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Background Aztreonam/avibactam is one of the last therapeutic options for treating infections caused by NDM-like-producing Enterobacterales. However, PBP3-modified and NDM-producing Escherichia coli strains that co-produce CMY-42 have been shown to be resistant to this drug combination. The aim of our study was to assess the in vitro activity of cefepime/taniborbactam and cefepime/zidebactam against such aztreonam/avibactam-resistant E. coli strains. Methods MIC values of aztreonam, aztreonam/avibactam, cefepime, cefepime/taniborbactam, cefepime/zidebactam and zidebactam alone were determined for 28 clinical aztreonam/avibactam-resistant E. coli isolates. Those isolates produced either NDM-5 (n = 24), NDM-4 (n = 2) or NDM-1 (n = 2), and they all co-produced CMY-42 (n = 28). They all harboured a four amino acid insertion in PBP-3 (Tyr-Arg-Ile-Asn or Tyr-Arg-Ile-Lys). Results All strains were resistant to aztreonam/avibactam and cefepime, as expected. The resistance rate to cefepime/taniborbactam was 100%, with MIC50 and MIC90 being at 16 mg/L and 64 mg/L, respectively. Conversely, all strains were susceptible to cefepime/zidebactam, with both MIC50 and MIC90 at 0.25 mg/L. Notably, all strains showed low MICs for zidebactam alone, with MIC50 and MIC90 at 0.5 mg/L and 1 mg/L. Conclusions Our data highlighted the excellent in vitro performance of the newly developed β-lactam/β-lactamase inhibitor combination cefepime/zidebactam against aztreonam/avibactam-resistant E. coli strains, suggesting that this combination could be considered as an efficient therapeutic option in this context. Our study also highlights the cross-resistance between acquired resistance to aztreonam/avibactam and the cefepime/taniborbactam combination.

We report in vivo development of cefiderocol (FDC) resistance among four sequential Pseudomonas aeruginosa clinical isolates ST244 recovered from a single patient, without exposure to FDC, which raises concern about the effectiveness of this novel drug. The first recovered P. aeruginosa isolate (P-01) was susceptible to FDC (2 μg/mL), albeit this MIC value was higher than that of a wild-type P. aeruginosa (0.12–0.25 μg/ml). The subsequent isolated strains (P-02, P-03, P-04) displayed increasing levels of FDC MICs (8, 16, and 64 μg/ml, respectively). Those isolates also showed variable and gradual increasing levels of resistance to most β-lactams tested in this study. Surprisingly, no acquired β-lactamase was identified in any of those isolates. Whole-genome sequence analysis suggested that this resistance was driven by multifactorial mechanisms including mutational changes in iron transporter proteins associated with FDC uptake, ampC gene overproduction, and mexAB-oprM overexpression. These findings highlight that a susceptibility testing to FDC must be performed prior to any prescription.

The in vitro activity of imipenem-relebactam, meropenem-vaborbactam, ceftazidime-avibactam, and cefiderocol was evaluated against both clinical and isogenic enterobacterial isolates producing carbapenemases of the SME, NmcA, FRI, and IMI types. Ceftazidime-avibactam and meropenem-vaborbactam showed the highest activity against all tested isolates; imipenem-relebactam showed only moderate activity.

Cefiderocol (FDC) and ceftazidime-avibactam (CZA) are among the latest generation of commercialized antibiotics against carbapenem-resistant Gram negatives. However, emergence of CZA resistance is being increasingly reported, involving different KPC variants in Enterobacterales. By analyzing two CZA-resistant KPC-3 clinical variants, KPC-41 and KPC-50, we showed that KPC-41, and to a lesser extent KPC-50, may also have an impact on susceptibility to FDC leading to a cross-resistance. This feature highlights that a susceptibility testing to FDC is mandatory prior any clinical use of FDC for treating infections due to KPC producers.

A multidrug-resistant (carbapenems, aztreonam + avibactam, and cefiderocol) ST167 Escherichia coli clinical isolate recovered from a patient hospitalized in Switzerland produced NDM-35 showing ca. 10-fold increased hydrolytic activity toward cefiderocol compared to NDM-1. The isolate co-produced a CMY-type β-lactamase, exhibited a four amino-acid insertion in PBP3, and possessed a truncated iron transporter CirA protein. Our study identified an association of unrelated resistance mechanisms leading to resistance to virtually all β-lactams in a high-risk E. coli clone.

