Marie Kempf

Emergence of resistance to carbapenems in Acinetobacter baumannii in Europe: clinical impact and therapeutic options

Despite having a reputation of low virulence, Acinetobacter baumannii is an emerging multidrug-resistant (MDR) pathogen responsible for community- and hospital-acquired infections that are difficult to control and treat. Interest in this pathogen emerged about one decade ago because of its natural MDR phenotype, its capability of acquiring new mechanisms of resistance and the existence of nosocomial outbreaks. Recent advances in molecular biology, including full genome sequencing of several A. baumannii isolates, has led to the discovery of the extraordinary plasticity of their genomes, which is linked to their great propensity to adapt to any environment, including hospitals. In this context, as well as the increasing antimicrobial resistance amongst A. baumannii isolates to the last-line antibiotics carbapenems and colistin, therapeutic options are very limited or absent in some cases of infections with pandrug-resistant bacteria. However, a large proportion of patients may be colonised by such MDR bacteria without any sign of infection, leading to a recurrent question for clinicians as to whether antibiotic treatment should be given and will be effective in the presence of resistance mechanisms. The worldwide emergence of A. baumannii strains resistant to colistin is worrying and the increasing use of colistin to treat infections caused by MDR bacteria will inevitably increase the recovery rate of colistin-resistant isolates in the future. Current knowledge about A. baumannii, including biological and epidemiological aspects as well as resistance to antibiotics and antibiotic therapy, are reviewed in this article, in addition to therapeutic recommendations.

Rapid Detection of Carbapenem Resistance in Acinetobacter baumannii Using Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry

Rapid detection of carbapenem-resistant Acinetobacter baumannii strains is critical and will benefit patient care by optimizing antibiotic therapies and preventing outbreaks. Herein we describe the development and successful application of a mass spectrometry profile generated by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) that utilized the imipenem antibiotic for the detection of carbapenem resistance in a large series of A. baumannii clinical isolates from France and Algeria. A total of 106 A. baumannii strains including 63 well-characterized carbapenemase-producing and 43 non-carbapenemase-producing strains, as well as 43 control strains (7 carbapenem-resistant and 36 carbapenem-sensitive strains) were studied. After an incubation of bacteria with imipenem for up to 4 h, the mixture was centrifuged and the supernatant analyzed by MALDI-TOF MS. The presence and absence of peaks representing imipenem and its natural metabolite was analyzed. The result was interpreted as positive for carbapenemase production if the specific peak for imipenem at 300.0 m/z disappeared during the incubation time and if the peak of the natural metabolite at 254.0 m/z increased as measured by the area under the curves leading to a ratio between the peak for imipenem and its metabolite being <0.5. This assay, which was applied to the large series of A. baumannii clinical isolates, showed a sensitivity of 100.0% and a specificity of 100.0%. Our study is the first to demonstrate that this quick and simple assay can be used as a routine tool as a point-of-care method for the identification of A. baumannii carbapenemase-producers in an effort to prevent outbreaks and the spread of uncontrollable superbugs.

Biotyping of multidrug-resistant Klebsiella pneumoniae clinical isolates from France and Algeria using MALDI-TOF MS.

Background Klebsiella pneumoniae is one of the most important pathogens responsible for nosocomial outbreaks worldwide. Epidemiological analyses are useful in determining the extent of an outbreak and in elucidating the sources and the spread of infections. The aim of this study was to investigate the epidemiological spread of K. pneumoniae strains using a MALDI-TOF MS approach. Methods Five hundred and thirty-five strains of K. pneumoniae were collected between January 2008 and March 2011 from hospitals in France and Algeria and were identified using MALDI-TOF. Antibiotic resistance patterns were investigated. Clinical and epidemiological data were recorded in an Excel file, including clustering obtained from the MSP dendrogram, and were analyzed using PASW Statistics software. Results Antibiotic susceptibility and phenotypic tests of the 535 isolates showed the presence of six resistance profiles distributed unequally between the two countries. The MSP dendrogram revealed five distinct clusters according to an arbitrary cut-off at the distance level of 500. Data mining analysis of the five clusters showed that K. pneumoniae strains isolated in Algerian hospitals were significantly associated with respiratory infections and the ESBL phenotype, whereas those from French hospitals were significantly associated with urinary tract infections and the wild-type phenotype. Conclusions MALDI-TOF was found to be a promising tool to identify and differentiate between K. pneumoniae strains according to their phenotypic properties and their epidemiological distribution. This is the first time that MALDI-TOF has been used as a rapid tool for typing K. pneumoniae clinical isolates.

Real-Time Sequencing To Decipher the Molecular Mechanism of Resistance of a Clinical Pan-Drug-Resistant Acinetobacter baumannii Isolate from Marseille, France

ABSTRACT We compare the whole-genome sequences of two multidrug-resistant clinical Acinetobacter baumannii isolates recovered in the same patient before (ABIsac_ColiS susceptible to colistin and rifampin only) and after (ABIsac_ColiR resistant to colistin and rifampin) treatment with colistin and rifampin. We decipher all the molecular mechanisms of antibiotic resistance, and we found mutations in the rpoB gene and in the PmrAB two-component system explaining resistance to rifampin and colistin in ABIsac_ColiR, respectively.

