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We herein report the isolation and characterization of 21 Gram-stain-negative strains cultivated from the oropharynx of healthy children in Israel and Switzerland. Initially described as small colony variants of Kingella kingae, phenotypic analysis, biochemical analysis, phylogenetic analysis based on sequencing of the partial 16S rRNA gene and five housekeeping genes (abcZ, adk, G6PD, groEL and recA), and whole genome sequencing and comparison between members of the genera Kingella and Neisseria provided evidence for assigning them to the genus Kingella. Cellular fatty acids included important amounts of C12 : 0, C14 : 0, C16 : 0 and C16 : 1n7. Digital DNA-DNA hybridization between the isolates Sch538T and K. kingae ATCC 23330T revealed relatedness of 19.9 %. Comparative analysis of 16S rRNA gene sequences available in GenBank allowed matches to strains isolated in the USA, suggesting a wider geographical distribution. A novel species named Kingella negevensis sp. nov. is proposed, as most strains have been isolated in the Negev, a desert region of southern Israel. The type strain is Sch538T (=CCUG 69806T=CSUR P957).

Background: The aims of this study were to investigate Salmonella contamination in broiler chicken farms and slaughterhouses, to assess the antibiotic resistance profile in avian and human Salmonella isolates, and to evaluate the relationship between avian and human Extended Spectrum β-Lactamase (ESBL)-producing isolates. Salmonella was screened in different sample matrices collected at thirty-two chicken farms and five slaughterhouses. The human isolates were recovered from clinical specimens at the University Teaching Hospital of Constantine (UTH). All suspected colonies were confirmed by MALDI-TOF (Matrix Assisted Laser Desorption Ionization Time OF light) and serotyped. Susceptibility testing against 13 antibiotics including, amoxicillin/clavulanic acid, ticarcillin, cefoxitin, cefotaxime, aztreonam, imipenem, ertapenem, gentamicin, amikacin, ciprofloxacin, colistin, trimethoprim/sulfamethoxazole and fosfomycin, was performed using the disk diffusion method on Mueller-Hinton agar. ESBL-production was screened by the double-disk synergy test and confirmed by molecular characterization using PCR (polymerase chain reaction) amplification and sequencing of ESBL encoding genes. Clonality of the avian and human strains was performed using the Multi Locus Sequencing Typing method (MLST)., Results: Forty-five isolated avian Salmonella strains and 37 human collected ones were studied. Five S. enterica serotypes were found in avian isolates (mainly Kentucky) and 9 from human ones (essentially Infantis). 51.11% and 26.6% of the avian isolates were resistant to ciprofloxacin and cefotaxime, respectively, whereas human isolates were less resistant to these antibiotics (13.5% to ciprofloxacin and 16.2% to cefotaxime). Eighteen (12 avian and 6 human) strains were found to produce ESBLs, which were identified as bla CTX-M-1 (n = 12), bla CTX-M-15 (n = 5) and bla TEM group (n = 8). Interestingly, seven of the ESBL-producing strains (5 avian and 2 human) were of the same ST (ST15) and clustered together, suggesting a common origin., Conclusion: The results of the combined phenotypic and genotypic analysis found in this study suggest a close relationship between human and avian strains and support the hypothesis that poultry production may play a role in the spread of multidrug-resistant Salmonella in the human community within the study region.

We report the investigation methods for the diagnosis of an epidemic and culture-negative Kingella kingae endocarditis complicating a severe outbreak of hand, foot and mouth disease in a childcare center. The diagnosis was confirmed by polymerase chain reaction testing performed from cardiac tissue. Our findings argue for the systematic investigation of K. kingae outbreaks by using molecular tools in such context.