A multidrug-resistant (carbapenems, aztreonam + avibactam, and cefiderocol) ST167 Escherichia coli clinical isolate recovered from a patient hospitalized in Switzerland produced NDM-35 showing ca. 10-fold increased hydrolytic activity toward cefiderocol compared to NDM-1. The isolate co-produced a CMY-type β-lactamase, exhibited a four amino-acid insertion in PBP3, and possessed a truncated iron transporter CirA protein.

Background Cefiderocol is among the latest generation of commercialized antibiotics against a large variety of MDR Gram-negative bacteria including carbapenem-resistant Enterobacterales and non-fermenters such as Pseudomonas aeruginosa and Acinetobacter baumannii. Cefiderocol susceptibility testing, a key element for implementing rapidly a cefiderocol-based treatment, might be still challenging. Objectives To develop a rapid culture-based test, Rapid Cefiderocol NP test, for the identification of cefiderocol resistance among MDR Enterobacterales. Methods The Rapid Cefiderocol NP test is based on glucose metabolization when bacterial growth occurs and the detection of bacterial growth in the presence of cefiderocol at 64 mg/L using iron-depleted CAMHB. Bacterial growth is visually detectable by a red-to-yellow colour change of red phenol, a pH indicator. A total of 74 clinical enterobacterial isolates from various clinical sources and of worldwide origin, among which 42 isolates were cefiderocol resistant, were used to evaluate the test performance. Results The sensitivity and specificity of the test were found to be 98% and 91%, respectively, by comparison with the reference broth microdilution (BMD) method. All positive results were obtained within 3 h after incubation at 35°C ± 2°C, that is a gain of time of ca. 18 h (1 day) compared with currently used techniques for susceptibility testing (BMD method). Conclusions This novel test is rapid, highly sensitive, specific, easily interpretable, and easy to implement in routine microbiology laboratories. Such a test may rapidly and accurately provide the information needed for the implementation of adequate cefiderocol-based treatment.

The increasing spread of carbapenem resistance is a serious global public health concern that negatively affects human and animal health. In this study we characterized the carbapenemase production...

The ability of broad-spectrum β-lactamases to reduce the susceptibility to ceftazidime-avibactam (CZA), ceftolozane-tazobactam (C/T), imipenem-relebactam, meropenem-vaborbactam, aztreonam-avibactam (AZA), and cefiderocol (FDC) was evaluated both in Pseudomonas aeruginosa and in Escherichia coli using isogenic backgrounds. Although metallo-β-lactamases conferred resistance in most cases, except for AZA, several clavulanic-acid-inhibited extended-spectrum β-lactamases (PER, BEL, SHV) had a significant impact on the susceptibility to CZA, C/T, and FDC.

The K.N.I.V.O Detection K-Set is a novel immunochromatographic test for detection of the 5 major carbapenemases (KPC, NDM, IMP, VIM, and OXA-48-like). This test is rapid, easy to perform, and shows a good specificity and sensitivity (96.8% and 100%, respectively), being suitable for microbiology laboratories together with biochemical rapid tests.

Objectives: The present work was conducted to investigate the nutritional profile of the highly consumed beef luncheon in Egypt. Besides, the potential health hazards associated with the con­sumption of luncheon were highlighted. Material and methods: A total of 60 beef luncheon samples were collected from Egyptian mar­kets. They were classified into three classes: A, B, and C based on their prices. The collected samples were examined for their chemical composition by determining moisture, protein, fat, ash, carbohydrate, and energy percentage. The contents of trace elements were also investigated. Results: The obtained findings showed a comprehensive dissimilarity in the chemical composition. According to the fresh weight base, moisture, protein, fat, ash, and carbohydrate fluctuated from 56.97 to 64.52, 3.50 to 16.10, 4.73 to 13.39, 3.30 to 3.51, and 11.32 to 27.44% w/w, respectively. The highest price class A Egyptian beef luncheon had more accepted nutritive value and dietary energy content. All the examined classes were low in calcium, potassium, zinc, and magnesium. The target hazard quotient indicated that the trace elements did not present any risks for con­sumers except for sodium. High phosphorous content and high phosphorus–protein ratio were observed in all categories which had a harmful health effect, hence named “new cholesterol.” Conclusion: Based on the information, this study is the leading work that deeply investigated the chemical composition of the Egyptian luncheon classes, and the obtained data could be beneficial to update the nutritional knowledge used by dietitians and the responsible for nutrition assess­ment and surveillance by the government. Adv. Vet. Anim. Res., 7(2): 299-307, June 2020

