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This study was conducted in Bejaia, Algeria, to determine the presence of Salmonella in fresh watermelon ( n  = 105), soil ( n  = 23), and irrigation water samples ( n  = 17) collected from two different farms. After isolation, antimicrobial susceptibility testing, serotype determination, multilocus sequence typing, antimicrobial resistance genes detection, and whole genome sequencing were performed. Twenty watermelon samples (19%) were contaminated with Salmonella , but none were found in the soil or irrigation water. Among the 20 Salmonella isolates, 2 serovars were identified ( Salmonella Liverpool and Salmonella Anatum), belonging to sequence types ST1959 and ST64, respectively. Ten Salmonella isolates showed significant resistance to nalidixic acid, ofloxacin, and ciprofloxacin but were susceptible to all other antibiotics. The coexistence of point mutations ( parC :p.T57S) in Quinolone Resistance-Determining Regions and the qnrB19 gene may contribute to quinolone resistance. The study identified 164 virulence genes in the Salmonella isolates. Our study found Salmonella in fresh watermelon during the preharvest season in Bejaia, Algeria. Our study indicates a relatively high prevalence of Salmonella on watermelon samples before harvest. Although we cannot directly compare our results with previous studies, it is crucial to recognize that the absence of comprehensive comparative data underscores the need for further research and surveillance.

Nontyphoidal Salmonella presents a significant threat to animal and human health as a food-borne infectious agent. This study focused on the characterization of Salmonella isolates obtained from fresh animal products consumed in Bejaia, Algeria. In total, 495 beef products, 600 chicken products, and 355 dairy products were collected from retail outlets and slaughterhouses in the region. Out of the 1450 samples collected, a Salmonella positivity rate of 3.5% (51/1450) was observed. Traditional Algerian sausages exhibited the highest prevalence of Salmonella contamination (14.1%, 36/256). Fifteen different serotypes were identified, with S. Kentucky ( n = 12), S. Anatum ( n = 11), and S. Bredeney ( n = 9) being the most prevalent. Antimicrobial susceptibility testing showed resistance to diverse antibiotics, particularly against tetracycline, ampicillin, nalidixic acid, and sulphonamides. Whole-genome sequencing conducted on 21 isolates enabled the comparison of phylogenetic links between isolates. We notably identified clones circulating across the region in different locations and food types, suggesting contamination at the early stages of the food chain (in the herd or slaughterhouses) that disseminated to numerous butcher shops in various cities. We also identified acquired antibiotic resistance genes and point mutations in the quinolone resistance-determining region genes, contributing to the observed resistance patterns. This study describes the genomic and phylogenetic characteristics of several Salmonella clones found in diverse food samples in Algeria. It suggests potential transmission dynamics that could better understand Salmonella 's contamination routes.

This study investigated Salmonella spp. in wild animals in Algeria, focusing on their prevalence, serotypes, antibiotic resistance, and virulence profiles. From fecal samples collected between May 2021 and June 2022, 1.9% showed Salmonella shedding. The identified serotypes included S. Bredeney, S. Enteritidis, S. Altona, and S. Virchow. Except for S. Altona, all isolates were resistant to quinolones, with S. Bredeney strains, exhibiting multidrug resistance. Whole-genome sequencing revealed various resistance genes and mutations in gyrA or parC genes. Additionally, plasmids IncX1 and ColpVC were detected in several isolates. A comprehensive analysis identified 201 virulence genes. These findings contribute to understanding Salmonella in wild animal populations and their potential impact on public health., (© 2024. EcoHealth Alliance.)

An increase of carbapenemase-producing Bacteroides fragilis infections is observed. To detect such a resistance in B. fragilis, several tests exist that are expensive or show poor sensitivity and specificity. Therefore, we upgraded the Anaerobic Carbapenem Inactivation Method (Ana-CIM) to easily screen for carbapenemase-producing B. fragilis. The presence of carbapenemase cfiA gene was identified in 50 B. fragilis isolates by PCR. We modified the Ana-CIM by (1) increasing the bacterial inoculum, and (2) measuring the differences in diameter between the negative control and the testing disc. We correctly classified the cfiA-negative and positive isolates and could define a cut-off of positivity at 2 mm. Our modified Ana-CIM allowed to correctly discriminate the 31 cfiA-positive with meropenem MICs ranging from 1 to > 32 µg/mL. We anticipate that our modified Ana-CIM could be used in most clinical laboratories to easily screen for carbapenemase-producing B. fragilis, even at low levels., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Bacteria within biofilms may be exposed to sub-minimum inhibitory concentrations (sub-MICs) of antibiotics. Cell-to-cell contact within biofilms facilitates horizontal gene transfers and favors induction of the SOS response. Altogether, it participates in the emergence of antibiotic resistance. Aminoglycosides at sub-MICs can induce the SOS response through NO accumulation in E. coli carrying the small plasmid with the quinolone resistance qnrD gene (pDIJ09-518a). In this study, we show that in E. coli pDIJ09-518a, the SOS response triggered by sub-MICs of aminoglycosides has important consequences, promoting genetic rearrangement in class 1 integrons and biofilm formation. We found that the integrase expression was increased in E. coli carrying pDIJ09-518a in the presence of tobramycin, which was not observed for the WT isogenic strain that did not carry the qnrD -plasmid. Moreover, we showed that biofilm production was significantly increased in E. coli WT/pDIJ09-518a compared to the WT strain. However, such a higher production was decreased when the Hmp-NO detoxification pathway was fully functional by overexpressing Hmp. Our results showing that a qnrD -plasmid can promote biofilm formation in E. coli and potentiate the acquisition and spread of resistance determinants for other antibiotics complicate the attempts to counteract antibiotic resistance and prevention of biofilm development even further. We anticipate that our findings emphasize the complex challenges that will impact the decisions about antibiotic stewardship, and other decisions related to retaining antibiotics as effective drugs and the development of new drugs.

The plasmid-mediated quinolone resistance (PMQR) genes have been shown to promote high-level bacterial resistance to fluoroquinolone antibiotics, potentially leading to clinical treatment failures. In Escherichia coli , sub-minimum inhibitory concentrations (sub-MICs) of the widely used fluoroquinolones are known to induce the SOS response. Interestingly, the expression of several PMQR qnr genes is controlled by the SOS master regulator, LexA. During the characterization of a small qnrD -plasmid carried in E. coli, we observed that the aminoglycosides become able to induce the SOS response in this species, thus leading to the elevated transcription of qnrD . Our findings show that the induction of the SOS response is due to nitric oxide (NO) accumulation in the presence of sub-MIC of aminoglycosides. We demonstrated that the NO accumulation is driven by two plasmid genes, ORF3 and ORF4, whose products act at two levels. ORF3 encodes a putative flavin adenine dinucleotide ( FAD )-binding oxidoreductase which helps NO synthesis, while ORF4 codes for a putative fumarate and nitrate reductase ( FNR )-type transcription factor, related to an O 2 -responsive regulator of hmp expression, able to repress the Hmp-mediated NO detoxification pathway of E. coli . Thus, this discovery, that other major classes of antibiotics may induce the SOS response could have worthwhile implications for antibiotic stewardship efforts in preventing the emergence of resistance., Competing Interests: AB, DS, AM, GP, ZB, TJ, MP, SG, FB, SV, SM, Cd, DM, TG No competing interests declared, (© 2022, Babosan et al.)

