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1. Evaluation of ceftolozane-tazobactam susceptibility on a French nationwide collection of Enterobacterales

Author

Jousset, Agnès B., primary, Bernabeu, Sandrine, additional, Emeraud, Cécile, additional, Bonnin, Rémy A., additional, Lomont, Alexandra, additional, Zahar, Jean Ralph, additional, Merens, Audrey, additional, Isnard, Christophe, additional, Soismier, Nathalie, additional, Farfour, Eric, additional, Fihman, Vincent, additional, Yin, Nicolas, additional, Barraud, Olivier, additional, Jacquier, Hervé, additional, Ranc, Anne-Gaëlle, additional, Laurent, Frédéric, additional, Corvec, Stéphane, additional, d'Epenoux, Louise Ruffier, additional, Bille, Emmanuelle, additional, Degand, Nicolas, additional, Plouzeau, Chloé, additional, Guillard, Thomas, additional, Cattoir, Vincent, additional, Mizrahi, Asaf, additional, Grillon, Antoine, additional, Janvier, Frédéric, additional, Brun, Cécile Le, additional, Amara, Marlène, additional, Bastide, Mathilda, additional, Lemonnier, Alban, additional, and Dortet, Laurent, additional

Published
2023
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2. Improvement of patient care through hirudotherapy and the management of leeches from their reception to their disposal in France

Author

Vaesken, Céline, primary, Besnier, Paul, additional, Bernardeau, Claire, additional, Garmi, Rachid, additional, Malherbe, Mélanie, additional, Mouet, Audrey, additional, Michon, Jocelyn, additional, Muzard, Alexandra, additional, Isnard, Christophe, additional, Bazin, Isaline, additional, and Saint-Lorant, Guillaume, additional

Published
2022
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4. Evolution of Bordetella pertussis over a 23-year period in France, 1996 to 2018

Author

Bouchez, Valérie, Guillot, Sophie, Landier, Annie, Armatys, Nathalie, Matczak, Soraya, Toubiana, Julie, Brisse, Sylvain, Brieu, Nathalie, Hamdad, Farida, Kempf, Marie, Pailhoriès, Hélène, Jensen, Cécile, Lehours, Philippe, Guiraud, Jennifer, Le Bars, Hervé, Isnard, Christophe, Wilhelm, Nathalie, Le Coustumier, Alain, Delmas, Julien, De Briel, Dominique, Souply, Laurent, Aberrane, Saïd, Coudé, Marie, Garnier, Fabien, Descours, Ghislaine, Jean-Pierre, Hélène, Alauzet, Corentine, Gibaud, Sophie-Anne, Bonacorsi, Stéphane, Brasme, Lucien, Lemée, Ludovic, Koebel, Christelle, Lanotte, Philippe, Bland, Stéphane, Petitprez, Hélène, Raffenot, Didier, Levast, Marion, Doucet-Populaire, Florence, Bourgeois-Nicolaos, Nadège, Burucoa, Christophe, Grattard, Florence, Marque-Juillet, Stéphanie, Biodiversité et Epidémiologie des Bactéries pathogènes - Biodiversity and Epidemiology of Bacterial Pathogens, Institut Pasteur [Paris], Centre national de Référence de la Coqueluche et autres Bordetelloses - National Reference Center for Whooping Cough and other Bordetella infections (CNR), Collège doctoral [Sorbonne universités], Sorbonne Université (SU), Département de Pédiatrie et maladies infectieuses [CHU Necker], CHU Necker - Enfants Malades [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Université de Paris (UP), This work was performed with the financial support of the Institut Pasteur and of Santé Publique France., French pertussis microbiology study group: Nathalie Brieu, Farida Hamdad, Marie Kempf, Hélène Pailhoriès, Cécile Jensen, Philippe Lehours, Jennifer Guiraud, Hervé Le Bars, Christophe Isnard, Nathalie Wilhelm, Alain Le Coustumier, Julien Delmas, Dominique De Briel, Laurent Souply, Saïd Aberrane, Marie Coudé, Fabien Garnier, Ghislaine Descours, Hélène Jean-Pierre, Corentine Alauzet, Sophie-Anne Gibaud, Stéphane Bonacorsi, Lucien Brasme, Ludovic Lemée, Christelle Koebel, Philippe Lanotte, Stéphane Bland, Hélène Petitprez, Didier Raffenot, Marion Levast, Florence Doucet-Populaire, Nadège Bourgeois-Nicolaos, Christophe Burucoa, Florence Grattard, Stéphanie Marque-Juillet, Institut Pasteur [Paris] (IP)-Université Paris Cité (UPCité), Collège Doctoral, Université Paris Cité (UPCité), and Institut Pasteur [Paris]-Institut Pasteur [Paris]