Polymyxins are becoming an added value for treating infections due to multidrug-resistant Pseudomonas aeruginosa. Therefore, the Rapid Polymyxin/Pseudomonas NP test was developed for the identification of polymyxin resistance in P. aeruginosa isolates. This test detects the bacterial growth in a medium containing a defined concentration of colistin. Formation of basic metabolites is visually detected by yellow to purple/violet color change of the bromocresol purple pH indicator. The test performance was evaluated by using 50 colistin-resistant and colistin-susceptible P. aeruginosa isolates, among which 10 were colistin resistant. The sensitivity and specificity of the test were found to be 100% (CI95 65.5-100%) and 95% (CI95 81.8-99.1%), respectively, by comparison with broth microdilution as the reference method. The Rapid Polymyxin/Pseudomonas NP test is easy to perform, specific and sensitive, allowing rapid visual observation of results without the requirement any special reading equipment, and results are obtained in 3 h.

A selective medium for screening fosfomycin (FOS)-resistant Enterobacterales was developed. Performances of this medium were first evaluated by using cultures of a collection of 84 enterobacterial clinical strains (42 FOS-susceptible and 42 FOS-resistant). The SuperFOS medium showed excellent sensitivity and specificity of detection (100%) in those conditions. Then, by testing spiked stool and spiked urine specimens, it revealed excellent performances, with lower limits of identification ranging from 101 to 102 CFU/ml. This screening medium allows easy and accurate detection of FOS-resistant isolates regardless of their resistance mechanisms.

A selective medium for screening fosfomycin (FOS)-resistant Enterobacterales was developed. Performances of this medium were first evaluated by using cultures of a collection of 84 enterobacterial clinical strains (42 FOS susceptible and 42 FOS resistant). The SuperFOS medium showed excellent sensitivity and specificity of detection (100%) in those conditions. Then, by testing spiked stool and spiked urine specimens, it revealed excellent performances, with lower limits of identification ranging from 10(1) to 10(2) CFU/ml. This screening medium allows easy and accurate detection of FOS-resistant isolates regardless of their resistance mechanisms.

Cefiderocol (FDC) is a recently developed siderophore cephalosporin showing excellent antibacterial activity against Gram-negative bacteria, including Acinetobacter baumannii . By investigating a series of A. baumannii clinical isolates with elevated MICs of FDC, we showed that PER-like β-lactamases and, to a lesser extent, NDM-like β-lactamases, significantly contributed to reduced susceptibility to that antibiotic.

Among the most common metallo-β-lactamases (MBL), the most clinically significant are the New Delhi Metallo-β-lactamases (NDM). NDM enzymes hydrolyze almost all β-lactams except aztreonam and are not inhibited by novel β-lactamase inhibitors such as avibactam (AVI), relebactam, and vaborbactam (1).….

Objectives This study describes the first draft genome sequence of a multidrug-resistant (MDR) Escherichia coli D-ST69 clinical isolate from Egypt carrying blaNDM-1 and blaOXA-244. Methods The strain was isolated in December 2014 from a wound pus swab of a male patient in the city of Kafr El-Sheikh using MacConkey agar containing 2 μg/mL meropenem. The strain was subjected to antimicrobial susceptibility testing, conjugation experiments, and whole-genome sequencing using an Illumina MiSeq platform. Results The draft genome of the strain (HR14_AS) was 5.08 Mbp in size containing a total of 90 contigs encoding 4677 predicted genes with an average G+C content of 50.7%. Strain HR14_AS belongs to sequence type 69 (ST69), phylogroup D and exhibits an MDR phenotype, with minimum inhibitory concentrations (MICs) of 64 μg/mL and 32 μg/mL for meropenem and doripenem, respectively. Multiple acquired antimicrobial resistance genes conferring resistance to macrolides [mdf(A)], fluoroquinolones [aac(6')-Ib-cr], quinolones (qnrS1), trimethoprim (dfrA14), β-lactams (blaNDM-1, blaOXA-244, blaCTX-M-15, blaOXA-9 and blaTEM-1B) and aminoglycosides [aac(3)-IId, aac(6')-Ib, aadA1 and aph(3')-VI] were detected. The blaOXA-244 and blaNDM-1 genes were located on the chromosome (Tn6237) and on an IncI1-type self-conjugative plasmid of >93 kb in size, respectively. Conclusions Here we report the first draft genome sequence of a MDR E. coli D-ST69 isolate carrying blaNDM-1 and blaOXA-244. Besides clonal expansion of the E. coli ST38 pandemic clone, this study further identified that the spread of OXA-244-producing E. coli could be related to mobilisation of the IS1R-made composite transposon (Tn6237) carrying blaOXA-244.