Chronic obstructive pulmonary disease (COPD) is a chronic inflammatory lung disease characterized by airflow limitation. This chronic respiratory disease represents the third leading cause of death worldwide. Alteration of the airway microbiota has been reported to be associated with exacerbation frequency in COPD, but its role on the symptoms in patients at stable state is still incompletely described. This study aimed to determine whether bacteria isolated in sputum can be associated with the clinical features of COPD patients within stable state. Our study highlights, for the first time, that altered microbiota with Enterobacterales is associated with pejorative clinical symptoms in stable COPD patients. The airway microbiota of 38 patients was analyzed using an extended culture approach and mass spectrometry identification. Cluster analysis by principal coordinate analysis of the bacterial communities showed that the patients could be classified into three distinct clusters in our cohort. The clusters showed no differences in proportions of the phylum, but one of them was associated with a high prevalence of Enterobacterales (71.4% in cluster 1 vs. 0% in cluster 3), loss of microbiota diversity, and higher bacterial load (10 7 vs. 10 5 CFU/ml, respectively) and characterized by predominant cough and impact on mental health. These novel findings, supported by further studies, could lead to modifying the processing of COPD sputum in the everyday practice of clinical microbiology laboratories., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Muggeo, Perotin, Brisebarre, Dury, Dormoy, Launois, Ancel, Mulette, de Champs, Deslée and Guillard.)

Objectives: The emergence and worldwide spread of carbapenemase-producing Enterobacterales (CPE) is a great public-health concern. This study aimed to screen for the presence of CPE isolates from Barbary deer in Akfadou Forest, Béjaïa (Algeria)., Methods: Faecal samples (n=39) were obtained from Barbary deer in Akfadou Forest between March-June 2018. Whole-genome sequencing (WGS) was performed to characterise one representative strain of Klebsiella pneumoniae. Data analysis was performed using online tools., Results: A total of 13 carbapenem-resistantK. pneumoniae isolates were obtained. The isolates showed an identical antimicrobial resistance pattern and were susceptible to colistin and fosfomycin. WGS analysis revealed the complete resistome of K. pneumoniae strain CF21, including bla NDM-1 , bla CTX-M-15 , bla SHV-182 , bla DHA-1 , bla OXA-1 , aac(3)-IIa, aac(3)-IId, aac(6')-Ib-cr, rmtC, sul1, qnrB9, fosA, tetA, dfrA14, catA2, catB3 and mphA. Multilocus sequence typing (MLST) analysis assigned this strain to the international clone ST11. Plasmid analysis showed that this K. pneumoniae strain possesses five different plasmids including IncA/C2, IncFIA(HI1), IncFIB(K), IncFII(K) and ColRNAI., Conclusion: This study reports a multidrug-resistantK. pneumoniae strain recovered from Barbary deer in Algeria and confirms that wild animals could serve as a reservoir of antimicrobial resistance genes., (Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Background: Quinolone resistance (QR) is one component of the MDR emerging in Escherichia coli and is of particular concern given the widespread use of fluoroquinolones., Objectives: To characterize the QR phenotypes and genotypes in E. coli responsible for bloodstream infections and to propose molecular determinants that could be targeted to predict ciprofloxacin resistance., Methods: E. coli isolates from blood cultures in three French hospitals were studied for quinolone MICs and characterization of genotypic QR determinants (QRg)., Results: Among 507 isolates tested for MICs, 148 (29.2%) were resistant to quinolones based on EUCAST breakpoints and 143 (28.2%) harboured at least one QRg. QRg were mainly mutations in the QRDR (138 isolates, 27.2%), with 55.8% of these isolates carrying at least three QRDR mutations. gyrA mutations predominated (92.8%) followed by parC (61.6%), parE (32.6%) and gyrB (1.4%) mutations. Only 4.7% of the isolates harboured a plasmid-mediated quinolone resistance (PMQR) gene: aac(6')-Ib-cr (60.0%) or qnr (qnrS, qnrB) (32.0%). For the first time in France, we reported the qepA4 allele of the plasmid-encoded efflux pump QepA. Only five isolates carried PMQR without a QRDR mutation. The positive predictive value (PPV) for ciprofloxacin resistance was 100% for any QRg and 99.2% for gyrA mutations specifically., Conclusions: QR observed in E. coli isolates involved in bloodstream infections is still mainly due to QRDR mutations, especially at codons GyrA83/87, which could be used as a molecular target to rapidly detect resistance., (© The Author(s) 2020. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Carbapenem resistance in Enterobacteriaceae has become an increasingly worrying threat. So far, no epidemiological data regarding NDM-producing enterobacterial isolates has been available on these strains in West Africa. The aim of this study was to seek for carbapenemase-producing Enterobacteriaceae clinical strains isolated in Bamako Teaching Hospital in Mali. Of 50 strains isolated between May 2016 and September 2016, we found a ST448 E. coli harbouring an IncX3 plasmid with bla NDM-5 embedded in the Δ ISAba125-ble MBL structure. This study reports the first description of NDM-5 in Mali isolated in an undescribed ST E. coli in West Africa.

Objective: The aim of this study was to investigate the resistance mechanisms of carbapenem-resistant Enterobacteriaceae clinical strains recovered from Al Thawra University Hospital, Sana'a, Yemen. Methods: A total of 27 isolates showing decreased susceptibility to carbapenems were obtained from different clinical specimens in Al Thawra Hospital, Sana'a, Yemen. Strains were identified by Matrix Assisted Laser Desorption Ionization Time-Of-Flight spectroscopy. Susceptibility to antibiotics was determined by the disk diffusion method on Mueller Hinton agar. Carbapenemases-encoding genes, extended-spectrum β-lactamases (ESBLs), and plasmid-mediated quinolone resistance (PMQR) genes were screened by PCR. Bacterial isolates were typed by multilocus sequence typing (MLST). Results: Carbapenemase genes detection and sequencing showed that 18 (66.7%) isolates were Klebsiella pneumoniae (NDM-1, n  = 13; NDM-1 + OXA-48, n  = 3; OXA-48, n  = 1; OXA-232, n  = 1), 6 (22.2%) were Escherichia coli (NDM-5, n  = 3; OXA-181, n  = 2; OXA-48, n  = 1), and 3 (11.1%) were Enterobacter cloacae (NDM-1, n  = 1; OXA-181, n  = 2). In addition the ESBL gene bla CTX-M-15 was detected in 14 K. pneumoniae and 2 E. coli isolates, and the bla CTX-M-216 was found in 1 E. coli isolate. Fifteen isolates were PMQR positive including qnrB1 ( n  = 1), qnrS1 ( n  = 5), qnrS4 ( n  = 2), and aac-(6')-Ib-cr ( n  = 7). The MLST typing showed a diversity of sequence type (ST) clones including Escherichia coli ST410 (3), ST448 (2), and ST648; Enterobacter cloacae ST78 and ST270; and Klebsiella pneumoniae ST395 (2), ST309, ST23, ST35, ST1728, ST15, ST231, and ST1428. Conclusion: This study reports the first description of OXA-48-like-producing Enterobacteriaceae and NDM-5 enzymes in E. coli in Yemen.