Subjects
Serotype, Bordetella pertussis, population evolution, Whooping Cough, Epidemiology, [SDV]Life Sciences [q-bio], Fimbria, MESH: Pertussis Toxin, MESH: Virulence Factors, Bordetella, Pertussis toxin, Microbiology, pertactin, MESH: Bordetella pertussis, 03 medical and health sciences, 0302 clinical medicine, Antigen, Virology, MESH: Child, medicine, Humans, Virulence Factors, Bordetella, 030212 general & internal medicine, Child, Whooping cough, 030304 developmental biology, Pertussis Vaccine, vaccine antigen deficiency, 0303 health sciences, Surveillance, MESH: Humans, biology, MESH: Bacterial Outer Membrane Proteins, Public Health, Environmental and Occupational Health, MESH: Whooping Cough, biology.organism_classification, medicine.disease, fimbriae serotype, 3. Good health, respiratory tract diseases, Vaccination, MESH: France, Pertussis Toxin, MESH: Pertussis Vaccine, France, Pertactin, Bacterial Outer Membrane Proteins
Abstract

Background Bordetella pertussis is the main agent of whooping cough. Vaccination with acellular pertussis vaccines has been largely implemented in high-income countries. These vaccines contain 1 to 5 antigens: pertussis toxin (PT), filamentous haemagglutinin (FHA), pertactin (PRN) and/or fimbrial proteins (FIM2 and FIM3). Monitoring the emergence of B. pertussis isolates that might partially escape vaccine-induced immunity is an essential component of public health strategies to control whooping cough. Aim We aimed to investigate temporal trends of fimbriae serotypes and vaccine antigen-expression in B. pertussis over a 23-year period in France (1996–2018). Methods Isolates (n = 2,280) were collected through hospital surveillance, capturing one third of hospitalised paediatric pertussis cases. We assayed PT, FHA and PRN production by Western blot (n = 1,428) and fimbriae production by serotyping (n = 1,058). Molecular events underlying antigen deficiency were investigated by genomic sequencing. Results The proportion of PRN-deficient B. pertussis isolates has increased steadily from 0% (0/38) in 2003 to 48.4% (31/64) in 2018 (chi-squared test for trend, p 481 insertion within the prn gene or a 22 kb genomic inversion involving the prn promoter sequence, indicative of convergent evolution. FIM2-expressing isolates have emerged since 2011 at the expense of FIM3. Conclusions B. pertussis is evolving through the rapid increase of PRN-deficient isolates and a recent shift from FIM3 to FIM2 expression. Excluding PRN, the loss of vaccine antigen expression by circulating B. pertussis isolates is epidemiologically insignificant.

Published
2021

5. Unexpected Cell Wall Alteration-Mediated Bactericidal Activity of the Antifungal Caspofungin against Vancomycin-Resistant Enterococcus faecium

Author

Isnard, Christophe, primary, Hernandez, Sara B., additional, Guérin, François, additional, Joalland, Fanny, additional, Goux, Didier, additional, Gravey, François, additional, Auzou, Michel, additional, Enot, David, additional, Meignen, Pierrick, additional, Giard, Jean-Christophe, additional, Cava, Felipe, additional, and Cattoir, Vincent, additional

Published
2020
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7. Enterococcus spp. : from opportunistic pathogens to probiotics

Author

Isnard, Christophe, STAR, ABES, Unité de Recherche Risques Microbiens (U2RM), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), Normandie Université, and Vincent Cattoir

Subjects
Oncobiotic, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Enterococcus hirae, Oncobiotique, Antibiotic resistance, Caspofungin, biochemical phenomena, metabolism, and nutrition, Immune response, [SDV.MP] Life Sciences [q-bio]/Microbiology and Parasitology
Abstract