The Rapid ResaImipenem/Acinetobacter NP test was developed for the identification of carbapenem resistance among Acinetobacter baumannii isolates. The principle of this test is based on the reduction of resazurin (a viability colorant) by metabolically active bacterial cells, hence detecting bacterial growth, in the presence of a defined concentration of imipenem chosen to be slightly above that defining imipenem resistance (6 μg/ml). The bacterial growth is visually detected by a color change from blue (resazurin) to purple or pink (resorufin product). A total of 110 A. baumannii isolates, among which 61 were imipenem-resistant, were used to evaluate the test performance. The sensitivity and specificity of the test were found to be 100%, by comparison with broth microdilution taken as the reference standard method. The Rapid ResaImipenem/Acinetobacter NP test is highly specific and sensitive, and easy to implement in routine microbiology laboratories and results are obtained within 2 h 30. It does not require any specific equipment.

The industrial version of SuperCAZ/AVI® medium developed for screening CAZ/AVI resistant Gram-negative isolates has been evaluated here using a collection of 87 well-characterized clinical isolates of worldwide origin. In addition, testing was performed by spiking stools with a series of resistant and susceptible isolates. In those conditions, the SuperCAZ/AVI® medium exhibited a sensitivity and specificity of 100 %, down to the lower limit of detection of 101 to 102 CFU/ml. The SuperCAZ/AVI® medium is a sensitive and specific screening medium for detection of CZA-resistant bacteria regardless of their resistance mechanisms.

A homemade and culture-based test, relying on the visual detection of the reduction of the resazurin reagent (a cell viability indicator), has been developed for the rapid detection of polymyxin resistance in Acinetobacter baumannii. Here, we evaluated the industrial version of this test, the RapidResa Polymyxin Acinetobacter NP® test (Liofilchem, Italy). A well-characterized panel of 68 clinical A. baumannii strains (36 polymyxin-susceptible, 26 polymyxin-resistant A. baumannii, and 6 colistin-heteroresistant isolates) of worldwide origin was tested. All the colistin-susceptible A. baumannii isolates gave negative results according to the RapidResa Polymyxin Acinetobacter NP® test, except for a single isolate that gave a false-positive result. Out of the 26 colistin-resistant A. baumannii strains, 25 were correctly identified as colistin resistant using the RapidResa Polymyxin Acinetobacter NP® test. Only a single colistin-resistant A. baumannii strain gave a false-negative result. Additionally, the six colistin-heteroresistant isolates tested gave positive results. Altogether, the sensitivity and the specificity of the test were found to be 96% and 97%, respectively. The turn-around-time of this easy-to-perform test was 3-4h, which showed excellent reliability for identification of polymyxin resistance in A. baumannii. The RapidResa Polymyxin Acinetobacter NP® test allows a rapid differentiation between polymyxin-susceptible and -resistant A. baumannii isolates, which may contribute to optimization of the use of polymyxins for treating infections due to multidrug-resistant A. baumannii.

A series of clinical NDM-5-producing E. coli isolates obtained from two surveillance networks of carbapenem-producing Enterobacterales from 2018-2019, namely Switzerland (NARA) and Germany (SurvCARE), were analyzed. The 33 NDM-5- producing E. coli isolates were highly resistant to β-lactams including the novel ß- lactam/ß-lactamase inhibitor combinations (ceftazidime/avibactam, imipenem/relebactam and meropenem/vaborbactam), and remained susceptible to fosfomycin, colistin, and tigecycline. Those isolates were assigned to different sequence types (STs) and indicated a predominance of isolates exhibiting the sequence type (ST) ST167 in Switzerland and in Germany (n=10, phylogenetic group C), followed by ST405 (n=4, phylogenetic group E), ST1284 (n=4, phylogenetic group C) and ST361 (n=4, phylogenetic group C). The blaNDM-5 gene was predominantly present on an IncF-type plasmid (n=29), and to a lesser extent on the narrow-host range IncX3 plasmid (n=4). Sequence analyses of eight NDM-5 plasmids indicated that NDM-5-encoding F-type plasmids varied in size between 86 to 132 kb. The two IncX3 plasmids pCH8NDM5 and pD12NDM5 were 46 and 45 kb in size, respectively. The highly conserved blaNDM-5 genetic surrounding structures [ΔISAba125-blaNDM- 5-bleMBL-trpT-dsbD-IS26] of both the F-type and IncX3 plasmids suggested a common genetic origin. The emergence of the NDM-5 carbapenemase was evidenced in particular for the E. coli ST167 clone, which is a successful epidemic clone, known to be associated to both multi-resistance and virulence traits and is therefore of high public health concern. The occurrence of clonally related NDM-5- producing E. coli isolates in Switzerland and Germany further indicates the international spread of this multidrug-resistant superbug at least throughout Europe.