Background: Pathogens involved in healthcare-associated infections can quickly spread in the environment, particularly to frequently touched surfaces, which can be reservoirs for pathogens., Aim: The purpose of this study was to investigate naturally occurring bacterial contamination on touch surfaces in five French long-term care facilities and to compare bacterial populations recovered from copper and control surfaces., Methods: More than 1300 surfaces were sampled. The collected bacteria were identified to obtain a global view of the cultivable bacterial populations colonizing touch surfaces. Haemolytic colonies and putative pathogens were also screened using specific agar plates and then identified with matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry. In total, more than 3400 colonies were analysed., Findings: Staphylococcus and Micrococcus were the two predominant genera present on touch surfaces, respectively occurring on 51.8% and 48.0% of control surfaces. In these facilities with relatively low bioburden, copper surfaces efficiently reduced the occurrence frequencies of three genera: Staphylococcus, Streptococcus and Roseomonas. Pathogenic species such as Staphylococcus aureus, Enterococcus faecalis and E. faecium were observed in very few samples. In addition, meticillin-resistant S. aureus was observed on five control surfaces and one copper surface., Conclusion: Contamination of healthcare facilities touch surfaces can be the source for the spread of bacteria through the institution. This in situ study shows that the frequency of the contamination as well as the specific bacterial population bioburden is reduced on copper alloy surfaces., (Copyright © 2019 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.)

Objective: We aimed to study the efficacy of copper as an antimicrobial agent by comparing incidence rates during outbreaks in areas equipped vs not equipped with copper surfaces in a long-term facility for dependent older adults (nursing home)., Design: Prospective observational pilot study in a nursing home., Setting and Participant: All persons resident in the nursing home belonging to Reims University Hospital, from February 1, 2015 to June 30, 2016, were included., Methods: Incidence rates for health care-related infections during outbreaks occurring during the study period were compared between the wing that was equipped and the wing that was not equipped with copper surfaces. Results are expressed as relative risks (RRs) and 95% confidence intervals (95% CIs)., Results: During the study period, 556 residents were included; average age was 85.4 ± 9.2 years, and 76% were women. Four outbreaks occurred during the study period: 1 influenza, 1 keratoconjunctivitis, and 2 gastroenteritis outbreaks. The risk of hand-transmitted health care-associated infection was significantly lower in the area equipped with copper surfaces (RR 0.3, 95% CI 0.1-0.5)., Conclusions and Implications: In our study, copper was shown to reduce the incidence of hand-transmitted health care-associated infections and could represent a relatively simple measure to help prevent HAIs in nursing homes., (Copyright © 2019 AMDA – The Society for Post-Acute and Long-Term Care Medicine. Published by Elsevier Inc. All rights reserved.)

Cardiobacterium hominis est un bacille à Gram négatif responsable d'endocardites infectieuses, principalement chez les patients atteints de pathologies cardiaques ou porteurs de valves. L'identification de cette bactérie est souvent complexe et peut être la cause d'un diagnostic et d'une prise en charge tardifs, source de complications cardiaques. Cet article présente la prise en charge d'une endocardite infectieuse associée à un sepsis à Cardiobacterium hominis, les difficultés d'identification de cette bactérie, ainsi qu'une revue de la littérature sur les infections dues à cette bactérie.

The aim of this study was to screen for the presence of carbapenemase-producing Enterobacteriaceae (CPE) isolates from bat guano in Bejaia, Algeria. Guano samples ( n  = 110) were collected in Aokas's cave, Bejaia, Algeria, between March and May 2016. Samples were plated on MacConkey agar supplemented with ertapenem (0.5 mg/L) and vancomycin (32 mg/L). The isolates were identified and antimicrobial susceptibility was determined using disk diffusion method. Carbapenemase, extended spectrum β-lactamases, plasmid-mediated AmpC, and plasmid-mediated quinolone resistance genes were studied using PCR and sequencing. Clonal relatedness was studied using multilocus sequence typing (MLST). Two CPE isolates were identified as Klebsiella pneumoniae . PCR and sequencing identified the bla OXA-48 in one K. pneumoniae strain (CS34) and bla KPC-3 in the other strain (CS63). K. pneumoniae CS63 was found to carry bla TEM-1 and aac(6')-Ib genes. The MLST showed that K. pneumoniae CS63 was assigned to ST512, whereas K. pneumoniae CS34 belonged to ST1878. This is the first description of CPE from bats' guano.

Introduction: The spread of carbapenemase-producing Enterobacteriaceae (CPE) represents a major public health issue. Methods allowing rapid detection of carbapenemases in developing countries are therefore urgently needed. In the current study, we developed a new in-house medium for the rapid detection of CPE isolates, especially OXA-48 producers., Methodology: A panel of 144 clinical strains previously characterized was tested on in-house Carba MTL-broth medium using four different concentrations of ertapenem (0.5 to 2 mg/L), and compared to chromID® OXA-48 and chromID® CARBA (BioMérieux) media., Results: Comparative evaluation of the Carba MTL-broth with chromID® OXA-48 and chromID® CARBA showed that chromID® OXA-48 and Carba MTL-broth had the highest sensitivity for detection of OXA-48 producers (93.9% and 100%, respectively) comparatively to chromID® CARBA (21.2%). The chromID® OXA-48 had the highest specificity (100%), as compared to the Carba MTL-broth (65.5%) and chromID® CARBA (84.4%) for the detection of OXA-48 producers., Conclusions: The in-house Carba MTL-broth developed in this study is sensitive, inexpensive, an easy-to-use phenotypic method for the detection of OXA-48-producing enterobacteria. Given the burden of pan-drug resistance, its implementation in the microbiology laboratory of developing countries could be a useful tool for rapid detection of these bacteria., Competing Interests: No Conflict of Interest is declared, (Copyright (c) 2019 Assia Mairi, Abdelaziz Touati, Alix Pantel, Catherine Dunyach-Remy, Albert Sotto, Christophe De Champs, Jean-Philippe Lavigne.)

In the healthcare environment, microorganisms' cross-transmission between inanimate surfaces and patients or healthcare workers can lead to healthcare-associated infections. A recent interest has grown to create antimicrobial copper touch surfaces, in order to counteract microbial spread in the healthcare environment. For the first time, five French long-term care facilities were at 50% fitted with copper alloys door handles and handrails. Related to the environmental bacterial contamination, 1400 samples were carried out on copper and control surfaces over three years after copper installation. In addition, some copper door handles were taken from the different facilities, and their specific activity against methicillin-resistant S. aureus (MRSA) was tested in vitro. In comparison to control surfaces, copper door handles and handrails revealed significantly lower contamination levels. This difference was observed in the five long-term care facilities and it persists through the three years of the study. High and extreme levels of bacterial contamination were less frequent on copper surfaces. Although, the antibacterial activity of copper surfaces against MRSA was lowered after three years of regular use, it was still significant as compared to inert control surfaces. Therefore, copper containing surfaces are promising actors in the non-spreading of environmental bacterial contamination in healthcare facilities.