Enterococci are commensal bacteria of the human and animal gastrointestinal tract, but some species as Enterococcus faecium, are also major opportunistic pathogens often multiply resistant to antibiotics. We studied the impact of non-antibiotic molecules widely used in intensive care units on fitness, virulence and resistance of a clinical isolate of E. faecium belonging to CC17 by a microscopic approach, a peptidoglycan analysis and a trancriptomic analysis. This work allowed us to demonstratethe antimicrobial effect of caspofungin, molecule known for its antifungal activity. We also characterized two novel resistance mechanisms found in E. faecium: i) resistance to lincosamides, streptogramines A and pleuromutilins (LSAP phenotype) linked to a point mutation in a gene encoding for a type-II ABC protein. ii) decreased susceptibility to tigecycline due to the occurrence of mutations within the rpsJ gene encoding the S10 ribosomal protein that plays a role in 30S ribosomal subunit formation. Finally, we participated to a study concerning Enterococcus hirae, species that induces the production of sub-populations of T lymphocytes that increase the in vivo efficacy of cyclophosphamide (CTX) in the treatment of murine tumors. A characterization of the virulence factors, antibiotic resistance profiles and colonization capacities of a collection of E. hirae isolates was carried out. A transcriptomic study in the presence of CTX and a comparative genomic study were also done, in order to characterize this species in view of its use as an oncobiotic., Les entérocoques, sont des bactéries commensales du tube digestif de l’homme et des animaux, mais certaines espèces, comme Enterococcus faecium, sont aussi des pathogènes opportunistes majeurs souvent multi-résistants aux antibiotiques. Nous avons étudié l’impact de molécules non antibiotiques utilisées dans les unités de soins intensifs, sur la virulence et la résistance d’une souche clinique de E. faecium par une approche microscopique, une analyse du peptidoglycane et une analyse trancriptomique. Ce travail nous a permis de décrire l’effet antibactérien de la caspofungine, molécule antifongique. Nous avons également étudié deux nouveaux mécanismes de résistance chez E. faecium i) la résistance aux lincosamides, streptogramines A et pleuromutilines (phénotype LSAP) par la mutation ponctuelle d’un gène codant pour une protéine ABC de type II. ii) la diminution de sensibilité à la tigécycline due à l’apparition de mutations au sein du gène rpsJ codant pour la protéine ribosomale S10 jouant un rôle dans la formation de la sous-unité 30S du ribosome. Enfin, nous avons participé à une étude sur Enterococcus hirae, espèce qui induirait la production de sous-populations de lymphocytes T permettant d’augmenter l’efficacité in vivo du cyclophosphamide (CTX) dans le traitement de tumeurs chez la souris. Une caractérisation des facteurs de virulence, de la résistance aux antibiotiques et du pouvoir de colonisation d’une collection de souches d’E. hirae a été menée, de même qu’une étude transcriptomique en présence de CTX et une étude de génomique comparative, afin de caractériser cette espèce dans l’optique de son utilisation comme oncobiotique.

Published
2017

9. Enterococcus hirae and Barnesiella intestinihominis Facilitate Cyclophosphamide-Induced Therapeutic Immunomodulatory Effects

Author

Daillère, Romain, primary, Vétizou, Marie, additional, Waldschmitt, Nadine, additional, Yamazaki, Takahiro, additional, Isnard, Christophe, additional, Poirier-Colame, Vichnou, additional, Duong, Connie P.M., additional, Flament, Caroline, additional, Lepage, Patricia, additional, Roberti, Maria Paula, additional, Routy, Bertrand, additional, Jacquelot, Nicolas, additional, Apetoh, Lionel, additional, Becharef, Sonia, additional, Rusakiewicz, Sylvie, additional, Langella, Philippe, additional, Sokol, Harry, additional, Kroemer, Guido, additional, Enot, David, additional, Roux, Antoine, additional, Eggermont, Alexander, additional, Tartour, Eric, additional, Johannes, Ludger, additional, Woerther, Paul-Louis, additional, Chachaty, Elisabeth, additional, Soria, Jean-Charles, additional, Golden, Encouse, additional, Formenti, Silvia, additional, Plebanski, Magdalena, additional, Madondo, Mutsa, additional, Rosenstiel, Philip, additional, Raoult, Didier, additional, Cattoir, Vincent, additional, Boneca, Ivo Gomperts, additional, Chamaillard, Mathias, additional, and Zitvogel, Laurence, additional

Published
2016
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13. Genetic characterization of a VanG-type vancomycin-resistant Enterococcus faecium clinical isolate.