Volume 58, no. 9, e00965-20, 2020, [https://doi.org/10.1128/JCM.00965-20][1]. This article was published on 24 August 2020 with a CC BY 4.0 copyright line (“Copyright © 2020 Sadek et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0

Volume 58, no. 9, e00932-20, 2020, [https://doi.org/10.1128/JCM.00932-20][1]. This article was published on 24 August 2020 with a CC BY 4.0 copyright line (“Copyright © 2020 Nordmann et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0

Metallo-β-lactamase (MBL)-producing Escherichia coli isolates resistant to the newly developed β-lactam/β-lactamase inhibitor drug combination aztreonam-avibactam (ATM-AVI) have been reported. Here, we analyzed a series of 118 clinical MBL- producing E. coli isolates of various geographical origins for susceptibility to ATM-AVI. The nature of the PBP3 protein sequence and the occurrence of blaCMY genes for susceptibility to ATM-AVI were investigated. We showed here that elevated MICs of ATM-AVI among MBL-producing E. coli isolates resulted from a combination of different features, including modification of PBP3 protein sequence through specific amino acid insertions and production of CMY-type enzymes, particularly, CMY-42. We showed here that those insertions identified in the PBP3 sequence are not considered the unique basis of resistance to ATM-AVI, but they significantly contribute to it.

The NitroSpeed-Carba NP test was used to rapidly detect and discriminate between the different types of carbapenemases (classes A, B, and D) within 30 minutes among a collection of 202 Pseudomonas sp. strains (mostly Pseudomonas aeruginosa). A total of 99 carbapenemase–(including enzymes exhibiting weak carbapenemase activity such as several Guyana Extended-Spectrum (GES)-ß-lactamases) and 103 non-carbapenemase producers were tested, and the overall specificity and sensitivity were 100% and 99%, respectively. The NitroSpeed-Carba NP test is a rapid, specific, sensitive, and easy-to-implement technique for identification of carbapenemase- producing Pseudomonas spp.

The SuperCAZ/AVI medium was developed for screening ceftazidime-avibactam (CZA) resistance among Gram-negative bacteria (Enterobacterales and Pseudomonas aeruginosa). It was evaluated using 50 CZA-susceptible and 42 CZA-resistant Gram-negative isolates. Its sensitivity and specificity of detection were 100%. Excellent performance of the medium was also observed by testing spiked stools, with the lower limit of detection ranging from 101 to 102 CFU/ml., The SuperCAZ/AVI medium was developed for screening ceftazidime-avibactam (CZA) resistance among Gram-negative bacteria (Enterobacterales and Pseudomonas aeruginosa). It was evaluated using 50 CZA-susceptible and 42 CZA-resistant Gram-negative isolates. Its sensitivity and specificity of detection were 100%. Excellent performance of the medium was also observed by testing spiked stools, with the lower limit of detection ranging from 101 to 102 CFU/ml. This screening medium provides the opportunity to detect CZA-resistant isolates regardless of their resistance mechanisms.