Objectives: Fluoroquinolones (FQs) are a potential treatment for infections caused by extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae that are susceptible to these agents., Methods: Owing to increasing non-susceptibility to carbapenems among Enterobacteriaceae, in this study FQ resistance mechanisms were characterised in 36 ertapenem-non-susceptible Klebsiella pneumoniae isolated from North-Eastern France in 2012. The population structure was described by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST)., Results: Among the 36 isolates, 13 (36%) carried a carbapenemase encoding-gene. Decreased expression of the OmpK35-encoding gene might be considered a major resistance determinant that could explain the non-susceptibility to carbapenems. The carbapenemase-producing isolates carried the well-known IncL pOXA-48a plasmid. All 36 K. pneumoniae isolates also harboured a FQ resistance determinant. The aac(6')-Ib-cr gene was the major plasmid-mediated quinolone resistance (PMQR) determinant found in K. pneumoniae (89%; 32/36). MLST identified five sequence types (STs), with the most common being ST395 (69%; 25/36), followed by ST147 (17%; 6/36). ST395 strains showed ertapenem minimum inhibitory concentrations (MICs) ranging from 0.75-32μg/mL. Klebsiella pneumoniae ST395 isolates did not show enhanced biofilm formation or environmental survival but showed higher chlorhexidine MICs compared with ST147 isolates., Conclusions: These findings showed that (i) K. pneumoniae ST395 carrying pOXA-48a has spread in North-Eastern France, (ii) aac(6')-Ib-cr is predominant in carbapenem-non-susceptible K. pneumoniae, (iii) K. pneumoniae ST395 is resistant to chlorhexidine and (iv) FQs as an alternative to β-lactams to treat ertapenem-non-susceptible K. pneumoniae are compromised., (Copyright © 2017 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.)

Introduction: The city of Villeurbanne has made the political choice to reduce social inequalities in health (SIH). The Public Health Department wanted to assess the health impact in the context of this project. The main objective was to analyse the links between urban development and health in order to identify the impact of the urban project on health and to make recommendations to reduce physical inactivity of children. Attention was focused on a neighbourhood of 6,000 inhabitants in the process of urban renewal. A high rate of adults on antidiabetic treatment was observed in this relatively deprived neighbourhood. School medicine also noted a high proportion of overweight children in schools in the neighbourhood, higher than in all schools of the city. The objective of this HIA was therefore to identify the impact of the renovation project on children's mobility and to make recommendations in order to combat their physical inactivity by encouraging spontaneous physical activity., Methods: The selected determinants were grouped into four categories: outdoor environment, use of open spaces, adapted sports facilities/equipment, roads. The evaluation was based on analysis of urban planning documents, environmental analysis reports, ad hoc surveys, individual interviews with the various partners and target actors. A bibliographic review confirmed the positive or negative impacts of various aspects of the project on health., Results: Although some aspects of the project had a positive impact on the health of the inhabitants, others have a negative impact on children's mobility, well-being and social capital. About 30 recommendations were proposed and prioritized according to their political support and their technical, economic and temporal feasibility., Discussion: The HIA created a dynamic cross-working between sectors other than health and provided an opportunity to educate and inform stakeholders on different concepts and their roles in promoting positive health behaviours. The HIA, as a tool to promote health, contributes to implementation of strategies designed to transform living conditions responsible for social inequalities in health.

Pasteurella multocida is rarely observed in human chronic infections. A Pasteurella multocida strain was isolated from a skin biopsy of chronic dermohypodermitis in a 21-year-old woman without an immunocompromised state. As this strain was viable one month after a cat scratch despite treatment by amoxicillin-clavulanic acid, we compared this strain's growth rate, amoxicillin Minimal Inhibitory and Bactericidal Concentrations (MIC and MBC), resistance to serum and ability to activate neutrophil granulocytes with those of control strains isolated during acute infections in humans without previous antibiotics exposure. This particular strain was not more resistant to serum and did not induce a lower phagocytic activity than control strains. It did not grow more slowly than control strains even after suboptimal exposure to amoxicillin. This particular strain was tolerant to amoxicillin but tolerance did not appear sufficient alone for the induction of a chronic infection in a host without an immunocompromised state.

A new phenotypic method for detecting carbapenemases has been adapted (assembling of two MAST® kits, including one that contains faropenem to which a temocillin disk has been added) then assessed using 101 bacterial strains (Enterobacteriaceae with assays on Pseudomonas aeruginosa and Acinetobacter baumannii) including 62 which produce genetically identified carbapenemases. Concerning Carbapenemase-Producing Enterobacteriaceae (CPE), there is 100% sensitivity for Klebsiella pneumoniae carbapenemase (KPC, Ambler class A) and OXA-48 (Ambler class D), and 91% for metallo-beta-lactamase (MBL, Ambler class B), with a 97% sensitivity for all carbapenemases, with a specificity of 100%. The test is also efficient for detecting Pseudomonas aeruginosa carbapenemases (sensitivity between 82 and 100% and 100% specificity). The major innovation is the combined use of faropenem and temocillin for reliable detection (excellent performance with 100% sensitivity and specificity) of OXA-48. This study has led to the development of a new algorithm to detect the different classes of carbapenemases, for first-line diagnosis, by combining this modified MAST® test with immunochromatographic methods and molecular biology techniques., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Carbapenemase-producing Klebsiella pneumoniae strains have emerged as a major problem for healthcare systems. The aim of this study was to determine the role and diversity of plasmids harboring carbapenemases encoding genes from a collection of K. pneumoniae isolates recovered between July 2011 and January 2012, with decreased susceptibility to carbapenems. Imipenem (IPM), ertapenem (ETP), meropenem (MEM), and doripenem (DOR) minimum inhibitory concentrations (MICs) were determined by E-test. Carbapenemase production was detected with the modified Hodge test. β-Lactamases encoding genes were amplified by PCR and sequenced. Plasmid incompatibility groups harbored by carbapenemases producers were investigated using the PCR-based replicon typing method and the clonal relationship of the isolates was investigated by pulse filed electrophoresis. IMP, ertapenem, meropenem, and doripenem MICs ranged between 0.25 and 16 mg/L. Carbapenemase activity was detected in 14 isolates. Two carbapenemases were identified: OXA-48 in 13 isolates and a new variant OXA-204 in 1 isolate, in combination with extended-spectrum β-lactamases, CTX-M-1, CTX-M-9, CTX-M-14, CTX-M-15, and VEB-8. One isolate produced CMY-2. OXA-48 and the new variant OXA-204 were confirmed as transferable plasmid encoded. The carbapenemase-producing K. pneumoniae harbored plasmids of the A/C, LVPK, and L/M replicon types. Thirteen different pulso types were observed. Three pairs of isolates showed a clonal relatedness. This diversity in β-lactamases, in pulso types and in plasmid content, shows the ability of OXA-type carbapenemase to disseminate. This is worrying for the control of the increase in antibiotic resistance frequency and necessitates that continuous investigations in the clinical setting remain a high priority to clarify the contribution of antimicrobial use into multiresistance bacterial dissemination.