Author

Sassi, Mohamed, Guérin, François, Lesec, Léonie, Isnard, Christophe, Fines-Guyon, Marguerite, Cattoir, Vincent, and Giard, Jean-Christophe

Subjects
ANTIBIOTICS, CHROMOSOMES, ERYTHROMYCIN, GENE expression, POLYMERASE chain reaction, RIFAMPIN, TETRACYCLINE, VANCOMYCIN, PHENOTYPES, VANCOMYCIN resistance, REVERSE transcriptase polymerase chain reaction, ENTEROCOCCUS faecium, SEQUENCE analysis, GENOTYPES
Abstract

Objectives: To characterize, phenotypically and genotypically, the first Enterococcus faecium clinical isolate harbouring a vanG operon.Methods: The antibiotic resistance profile of E. faecium 16-346 was determined and its whole genome sequenced using PacBio technology. Attempts to transfer vancomycin resistance by filter mating were performed and the inducibility of expression of the vanG operon was studied by reverse-transcription quantitative PCR (RT-qPCR) in the presence or absence of subinhibitory concentrations of vancomycin.Results: E. faecium 16-346 was resistant to rifampicin (MIC >4 mg/L), erythromycin (MIC >4 mg/L), tetracycline (MIC >16 mg/L) and vancomycin (MIC 8 mg/L), but susceptible to teicoplanin (MIC 0.5 mg/L). The strain harboured the vanG operon in its chromosome, integrated in a 45.5 kb putative mobile genetic element, similar to that of Enterococcus faecalis BM4518. We were unable to transfer vancomycin resistance from E. faecium 16-346 to E. faecium BM4107 and E. faecalis JH2-2. Lastly, transcription of the vanG gene was inducible by vancomycin.Conclusions: This is, to the best of our knowledge, the first report of a VanG-type vancomycin-resistant strain of E. faecium. Despite the alarm pulled because of the therapeutic problems caused by VRE, our work shows that new resistant loci can still be found in E. faecium. [ABSTRACT FROM AUTHOR]

Published
2018
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17. Genomic Analysis of Reduced Susceptibility to Tigecycline in Enterococcus faecium

Author

Cattoir, Vincent, Isnard, Christophe, Cosquer, Thibaud, Odhiambo, Arlène, Bucquet, Fiona, Guérin, François, and Giard, Jean-Christophe

Abstract

ABSTRACTTigecycline (TIG) is approved for use for the treatment of complicated intra-abdominal infections, skin and skin structure infections, as well as pneumonia. Acquired resistance or reduced susceptibility to TIG has been observed in Gram-negative rods, has seldom been reported in Gram-positive organisms, and has not yet been reported in Enterococcus faecium. Using the serial passage method, in vitromutant AusTig and in vitromutants HMtig1 and HMtig2 with decreased TIG susceptibility (MICs, 0.25 μg/ml) were obtained from strains E. faeciumAus0004 and HM1070 (MICs, 0.03 μg/ml), respectively. In addition, two vancomycin-resistant E. faeciumclinical isolates (EF16 and EF22) with reduced susceptibility to TIG (MICs, 0.5 and 0.25 μg/ml, respectively) were studied. Compared to the wild-type strains, the in vitromutants also showed an increase in the MICs of other tetracyclines. An efflux mechanism did not seem to be involved in the reduced TIG susceptibility, since the presence of efflux pump inhibitors (reserpine or pantoprazole) did not affect the MICs of TIG. Whole-genome sequencing of AusTig was carried out, and genomic comparison with the Aus0004 genome was performed. Four modifications leading to an amino acid substitution were found. These mutations affected the rpsJgene (efau004_00094, coding for the S10 protein of the 30S ribosomal subunit), efau004_01228(encoding a cation transporter), efau004_01636(coding for a hypothetical protein), and efau004_02455(encoding the l-lactate oxidase). The four other strains exhibiting reduced TIG susceptibility were screened for the candidate mutations. This analysis revealed that three of them showed an amino acid substitution in the same region of the RpsJ protein. In this study, we characterized for the first time genetic determinants linked to reduced TIG susceptibility in enterococci.