Carbapenem Resistance and Acinetobacter baumannii in Senegal: The Paradigm of a Common Phenomenon in Natural Reservoirs

Incidence of carbapenem-resistant Acinetobacter baumannii is rising in several parts of the world. In Africa, data concerning this species and its resistance to carbapenems are limited. The objective of the present study was to identify the presence of A. baumannii carbapenem-resistant encoding genes in natural reservoirs in Senegal, where antibiotic pressure is believed to be low. From October 2010 to January 2011, 354 human head lice, 717 human fecal samples and 118 animal fecal samples were screened for the presence of A. baumannii by real time PCR targeting bla OXA51-like gene. For all samples positive for A. baumannii, the carbapenemase-hydrolysing oxacillinases blaOXA23-like and blaOXA24-like were searched for and sequenced, and the isolates harbouring an oxacillinase were genotyped using PCR amplification and sequencing of recA gene. The presence of A. baumannii was detected in 4.0% of the head lice, in 5.4% of the human stool samples and in 5.1% of the animal stool samples tested. No bla OXA24 gene was detected but six fecal samples and three lice were positive for bla OXA23-like gene. The bla OXA23-like gene isolated in lice was likely a new oxacillinase sequence. Finally, the A. baumannii detected in stools were all of recA genotype 3 and those detected in lice, of recA genotype 4. This study shows for the first time a reservoir of bla OXA23-like-positive gene in human head lice and stool samples in Senegal.

Investigation of Acinetobacter baumannii resistance to carbapenems in Marseille hospitals, south of France: a transition from an epidemic to an endemic situation

Carbapenem‐resistant Acinetobacter baumannii infections are a worldwide endemic nosocomial threat. Between December 2010 and April 2011, an increase of carbapenem‐resistant A. baumannii infections occurred in several Marseille University Hospitals. The aim of this study was to investigate the increase of carbapenem‐resistant A. baumannii infections and to characterize the mechanisms of carbapenem resistance. The increase was detected by a homemade computer surveillance program, known as EPIMIC, that monitors antibiotic resistance profiles on a weekly basis. During this period, positive samples of carbapenem‐resistant A. baumannii were retrieved from patients hospitalized in different units. Genotyping of the isolates was performed using pulsed‐field gel electrophoresis (PFGE) and multi‐locus sequence typing (MLST), and carbapenemase gene analyses were performed to detect the presence of carbapenemases and to determine the relationships of the isolates. Carbapenem‐resistant A. baumannii were isolated in a total of 11 patients who were hospitalized in different hospitals units. We identified the presence of the blaOXA23‐like carbapenemase‐encoding gene in all of the isolates and found four major PFGE groups and different MLST groups. These results demonstrate a current evolution in the A. baumannii epidemiology in Marseille with a switch from an epidemic situation to an endemic situation and with several circulating clones.

Reservoirs of Non-baumannii Acinetobacter Species

Acinetobacter spp. are ubiquitous gram negative and non-fermenting coccobacilli that have the ability to occupy several ecological niches including environment, animals and human. Among the different species, Acinetobacter baumannii has evolved as global pathogen causing wide range of infection. Since the implementation of molecular techniques, the habitat and the role of non-baumannii Acinetobacter in human infection have been elucidated. In addition, several new species have been described. In the present review, we summarize the recent data about the natural reservoir of non-baumannii Acinetobacter including the novel species that have been described for the first time from environmental sources and reported during the last years.

Synergistic activity of sulbactam combined with colistin against colistin-resistant Acinetobacter baumannii

Athens ‘G. Gennimatas’, Athens, Greece); Sofia Tsiplakou, Vassiliki Papaioannou (Department of Microbiology, General Hospital of Attica ‘KAT’, Athens, Greece); Helen Fakiri, Nikoleta Skarmoutsou (Department of Microbiology, General Hospital of Athens ‘Sismanoglio’, Athens, Greece); Dimitris Kairis, Eleni Papoutsidou (Department of Microbiology, General Hospital of Attica ‘Asklipio Voulas’, Athens, Greece); Helen Katsifa, Fani Kesidou (Department of Microbiology, General Hospital ‘G. Papanikolaou’, Thessaloniki, Greece); Ekaterini Tsafaraki, Anna Tsouri (Department of Microbiology, General Hospital ‘Ag. Georgios’, Chania, Crete, Greece); Evangelia Platsouka, Zoi Roussou (Department of Microbiology, ‘Konstantopoulion’ General Hospital of Athens, Athens, Greece); and Helen Kaitsa-Tsiopoulou, Polizo Kazila (Department of Microbiology, ‘Theagenion’ Anticancer Hospital, Thessaloniki, Greece).

Detection of Acinetobacter baumannii in human head and body lice from Ethiopia and identification of new genotypes

BACKGROUNDnAcinetobacter baumannii has previously been detected and genotyped in human body lice. The objectives of this study were to determine the presence of this bacterium in head and body lice collected from healthy individuals in Ethiopia by molecular methods and to characterize the genotype.nnnMETHODSnHuman lice from locations at different altitudes in Ethiopia were screened for the presence of Acinetobacter sp by targeting the rpoB gene. Acinetobacter baumannii was detected and genotyped using recA PCR amplification.nnnRESULTSnA total of 115 head and 109 body lice were collected from 134 healthy individuals. Acinetobacter sp were found in 54 head (47%) and 77 body (71%) lice. The recA gene was sequenced for 60 of the Acinetobacter sp and 67% were positive for A. baumannii; genotype 1 was retrieved the most frequently.nnnCONCLUSIONnOur study is the first to show the presence of A. baumannii in human body lice, and also in head lice, in Ethiopia.