Objectives: The aim of this study was to investigate carbapenemase-producing Enterobacteriaceae (CPE) in companion animals., Methods: Between October 2015 and April 2016, 533 rectal swabs were obtained from healthy and diseased pets in different cities in Algeria. Samples were plated on MacConkey agar supplemented with ertapenem (0.5mg/L). Isolates were identified by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF/MS). Antimicrobial susceptibility testing was performed by the disk diffusion method. Carbapenemase, plasmid-mediated AmpC (pAmpC), extended-spectrum β-lactamase (ESBL) and plasmid-mediated quinolone resistance genes were characterised by PCR. Plasmids were extracted by the Kieser extraction method and were analysed by PCR-based replicon typing (PBRT). The epidemiological relationship between Enterobacter cloacae isolates was determined by random amplified polymorphic DNA (RAPD) analysis and multilocus sequence typing (MLST)., Results: From 533 rectal swabs, 12 Enterobacteriaceae (2.3%), including 2 Escherichia coli, 2 Klebsiella pneumoniae and 8 E. cloacae, were recovered from selection plates. The 12 strains were resistant to amoxicillin/clavulanic acid, ticarcillin, piperacillin/tazobactam and ertapenem. All isolates were susceptible to aminoglycosides, imipenem and extended-spectrum cephalosporins. PCR and sequencing identified the bla OXA-48 gene in all isolates. qnrB1 was identified in all E. cloacae isolates. Plasmid analysis showed that the bla OXA-48 gene was localised on a 7-kb untypeable plasmid. RAPD analysis demonstrated the presence of the same profile pattern in the eight E. cloacae isolates. MLST analysis showed that the E. cloacae isolates belonged to ST527., Conclusions: This study reports for the first time the presence of CPE in horses and pet birds in the world., (Copyright © 2017 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.)

ComPath is an ongoing European programme dedicated to monitor antibiotic susceptibility of bacterial pathogens from diseased dogs and cats. The objective was to characterize determinants associated with quinolone resistance among 160 enrofloxacin non-wild type strains (100 Escherichia coli, 45 Proteus mirabilis, 14 Klebsiella pneumoniae, 1 Enterobacter cloacae) selected among 843 non-duplicate Enterobacteriaceae isolates collected in 12 European countries (2013-2014). These strains with non-wild type MICs of ≥0.25 mg/L, for P. mirabilis ≥0.5 mg/L, were screened for PMQR determinants (qnr, oqxAB, qepA and aac(6')-Ib-cr), and for QRDR mutations in gyrA, gyrB, parC and parE genes. Among them, 20% (32/160) carried at least one PMQR (18/32 qnrB, qnrS or qnrD, 10/32 aac(6')-Ib-cr and 13/32 oqxAB), and 80% (128/160) no PMQR. qnrB was detected in 3 E. coli, 2 K. pneumoniae and 1 E. cloacae strains; qnrS in 6 E. coli and 1 P. mirabilis and aac(6')-Ib-cr in 4 E. coli, 5 K. pneumoniae and 1 E. cloacae strains. All qnrD1 were detected in P. mirabilis. oqxAB was detected in 12/14 K. pneumoniae and 1 E. cloacae. No qepA genes were detected. From the 32 PMQR-positive strains, 10 showed enrofloxacin MICs ≤2 mg/L and 22 MICs ≥8 mg/L, the latter carrying 1-4 mutations in QRDR. For the 128 non-PMQR strains, 37 showed enrofloxacin MICs ≤2 mg/L with 0-2 QRDR mutations, and 91 MICs ≥4 mg/L carrying 1-4 QRDR mutations. In conclusion, qnr was the major PMQR and qnrD only detected in Proteeae. Mutations in QRDR play a markedly greater role in mediating fluoroquinolone resistance than PMQR., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Nocardia spp. est responsable d'infections opportunistes chez les patients immunodéprimés, principalement d'infections pulmonaires. Le système nerveux central est la localisation extra-pulmonaire la plus souvent retrouvée. Nous rapportons ici un cas d'infection cutanée à N. brasiliensis chez une patiente immunocompétente, ainsi qu'une revue de la littérature.

Objectives: The current study aimed to investigate extended-spectrum β-lactamase-producing Enterobacteriaceae (ESBL-E) fecal carriage in children with different cancers admitted in the pediatric oncology unit of Beni-Messous Hospital (Algiers, Algeria)., Materials and Methods: Rectal swabs from children with cancer were sampled from February 2012 to May 2013 within 48 hours following their admission. After species identification and detection of ESBL production by double-disk synergy test (DD test), antibiotic susceptibility was determined by the standard disk diffusion method. Antibiotic resistance genes, including bla genes and plasmid-mediated quinolone resistance (PMQR) genes, were investigated by polymerase chain reaction (PCR). The phylogenetic grouping of Escherichia coli strains was determined by PCR., Results: Of the 171 children studied, 93 (54%) were ESBL carriers. An antibiotic treatment for the last 3 months before admission (p = 0.01), hematological malignancies (p = 0.003), and death (p = 0.0003) were more frequent in the ESBL-E group than in the non-ESBL group. Multivariate analysis showed that hematological malignancies (odds ratio [OR]: 3.9; confidence interval [CI]: 1.1-14.1; p = 0.04) and ESBL-E carriage (OR: 6.2; CI: 1.7-22.00; p = 0.005) were two independent factors associated with increased risk of death. A total of 103 ESBL-E isolates were obtained. Klebsiella pneumoniae and E. coli isolates were the most frequently isolated. PCR amplification showed that all the isolates produced a CTX-M ESBL (CTX-M-15, CTX-M-14, and CTX-M-3). The PMQR genes detected were qnrB, qnrS, and aac(6')-Ib-cr. E. coli isolates were assigned to four major extraintestinal pathogenic E. coli phylogroups, including B2 and D., Conclusion: This study provides, for the first time, insight into epidemiology of the ESBL-E fecal carriage among children with cancer in Algeria.