Published
2014
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18. Genetic Basis for In Vitroand In VivoResistance to Lincosamides, Streptogramins A, and Pleuromutilins (LSAP Phenotype) in Enterococcus faecium

Author

Isnard, Christophe, Malbruny, Brigitte, Leclercq, Roland, and Cattoir, Vincent

Abstract

ABSTRACTAs opposed to Enterococcus faecalis, which is intrinsically resistant to lincosamides, streptogramins A, and pleuromutilins (LSAP phenotype) by production of the ABC protein Lsa(A), Enterococcus faeciumis naturally susceptible. Since this phenotype may be selected for in vivoby quinupristin-dalfopristin (Q-D), the aim of this study was to investigate the molecular mechanism of acquired LSAP resistance in E. faecium. Six LSAP-resistant in vitromutants of E. faeciumHM1070 as well as three different pairs of clinical isolates (pre- and postexposure to Q-D) were studied. The full genome sequence of an in vitromutant (E. faeciumUCN90B) was determined by using 454 sequencing technology and was compared with that of the parental strain. Single-nucleotide replacement was carried out to confirm the role of this mutation. By comparative genomic analysis, a point mutation was found within a 1,503-bp gene coding for an ABC homologue showing 66% amino acid identity with Lsa(A). This mutation (C1349T) led to an amino acid substitution (Thr450Ile). An identical mutation was identified in all in vitroand in vivoresistant strains but was not present in susceptible strains. The wild-type allele was named eat(A) (for EnterococcusABC transporter), and its mutated allelic variant was named eat(A)v. The introduction of eat(A)vfrom UCN90B into HM1070 conferred the LSAP phenotype, whereas that of eat(A) from HM1070 into UCN90B restored susceptibility entirely. This is the first description of the molecular mechanism of acquired LSAP resistance in E. faecium. Characterization of the biochemical mechanism of resistance and the physiological role of this ABC protein need further investigations.

Published
2013
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19. Identification and Clinical Significance of Helcococcus kunziiin Human Samples

Author

Vergne, Anne, Guérin, François, Lienhard, Reto, Le Coustumier, Alain, Daurel, Claire, Isnard, Christophe, Marty, Nicole, Poyart, Claire, and Cattoir, Vincent

Abstract

ABSTRACTFrom 2008 to 2013, 39 Helcococcus kunziistrains were collected from human clinical specimens (79% from foot ulcers), and 85% of the 39 patients were infected. Matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry and molecular methods accurately identified all isolates. This large study of clinical observations confirms the potential pathogenic role of H. kunzii.

Published
2015
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20. In VitroActivity of Ceftolozane-Tazobactam against Enterobacter cloacaeComplex Clinical Isolates with Different β-Lactam Resistance Phenotypes

Author

Robin, Frédéric, Auzou, Michel, Bonnet, Richard, Lebreuilly, Romain, Isnard, Christophe, Cattoir, Vincent, and Guérin, François

Abstract

The study evaluated the in vitroactivity of ceftolozane-tazobactam (C/T) against 94 unique clinical isolates of Enterobacter cloacaecomplex (ECC). No difference was observed according to the ECC cluster.

Published
2018
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21. Antimicrobial Resistance in Enterobacterales Recovered from Urinary Tract Infections in France.