A biochemical test (NitroSpeed-Carba NP test) was developed to identify carbapenemase production in Enterobacterales and to discriminate between the different types of clinically significant carbapenemases (Ambler classes A, B, and D). It is based on two main features, namely, the hydrolysis by all β-lactamases, including carbapenemases of the nitrocefin substrate, and the capacity of ertapenem to prevent this hydrolysis for all β-lactamases except carbapenemases. Specific carbapenemase inhibitors of class A (avibactam, vaborbactam), class B (dipicolinic acid), and class D (avibactam) were used to inhibit the nitrocefin hydrolysis and to allow the identification of the carbapenemase types with a turnaround time of ca., A biochemical test (NitroSpeed-Carba NP test) was developed to identify carbapenemase production in Enterobacterales and to discriminate between the different types of clinically significant carbapenemases (Ambler classes A, B, and D). It is based on two main features, namely, the hydrolysis by all β-lactamases, including carbapenemases of the nitrocefin substrate, and the capacity of ertapenem to prevent this hydrolysis for all β-lactamases except carbapenemases. Specific carbapenemase inhibitors of class A (avibactam, vaborbactam), class B (dipicolinic acid), and class D (avibactam) were used to inhibit the nitrocefin hydrolysis and to allow the identification of the carbapenemase types with a turnaround time of ca. 30 min. The test was evaluated with a collection of 248 clinical enterobacterial isolates, including 148 carbapenemase producers and 100 non-carbapenemase producers. Its overall sensitivity and specificity were 100% and 97%, respectively, including detection of all types of OXA-48-like carbapenemases. For the detection of the carbapenemase type, including strains that produce double carbapenemases, the sensitivity was 100%, 97%, and 100% for the detection of classes A, B, and D, respectively. This easy-to-implement test may contribute to optimization of the choice of the β-lactam/β-lactamase inhibitor combinations for treating infection due to carbapenemase producers.

We describe here the complete genome sequence of an Enterobacter hormaechei ST279 coharbouring blaVIM-1 and mcr-9 recovered from uncooked beef patty in June 2017, Egypt. The tested isolate was resistant to carbapenem but susceptible to colistin (minimum inhibitory concentration (MIC), 0.5 &mu, g/mL). The antimicrobial susceptibility profile and conjugation experiments were performed. The entire genome was sequenced by the Illumina MiniSeq and Oxford Nanopore methods. The blaVIM-1 and mcr-9 genes are carried on the same IncHI2/pMLST1 plasmid, pMS37a (Size of 270.9 kb). The mcr-9 gene was located within the physical boundaries demarcated by two insertion elements IS903 (upstream) and IS1 (downstream) but did not possess the downstream regulatory genes (qseC/qseB) which regulate the expression of mcr-9. Therefore, the mcr-9 might be silently disseminated among carbapenem-resistant Enterobacterales. In addition to blaVIM-1 and mcr-9, plasmid pMS37a harbored various antibiotic resistance genes including aac(6&rsquo, )-Il, &Delta, aadA22, aac(6&rsquo, )-Ib-cr, sul1, dfrA1 and tetA. To the best of our knowledge, this is the first report of a blaVIM-1 and mcr-9-coharbouring E. hormaechei isolate of food origin worldwide. The identification of a multidrug-resistant VIM-1 and mcr-9 positive Enterobacter hormaechei isolate from food is worrisome as retail meat and meat products could serve as a vehicle for these MDR bacteria, which could be transferred between animals and humans through the food chain. It further highlights that Enterobacterales co-producing MCR and carbapenemases being found in the food chain indeed correspond to a One-Health issue, highlighting the need for serious steps to prevent their further dissemination.

The increasing spread of carbapenem resistance is a serious global public health concern that negatively affects human and animal health. In this study we characterized the carbapenemase production in gram-negative bacteria isolated from different meat and meat products in Egypt. Phenotypic and genotypic susceptibility testing were investigated. Two

The Revogene® Carba C assay is a real-time polymerase chain reaction–based assay that runs on the microfluidic Revogene platform. It was recently designed for the detection of genes encoding the 5 major carbapenemases (NDM, VIM, IMP, KPC, and OXA-48) from various Gram-negatives. A total of 145 clinical Gram-negative strains (96 carbapenemase producers and 49 non–carbapenemase producers) were tested. The overall sensitivity and specificity were 100%. All strains co-producing double carbapenemases have been correctly detected. All non–carbapenemase producers and nontargeted carbapenemase producers gave a negative result. The sample preparation was easy to handle, taking around 5 to 10 min per isolate, with a run time of approximately 70 min. This assay is a rapid, easy-to-perform, reliable tool to detect the most common carbapenemases, with excellent sensitivity and specificity regardless of the host bacteria. Given its user friendliness, simplicity, and short time to result, the Revogene® Carba C assay is suitable for microbiology laboratories.