Background: The impact of a minimum inhibitory concentration (MIC) of vancomycin ≥2 mg/L on mortality and the potential benefit of new antistaphylococcal treatments in coagulase-negative Staphylococcus (CoNS) bacteraemia remain unknown. We assessed the impact of vancomycin MIC on 30-day in-hospital mortality and identified factors independently associated with 30-day in-hospital mortality., Methods: All patients presenting significant CoNS bacteraemia in the university hospital of Reims, between 01 January 2008 and 31 December 2012, were included. Data were retrospectively extracted from the patient records. Vancomycin MIC was assessed using the E-test method, and antimicrobial susceptibility testing was performed in accordance with the recommendations of the Antibiogram Committee of the French Microbiology Society. Cox's Proportional Hazards model was used for multivariate analysis., Results: Two hundred and sixty-nine patients (mean age 61.2 ± 15.7 years) were included. Foreign material was present in 92% of patients and 78.4% of isolated methicillin-resistant strains had vancomycin MIC ≥2 mg/l. Thirty-day in-hospital mortality was 16%. There was no association between vancomycin MIC ≥2 mg/l and 30-day in-hospital mortality (adjusted Hazard Ratio (aHR) = .80, 95% confidence interval (95%CI) [.30-2.19], p = .67). Factors independently associated with 30-day in-hospital mortality were age ≥75 vs. ≤60 years (aHR =3.72, 95%CI [1.39-9.97], p = .009), absence of active antibiotic treatment (aHR =5.52, 95%CI [1.13-26.87], p = .03) and acute renal failure (aHR =4.45, 95%CI [2.08-9.56], p < .0001). Removal of an infected device had a protective effect against 30-day in-hospital mortality (aHR = .23, 95%CI [.11-.48], p < .0001)., Conclusions: These results suggest that CoNS bacteraemia should be managed by removal of the infected device and antibiotic treatment such as vancomycin.

Background: Carbapenems are frequently used as a last resort to treat infections caused by multidrug-resistant Gram-negative organisms, thus carbapenem-non-susceptible Enterobacteriaceae (CNSE) is an emerging health threat., Objectives: To assess risk factors and outcomes of CNSE carriage., Patients and Methods: We conducted a matched case-control study in six hospitals in North-Eastern France. The controls were patients harbouring carbapenem-susceptible Enterobacteriaceae. Fifty-five cases and 110 controls were included., Results: Most of the CNSE isolates were Enterobacter cloacae and Klebsiella pneumoniae . Carbapenemase production was observed in 40% of isolates and they produced OXA-48 only. CNSE carriage was significantly associated with recent antibiotic use ( P =  0.014), particularly carbapenems ( P =  0.03) and fluoroquinolones ( P =  0.016). A multivariate analysis using conditional logistic regression showed that the presence of concomitant infection(s) (OR: 9.83; 95% CI 3.04-21.39, P =  0.0031), nosocomial infections (OR: 7.84; 95% CI 2.00-12.54, P  =   0.0063) and a high age (OR: 1.07; 95% CI 1.01-1.06, P =  0.038) were independently associated with CNSE carriage. Moreover, patients infected with CNSE had worse outcomes: fewer resolved infections at 1 month ( P =  0.02), and they had a higher mortality rate ( P =  0.0004) and longer hospital stays ( P =  0.02)., Conclusions: We identified three independent risk factors for CNSE carriage as well as worse outcomes in infected patients in North-Eastern France. This highlights the importance of early detection of CNSE and the need for antimicrobial therapy re-evaluation after bacteriological analysis has been performed., (© The Author 2017. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

ComPath is a European monitoring programme dedicated to the collection of bacterial pathogens from diseased dogs and cats to determine their antibiotic susceptibility. The objective was to characterize genetic determinants associated with quinolone resistance among 69 enrofloxacin non-wild type strains selected among 604 non-duplicate Enterobacteriaceae isolates collected in 10EU countries from 2008 to 2010: quinolone resistance determining region (QRDR) and plasmid-mediated quinolone resistance (PMQR). Among them, 17% (12/69) carried at least one PMQR (9/12 qnrB, qnrS or qnrD and 4/12 aac(6')-Ib-cr) and 83% (57/69) no PMQR. All the Klebsiella pneumoniae isolates chromosomally carried oqxAB . No qepA genes were detected. Eight strains did not carry any mutations in QRDR (4 PMQR-positive and 4 PMQR-negative strains). From the 12 PMQR-positive strains, 4 showed enrofloxacin MICs≤2μg/mL, and 8 MICs≥8μg/mL (resistant). These latter strains carried 1-5 mutations in QRDR, including a ParE I529L mutation. qnrD was found in 2 Proteus mirabilis and the plasmids were similar to pDIJ09-518a previously described. For the 57 non-PMQR strains, 29 strains showed MICs≤2μg/mL (4 with no QRDR mutations, 21 with 1 mutation in GyrA, 4 with 2 mutations in GyrA) and 28 showed enrofloxacin MICs≥8μg/mL carrying at least 2 mutations in QRDR, including a ParE I529L mutation for 2 Escherichia coli strains with a total of 5 QRDR mutations. No GyrB mutations were found. qnr was the major PMQR and qnrD was only detected in Proteus spp. Twelve strains carried at least 4 mutations., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The aim of this study was to evaluate the rate of fecal carriage of Escherichia coli strains producing Extended-spectrum β-lactamases (ESBLs) and plasmid-mediated quinolone resistance (PMQR) isolated from healthy pets (dogs and cats) in Algeria. Fecal samples from 171 healthy pets (102 dogs and 69 cats) in one veterinary practice and private owners were included. After isolates identification, antibiotic susceptibility was determined by disk diffusion procedure. ESBL were detected by combination disk tests. PCR and sequencing were used to characterize genes encoding ESBLs and PMQR. Transfer of ESBL and PMQR genes was assessed by conjugation experiments. Phylogenetic groups of E. coli were determined by PCR. Of the 171 animals, 20 carried an ESBL producing E. coli giving a prevalence of ESBL fecal carriage of 11.7%. All isolates were susceptible to carbapenems, cefoxitin, piperacillin-tazobactam, amikacin and fosfomycine. For the rest of the tested β-lactams, susceptibility rates ranged from 35% to 70% for cefepime and amoxicillin-clavulanic acid respectively. Concerning the non-beta-lactams antibiotics, the rates of susceptibility ranged between 5% to trimethoprim and 95% for chloramphenicol. The beta-lactamase genes identified in E. coli isolates were blaCTX-M-15, blaCTX-M-1, blaSHV-12 and blaTEM-1. The PMQR determinants aac(6')-Ib-cr, qnrS1 and qnrB5 genes were identified in 15 isolates. Transconjugants were obtained for two isolates. Phylogenetic analysis showed that E. coli isolates belong to commensal phylogroups of A and B1. We reported here for the first time in Algeria ESBL and PMQR-producing E. coli in healthy cats and dogs., (Copyright © 2016 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

The emergence and worldwide spread of carbapenemase-producing Enterobacteriaceae is of great concern to public health. The aim of this study was to investigate the occurrence of carbapenemase-producing Escherichia coli in companion animals in Algeria. Two hundred fecal samples were obtained from healthy and diseased dogs and cats in one veterinary office and private owners in Bejaia city, Algeria, during November 2014 to March 2015. Isolates were screened by polymerase chain reaction for the presence of carbapenemase, acquired plasmidic AmpC (pAmpC) and extended-spectrum beta-lactamase genes. Five carbapenemase-producing E. coli isolates were detected including four OXA-48-producing isolates and one isolate producing NDM-5. Coexpression of ESBL and pAmpC genes was observed in these isolates. Phylogenetic grouping revealed that these isolates belonged to A and D phylogroups. The results of this study show that carbapenemase-producing E. coli spread to the companion animals in Algeria.

Erysipelothrix rhusiopathiae is mostly isolated in swine causing erysipelas. Human invasive infections due to E. rhusiopathiae remain poorly described and interestingly bacteraemia associated with endocarditis are a source of ineffective empirical antibiotherapy. We report a case of sepsis without endocarditis due to E. rhusiopathiae and a review of the literature.