Purpose: The aim of the study was to investigate the prevalence and molecular support of carbapenem resistance in gram-negative bacilli clinical isolates collected between March 2013 and March 2015 in three cities (Annaba, Skikda, and Guelma) in northeastern Algeria., Results: One hundred eighty-six isolates were identified as Enterobacteriaceae (161), Pseudomonas aeruginosa (18), and Acinetobacter baumannii (7). Thirty-six of 186 (19.3%) were resistant to carbapenems. Among them, 11 harbored carbapenemase genes, including bla OXA-48 (2 Klebsiella pneumoniae), bla VIM-4 (2 P. aeruginosa), bla NDM-1 (2 A. baumannii), and bla OXA-23 (5 A. baumannii). In addition, other β-lactamases were detected: bla CTX-M-(15/66/139) , bla SHV-(28/85/1/133) , and bla TEM-1 . All imipenem-resistant P. aeruginosa displayed OprD mutations. Multilocus sequence typing demonstrated the presence of ST 404 and ST 219 in K. pneumoniae, ST 2 and ST 85 in A. baumannii, and ST (244, 1076, 241, 227, and 233) in P. aeruginosa., Conclusion: In this study, we report the first detection of P. aeruginosa ST 1076 harboring the bla VIM-4 gene in African countries in two cities (Annaba and Skikda) in northeastern Algeria. Additionally, we report the first detection of bla OXA-48 in K. pneumoniae ST 404 and ST 219 in Algerian cities (Annaba and Skikda).

Objectives: The present study aimed to screen for the presence of extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae in wild boars and Barbary macaques in Béjaïa and Jijel, Algeria., Methods: A total of 216 faecal samples collected between September 2014 and August 2015 were cultured on MacConkey agar supplemented with 1μg/mL ceftazidime. 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, and ESBLs were characterised by PCR and sequencing. Clonal relatedness was studied by multilocus sequence typing (MLST)., Results: A total of 47 ESBL-producing isolates were recovered from faecal samples from 40 (44%) of 90 wild boars and 7 (6%) of 126 from Barbary macaques, including 30 Escherichia coli and 17 Klebsiella pneumoniae. Results of PCR and sequencing analysis showed that all of the isolates produced CTX-M-15, and 25 isolates co-produced TEM-1. MLST demonstrated the presence of eight sequence types (STs) among the E. coli isolates (ST617, ST131, ST648, ST405, ST1431, ST1421, ST69 and ST226), whereas only one clone (ST584) was identified for all isolates of K. pneumoniae recovered from wild boars (n=10) and Barbary macaques (n=7)., Conclusions: This is the first report of CTX-M-15-producing E. coli and K. pneumoniae in wild animals from Algeria. The results show that African wildlife can act as a reservoir of the epidemic E. coli clone ST131 producing CTX-M-15, suggesting that this lineage can survive in different ecological niches and adapt to different hosts., (Copyright © 2016 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.)

Objectives: The aim of this study was to investigate the molecular epidemiology and the genetic support of carbapenem-resistant Enterobacteriaceae and Acinetobacter spp. isolates collected in the University Hospital of Ouargla, southern Algeria., Methods: A total of 99 Gram-negative bacteria (GNB) were collected from stool samples of colonised patients and from inanimate surfaces in the hospital environment between December 2014 and August 2015. Selected Enterobacteriaceae and Acinetobacter spp. isolates with reduced susceptibility to carbapenems were subjected to phenotypic study, including antibiotic susceptibility testing according to CA-SFM-EUCAST 2015 guidelines and modified Carba NP test. Genes encoding carbapenemases, extended-spectrum β-lactamases (ESBLs) and AmpC β-lactamases were screened by PCR and sequencing. Clonal relatedness was determined by multilocus sequencing typing (MLST)., Results: Of the 99 GNB isolates, 10 (10.1%) showed reduced susceptibility to carbapenems were studied further, including 7 Acinetobacter baumannii, 1 Acinetobacter nosocomialis, 1 Escherichia coli and 1 Klebsiella pneumoniae. PCR and sequencing showed that four A. baumannii isolates and the single A. nosocomialis isolate harboured bla NDM-1 . In addition, bla OXA-23 was observed in three A. baumannii isolates, and bla OXA-48 was detected in the two Enterobacteriaceae isolates. MLST assigned the K. pneumoniae to ST999 and the E. coli to ST38. The seven A. baumannii isolates belonged to ST85 and ST2., Conclusions: This study describes the epidemiology of carbapenemases produced by Enterobacteriaceae and Acinetobacter spp. in southern Algeria and reports the first description of metallo-β-lactamase NDM-1-producing A. nosocomialis in Algeria., (Copyright © 2016 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.)