PurposeThis study aims to investigate the nutrient composition and cholesterol content of most popular fast foods sold in Qena city, Egypt with reference to theirtransfatty acids contents.Design/methodology/approachTotal 80 samples of fast foods were collected from various fast food restaurants in Qena city, Egypt. The samples were investigated for their nutritive quality by measuring moisture, protein, fat, ash, carbohydrate and energy content as described by the association of analytical chemists. The cholesterol content was determined using a spectrophotometer, whiletransfatty acids were analyzed by gas chromatography using standard methods.FindingsThe study revealed a wide variation in the composition and nutritive value. On a fresh weight basis, moisture, protein, fat, carbohydrate and ash ranged from 45.9-55.0 (P< 0.001), 15.0-22.3 (P< 0.001), 13.1-15.7 (P= 0.034), 6.0-16.2 (P< 0.001) and 2.2-3.5 (P< 0.001), %w/w, respectively. The fast foods were very energy dense with calorie content varying from 236.5-281.1 (P< 0.001), Kcal/100 g. The mean values of cholesterol were between 15-17 mg/100 g (P= 0.398). The findings showed that fast foods had high totaltransfatty acids content exceeding the Danish legal limit of 2 g/100 g of fat being in the range of 3.5 to 11.3 g per 100 g fat (P= 0.379). Elaidic acid is the predominant trans-isomer in industrially producedtransfatty acids. The concentrations of elaidic acid were high in all samples examined (P= 0.942).Originality/valueThis study has established the fact that fast foods are associated with high total energy intake, high intake of fat, trans fat and a higher proportion of calories being derived from total fat and trans-fat. Therefore, it is recommended to limit the intake of fast food.

Objectives The aim of this study was to investigate the occurrence of plasmid-encoded colistin resistance among Gram-negative bacteria isolated from meat and meat products in Egypt and to report the draft genome sequence of anmcr-1/IncI2-carrying multidrug-resistant (MDR) Escherichia coli B1:ST101 isolate. Methods A total of 128 colistin-resistant strains were isolated from various meat and meat product samples in different cities in Egypt. Multiplex PCR screening for plasmid-mediated colistin resistance genes was performed. Whole-genome sequencing was performed using an Illumina NextSeq platform and the genome was assembled using CLC Genomics Workbench 7.5.1. Results A singlemcr-1-positive MDR E. coli strain was isolated from beef sausages. The genome size of the E. coli strain was calculated at 5 044 715 bp, with a total of 226 contigs and a G + C content of 50.5%. The strain belonged to ST101 (phylogroup B1). The mcr-1 gene was located on an IncI2-type self-conjugative plasmid of 64.6 kb in size. The strain showed a MDR phenotype, with a colistin MIC of 4 mg/L. A large number of acquired antimicrobial resistance genes was identified, including genes encoding resistance to colistin (mcr-1), β-lactams (blaTEM-1), phenicols (floR), trimethoprim (dfrA12), aminoglycosides [aac(3)-IIa, aph(3”)-Ib and aadA2], macrolides (mphA and mdfA), tetracyclines (tetA), sulfonamides (sul1 and sul2) and quinolones (qnrS1). Conclusion Here we report the first draft genome sequence of anmcr-1/IncI2-carrying MDR E. coli B1:ST101 isolated from beef sausage in Egypt. This study highlights the potential role played by food products in the spread of colistin resistance to humans.

This study compared the impact of different approaches, namely, nonenrichment, nonselective enrichment, and selective (antibiotic-containing) enrichment steps, for detecting extended-spectrum β-lactamase producing Enterobacterales (ESBL-E), carbapenemase-producing Enterobacterales (CPE), polymyxin-resistant Enterobacterales (PMR-E), and vancomycin- resistant enterococci (VRE) from spiked stools. The use of a nonselective 18-h enrichment broth culture significantly improved the recovery rate of all types of resistant bacteria after their plating onto selective media. In addition, the detection of ESBL-E, CPE, PMR-E, and VRE was further improved when using an enrichment step using antibiotic-supplemented broths respectively supplemented with cefotaxime (0.1 μg/mL), ertapenem (0.1 μg/mL), colistin (0.5 μg/mL), and vancomycin (1 μg/mL). Therefore, we showed here that a screening strategy based on a selective broth enrichment step significantly contributes to an increased rate of detection of multidrug-resistant bacteria, which may be crucial in term of improvement of infection control.