The aim of this study was to determine the origin of virulence and multiresistance of a Klebsiella pneumoniae isolate from an abdominal wound infection of a patient with a gunshot injury in the thoracoabdominal region. The isolate was identified using biochemical tests and Phoenix™ automated system and was confirmed by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF/MS). MICs of each antibiotic were determined by Etest. Screening for carbapenemase production was performed by the modified Hodge test and was confirmed by PCR amplification. Virulence factors were also studied. Plasmid replicon typing was used to classify Incompatibility (Inc) plasmids harbouring the resistance genes. The transferability of each plasmid was determined by conjugation using Escherichia coli J53. Finally, multilocus sequence typing (MLST) was performed to determine the ST of the strain. The bacterial isolate was identified as K. pneumoniae and was named KPM2, carrying entB, ybtS, mrkD and ycfM virulence genes, but it did not overexpress OqxAB. Isolate KPM2 belonged to ST147 and was classified as resistant to all of the tested antibiotics with MICs above the clinical breakpoints. These resistances were due to production of OXA-48, CMY-2, TEM-1, CTX-M-15 and VEB-8 β-lactamases. Genetic and molecular studies showed that blaOXA-48 was embedded in transposon Tn1999.2 and was carried by a conjugative IncL/M plasmid of ca. 60kb; blaVEB-8 was harboured on a conjugative IncA/C plasmid of ca. 120kb. This study confirmed that the resistance conferred by OXA-48 and VEB-8 contributed to the failure of antibiotic treatment and consequently death of the patient., (Copyright © 2015 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.)

We developed a two-step PCR-based strategy to detect genes encoding OqxAB, allowing a specific assignment of Tn6010-associated oqxAB in Enterobacteriaceae. Chromosomal location in this setup was confirmed by hybridization with I-CeuI-restricted genomes. This approach led us to find that Klebsiella sp. and Raoultella sp. reference strains chromosomally carried oqxAB., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

Hemolytic-uremic syndrome due to enterohemorrhagic Escherichia coli, belonging to serogroup O91 has rarely been described. We report here a case of post-diarrheal HUS due to EHEC O91 in an elderly patient for whom diagnosis was delayed given a previously diagnosed C. difficile infection. This case highlights the usefulness of Shiga-toxin detection., (Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The aim of this study was to investigate the prevalence of plasmid-mediated quinolone resistance (PMQR) determinants in Escherichia coli from neonatal intensive care units (NICUs) in Poland. The study was conducted on 80 E. coli isolates from different types of infections collected between 2009 and 2012. Six (5%) isolates were not susceptible to ciprofloxacin, 16% to ofloxacin, and 6.2% to levofloxacin. Among the 80 isolates, 27.5% carried at least one PMQR determinant (n = 22). qnrB was found in 8.8% of isolates (7/80), and qnrS was found in 2.5% (2/80). No isolates carrying qnrA, qnrD, qnrC, qepA, or oqxAB were found. The prevalence of aac(6')-Ib-cr was 27.5%, and 54.5% of the isolates contained aac(6')-Ib-cr and blaCTX-M. qnr-positive strains were more likely to produce extended-spectrum β-lactamases (ESBLs) than qnr-negative strains, which may suggest an association between qnr and ESBLs (88.9% vs. 17%, p = 0.0003). PMQR-positive isolates had significantly higher ciprofloxacin MIC50 values (28.8-fold higher when comparing the MIC50 values) than the PMQR-negative strains (0.23 vs. 0.008 mg/L), regardless of the presence of quinolone resistance-determining region mutations. Five of the nine isolates, for which conjugation was performed successfully, harbored plasmids that contained CTX-M together with qnr and aac(6')-Ib-cr. Our data suggest that the number of qnr genes detected in E. coli from newborns may be related to the selection of qnr through antimicrobial exposure. Even if fluoroquinolones are not commonly used in the NICU, E. coli isolates may carry PMQR. The high prevalence of PMQR is of serious concern, as it may be horizontally transferred to other pathogenic bacteria.

We present a patient with a necrotizing pneumonia due to Streptococcus pneumoniae. This complication often affects children and is relatively rare and unrecognized in adults. Its diagnosis is often difficult on chest-X-ray and need a chest computed tomography. No risk factors predisposing to necrosis are described in literature and its mortality is not different from pneumonia without necrosis. The serotype 3 is the most common type implicated in pneumococcal necrotizing pneumonia. Study of virulence factors of S. pneumoniae and various genetic polymorphisms of the host should allow a better understanding of this complication.

In this study, 922 consecutive non-duplicate clinical isolates of Enterobacteriaceae obtained from hospitalized and non-hospitalized patients at Bejaia, Algeria were analyzed for AmpC-type β-lactamases production. The ampC genes and their genetic environment were characterized using polymerase chain reaction (PCR) and sequencing. Plasmid incompatibility groups were determined by using PCR-based replicon typing. Phylogenetic grouping and multilocus sequence typing were determined for molecular typing of the plasmid-mediated AmpC (pAmpC) isolates. Of the isolates, 15 (1.6%) were identified as AmpC producers including 14 CMY-4-producing isolates and one DHA-1-producing Klebsiella pneumoniae. All AmpC-producing isolates co-expressed the broad-spectrum TEM-1 β-lactamase and three of them co-produced CTX-M and/or SHV-12 ESBL. Phylogenetic grouping and virulence genotyping of the E. coli isolates revealed that most of them belonged to groups D and B1. Multilocus sequence typing analysis of K. pneumoniae isolates identified four different sequence types (STs) with two new sequences: ST1617 and ST1618. Plasmid replicon typing indicates that blaCMY-4 gene was located on broad host range A/C plasmid, while LVPK replicon was associated with blaDHA-1. All isolates carrying blaCMY-4 displayed the transposon-like structures ISEcp1/ΔISEcp1-blaCMY-blc-sugE. Our study showed that CMY-4 was the main pAmpC in the Enterobacteriaceae isolates in Algeria., (Copyright © 2015 Elsevier Editora Ltda. All rights reserved.)

We characterized 53 OXA-48-producing Klebsiella pneumoniae (OXA-48-Kp) isolated between 2011 and 2013 in 21 French hospitals. All the isolates were genotyped using MLST and PFGE and the population structure of the species was determined by a nucleotide-based analysis of the entire K. pneumoniae MLST database. Most of the OXA-48-Kp isolates also produced CTX-M-15 and remained susceptible to imipenem and meropenem. The isolates were distributed into 20 STs, of which five were dominant (ST15, ST101, ST147, ST395 and ST405). All the OXA-48-Kp clustered in the major clade of K. pneumoniae KpI., (© 2014 The Authors Clinical Microbiology and Infection © 2014 European Society of Clinical Microbiology and Infectious Diseases.)

