla(TEM) genes. The concomitant presence of these genes is alarming and poses therapeutic as well as infection control problems.