Author

Farfour E, Dortet L, Guillard T, Chatelain N, Poisson A, Mizrahi A, Fournier D, Bonnin RA, Degand N, Morand P, Janvier F, Fihman V, Corvec S, Broutin L, Le Brun C, Yin N, Héry-Arnaud G, Grillon A, Bille E, Jean-Pierre H, Amara M, Jaureguy F, Isnard C, Cattoir V, Diedrich T, Flevin E, Merens A, Jacquier H, Vasse M, and On Behalf Of The Gmc Study Group

Abstract

In the context of increasing antimicrobial resistance in Enterobacterales, the management of these UTIs has become challenging. We retrospectively assess the prevalence of antimicrobial resistance in Enterobacterales isolates recovered from urinary tract samples in France, between 1 September 2017, to 31 August 2018. Twenty-six French clinical laboratories provided the susceptibility of 134,162 Enterobacterales isolates to 17 antimicrobials. The most frequent species were E. coli (72.0%), Klebsiella pneumoniae (9.7%), Proteus mirabilis (5.8%), and Enterobacter cloacae complex (2.9%). The overall rate of ESBL-producing Enterobacterales was 6.7%, and ranged from 1.0% in P. mirabilis to 19.5% in K. pneumoniae , and from 3.1% in outpatients to 13.6% in long-term care facilities. Overall, 4.1%, 9.3% and 10.5% of the isolates were resistant to cefoxitin, temocillin and pivmecillinam. Cotrimoxazole was the less active compound with 23.4% resistance. Conversely, 4.4%, 12.9%, and 14.3% of the strains were resistant to fosfomycin, nitrofurantoin, and ciprofloxacin. However, less than 1% of E. coli was resistant to fosfomycin and nitrofurantoin. We identified several trends in antibiotics resistances among Enterobacterales isolates recovered from the urinary tract samples in France. Carbapenem-sparing drugs, such as temocillin, mecillinam, fosfomycin, cefoxitin, and nitrofurantoin, remained highly active, including towards ESBL-E.

Published
2022
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22. Genetic basis for in vitro and in vivo resistance to lincosamides, streptogramins A, and pleuromutilins (LSAP phenotype) in Enterococcus faecium.

Author

Isnard C, Malbruny B, Leclercq R, and Cattoir V

Subjects
ATP-Binding Cassette Transporters metabolism, Amino Acid Sequence, Bacterial Proteins metabolism, Base Sequence, Diterpenes pharmacology, Drug Resistance, Multiple, Bacterial drug effects, Enterococcus faecium drug effects, Enterococcus faecium metabolism, Genetic Complementation Test, Genotype, Molecular Sequence Data, Phenotype, Point Mutation, Polycyclic Compounds, Sequence Analysis, DNA, Sequence Homology, Amino Acid, Pleuromutilins, ATP-Binding Cassette Transporters genetics, Anti-Bacterial Agents pharmacology, Bacterial Proteins genetics, Drug Resistance, Multiple, Bacterial genetics, Enterococcus faecium genetics, Lincosamides pharmacology, Streptogramin A pharmacology
Abstract

As opposed to Enterococcus faecalis, which is intrinsically resistant to lincosamides, streptogramins A, and pleuromutilins (LSAP phenotype) by production of the ABC protein Lsa(A), Enterococcus faecium is naturally susceptible. Since this phenotype may be selected for in vivo by quinupristin-dalfopristin (Q-D), the aim of this study was to investigate the molecular mechanism of acquired LSAP resistance in E. faecium. Six LSAP-resistant in vitro mutants of E. faecium HM1070 as well as three different pairs of clinical isolates (pre- and postexposure to Q-D) were studied. The full genome sequence of an in vitro mutant (E. faecium UCN90B) was determined by using 454 sequencing technology and was compared with that of the parental strain. Single-nucleotide replacement was carried out to confirm the role of this mutation. By comparative genomic analysis, a point mutation was found within a 1,503-bp gene coding for an ABC homologue showing 66% amino acid identity with Lsa(A). This mutation (C1349T) led to an amino acid substitution (Thr450Ile). An identical mutation was identified in all in vitro and in vivo resistant strains but was not present in susceptible strains. The wild-type allele was named eat(A) (for Enterococcus ABC transporter), and its mutated allelic variant was named eat(A)v. The introduction of eat(A)v from UCN90B into HM1070 conferred the LSAP phenotype, whereas that of eat(A) from HM1070 into UCN90B restored susceptibility entirely. This is the first description of the molecular mechanism of acquired LSAP resistance in E. faecium. Characterization of the biochemical mechanism of resistance and the physiological role of this ABC protein need further investigations.

Published
2013
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