Ten carbapenem-resistant Enterobacteriaceae (eight Klebsiella pneumoniae isolates and two Enterobacter cloacae) isolates from Yemen were investigated using in vitro antimicrobial susceptibility testing, phenotypic carbapenemase detection, multilocus sequence typing (MLST) and replicon typing. Carbapenemase, extended-spectrum β-lactamase (ESBL) and plasmid-mediated quinolone resistance determinant genes were identified using PCR and sequencing. All of the 10 carbapenem-resistant Enterobacteriaceae were resistant to β-lactams, tobramycin, ciprofloxacin and cotrimoxazole. Imipenem, doripenem and meropenem MICs ranged from 2 to >32 mg l(-1) and ertapenem MICs ranged from 6 to >32 mg l(-1). All of the K. pneumoniae isolates showed ESBL activity in phenotypic tests. Genes encoding blaNDM were detected in all strains. All K. pneumoniae strains produced CTX-M-15 ESBL and SHV β-lactamases. TEM-1 β-lactamase was detected in seven isolates. Nine isolates were qnr positive including QnrB1, QnrA1 and QnrS1, and six isolates produced AAC-6'-Ib-cr. MLST identified five different sequence types (STs): ST1399, ST147, ST29, ST405 and ST340. Replicon typing showed the presence of IncFII1K plasmids in four transformants. To the best of our knowledge, this is the first report of NDM-1-producing Enterobacteriaceae isolates in Yemen., (© 2014 The Authors.)

The aim of this study was to describe the presence of plasmid-mediated quinolone resistance (PMQR) determinants in extended spectrum β-lactamase-producing Escherichia coli (ESBL-Ec) in hospitals in eastern France. All ESBL-Ec isolated from May 2008 to April 2009 in nine hospitals in eastern France were collected and screened by PCR for the presence of PMQR genes. bla genes were identified by PCR and sequencing. Randomly chosen PMQR-positive ESBL-Ec isolates were characterised by pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and phylogenetic group identification. Of the 447 ESBL-Ec tested, 122 (27.3%) had a PMQR determinant. aac(6')-Ib-cr was more prevalent (118/122; 96.7%) than qnr [4/122 (3.3%), comprising 2 qnrB2, 1 qnrB1 and 1 qnrA1]. Among 37 PMQR-positive ESBL-Ec isolates selected for typing, 26 (70%) carried bla CTX-M-15 of which 25/26 (96%) co-harboured aac(6')-Ib-cr. Of the 37 isolates, 14 belonged to the B2:ST131 clone, all of which produced AAC(6')-Ib-cr. On the other hand, most of the qnr genes (3/4) were observed in strains carrying bla genes other than bla CTX-M (qnrB2 and bla SHV-12 ; qnrA1 and bla TEM-52 ). These findings suggest that aac(6')-Ib-cr, rather than qnr, is the spreading PMQR determinant in ESBL-Ec isolates, and the qnr determinant is not specifically associated with bla CTX-M genes in France. aac(6')-Ib-cr was the predominant PMQR gene in ESBL-Ec isolated in eastern France hospitals, suggesting that the distribution may be due to clonal spread of ST131 CTX-M-15-producing E. coli isolates., (Copyright © 2014 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.)

qnrD is a plasmid mediated quinolone resistance gene from unknown origin, recently described in Enterobacteriaceae. It encodes a pentapeptide repeat protein 36-60% different from the other Qnr (A, B, C, S and VC). Since most qnrD-positive strains were described as strains belonging to Proteus or Providencia genera, we hypothesized that qnrD originated in Proteeae before disseminating to other enterobacterial species. We screened 317 strains of Proteeae for qnrD and its genetic support by PCR. For all the seven qnrD-positive strains (4 Proteus mirabilis, 1 Proteus vulgaris and 2 Providencia rettgeri) the gene was carried onto a small non-transmissible plasmid, contrarily to other qnr genes that are usually carried onto large multi-resistant plasmids. Nucleotide sequences of the qnrD-bearing plasmids were 96% identical. Plasmids contained 3 ORFs apart from qnrD and belonged to an undescribed incompatibility group. Only one plasmid, in P. vulgaris, was slightly different with a 1,568-bp insertion between qnrD and its promoter, leading to absence of quinolone resistance. We sought for similar plasmids in 15 reference strains of Proteeae, but which were tested negative for qnrD, and found a 48% identical plasmid (pVERM) in Providencia vermicola. In order to explain how qnrD could have been inserted into such native plasmid, we sought for gene mobilization structures. qnrD was found to be located within a mobile insertion cassette (mic) element which sequences are similar to one mic also found in pVERM. Our conclusions are that (i) the small non-transmissible qnrD-plasmids described here may result from the recombination between an as-yet-unknown progenitor of qnrD and pVERM, (ii) these plasmids are maintained in Proteeae being a qnrD reservoir (iii) the mic element may explain qnrD mobilization from non-transmissible plasmids to mobilizable or conjugative plasmids from other Enterobacteriaceae, (iv) they can recombined with larger multiresistant plasmids conjugated in Proteeae.

AAC(6')-Ib-cr is a plasmid-mediated quinolone resistance mechanism described worldwide for Escherichia coli. Since it confers in vitro only a low level of resistance to ciprofloxacin, we evaluated its impact on the in vivo activity of ciprofloxacin. Isogenic strains were obtained by transferring plasmid p449, harboring aac(6')-Ib-cr, into the quinolone-susceptible strain E. coli CFT073-RR and its D87G gyrA mutant. MICs were 0.015, 0.06, 0.25, and 0.5 μg/ml against E. coli strains CFT073-RR, CFT073-RR/p449, CFT073-RR GyrA(r), and CFT073-RR GyrA(r)/p449, respectively. Bactericidal activity was reduced at 1× the MIC for the three resistant derivatives, while at a fixed concentration of 0.5 μg/ml, 99.9% killing was observed for all strains except E. coli CFT073-RR GyrA(r)/p449. In the murine model of pyelonephritis, an optimal regimen of ciprofloxacin (10 mg/kg of body weight twice a day [b.i.d.]) significantly decreased the bacterial count in the kidneys of mice infected with E. coli CFT073 (1.6 versus 4.3 log10 CFU/g of kidney compared to untreated controls; P = 0.0001), while no significant decrease was observed for E. coli CFT073-RR/p449 (2.7 versus 3.1 log10 CFU/g; P = 0.84), E. coli CFT073-RR GyrA(r) (4.2 versus 4.1 log10 CFU/g; P = 0.35), or E. coli CFT073-RR GyrA(r)/p449 (2.9 versus 3.6 log10 CFU/g; P = 0.47). While pharmacokinetic and pharmacodynamic (PK/PD) parameters accounted for ciprofloxacin failure against gyrA-containing mutants, this was not the case for the aac(6')-Ib-cr-containing strains, suggesting an in situ hydrolysis of ciprofloxacin in the latter case.

We previously described a pyrosequencing-based method able to guarantee detection of aac(6')-Ib-cr by characterizing precisely the single nucleotide polymorphisms leading to Trp102Arg and Asp179Tyr. By gaining in simplicity and cost-effectiveness, through the use of real-time PCR, we propose here a cutting-edge evolution of our existing pyrosequencing method., (© 2013 Elsevier B.V. All rights reserved.)