The emergence and the global spread of carbapenemases concern to health services worldwide. Their celestial rise among Gram-negative bacilli has challenged both the scientific and pharmaceutical sectors. Indeed, infections caused by these bacteria have limited treatment options and have been associated with high mortality and morbidity rates. Carbapenemase producers are mainly identified among Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, and Acinetobacter baumannii and still mostly in hospital settings and rarely in the community. They are closely related to KPC, VIM, IMP, NDM, and OXA-48 types. The encoding genes are mostly plasmid located and associated with various mobile genetic elements. The Mediterranean area is of interest due to a great diversity and population mixing. The prevalence of carbapenemases is particularly high and variant among countries, partially depending on the population exchange relationship between the regions and the possible reservoirs of each carbapenemase. This review described the epidemiology of carbapenemases in this region of the world highlighting the worrisome situation and the need to screen and detect these enzymes to prevent and control their dissemination especially as it is clear that very few novel antibiotics will be introduced in the next few years, making the dissemination of carbapenem-resistant Gram-negative bacteria of crucial importance worldwide.

This study was designed to investigate environmental colonisation in Algerian hospitals by carbapenem-resistant Gram-negative bacilli (GNB), including molecular characterisation of their resistance, and to perform a comparative molecular analysis between clinical and environmental strains. GNB isolated from hospitalised patients and the hospital environment were identified using microbiological methods and matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF/MS). Antibiotic susceptibility testing was performed by disk diffusion and Etest methods. Carbapenemase- and extended-spectrum β-lactamase (ESBL)-encoding genes were searched for using PCR and sequencing. Clonality of the environmental and clinical strains was assessed by multilocus sequencing typing (MLST). A total of 32 carbapenem-resistant GNB were isolated, including 16 (29%) of 56 multidrug-resistant (MDR) GNB from clinical specimens and 16 (48%) of 33 MDR-GNB from inanimate surfaces. Of the 32 carbapenem-resistant isolates, 14 produced a carbapenemase. The bla OXA-48 gene was detected both in clinical and surface isolates of Klebsiella pneumoniae (n=3) and Enterobacter cloacae (n=2). Clinical and surface isolates of Acinetobacter baumannii were found to produce the carbapenemases NDM-1 (7 isolates) and OXA-23 (2 isolates). MLST revealed clonal diversity and a relationship between environmental and clinical strains with identical sequence types. Here we report the first description of an OXA-48-producing E. cloacae isolate in Algeria. We also highlight the important role of inanimate surfaces in the spread of carbapenem-resistant bacteria and the emergence of nosocomial infections., (Copyright © 2016 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.)

The aim of this study was to characterise the mechanisms of carbapenem resistance in Acinetobacter baumannii strains isolated in an Algerian hospital. A total of 43 imipenem-resistant A. baumannii clinical isolates collected between 2010 and 2013 were identified using API 20NE and were confirmed by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF/MS). Antibiotic susceptibility testing was performed by the disk diffusion and Etest methods. Carbapenemase activity was detected using microbiological tests and PCR. Genetic transfer of the blaNDM-1 gene was performed by conjugation using sodium azide-resistant Escherichia coli J53 as recipient strain. Clonal relationships were studied by multilocus sequence typing (MLST) using partial sequences of the csuE and blaOXA-51 genes. All 43 A. baumannii isolates were resistant to imipenem with high minimum inhibitory concentrations (MICs) (>32μg/mL). The strains harboured blaOXA-23, blaNDM-1, blaOXA-58 and/or blaOXA-24 genes. Co-existence of blaNDM-1 and blaOXA-23 or blaOXA-58 was detected in two isolates and one isolate, respectively. NDM-1 plasmid transfer to E. coli J53 was successful only for one of the three strains harbouring both blaNDM-1 and blaOXA-23 or blaOXA-58. The phylogenetic tree obtained from concatenation of the partial sequences of csuE and blaOXA-51 showed that there was no genetic relationship between the isolates and the blaNDM-1 resistance gene. Here we report for the first time the co-occurrence of blaNDM-1 along with blaOXA-23 or blaOXA-58 in recent clinical isolates of A. baumannii from Northeast Algeria. These findings re-emphasise the dissemination and rapid spread of blaNDM-1 carbapenemase genes in multidrug-resistant clinical A. baumannii isolates in Algeria., (Copyright © 2016 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.)

This study aimed to characterise the molecular support of antibiotic resistance in expanded-spectrum cephalosporin (ESC)-resistant Escherichia coli isolates recovered from healthy broilers in Béjaïa, northeast Algeria. A total of 61 intestinal swabs from slaughtered broilers from four regions in Béjaïa locality, Algeria, were collected between February and April 2014, from which 20 ESC-resistant E. coli strains were isolated. Escherichia coli isolates were identified by classical biochemical and MALDI-TOF methods. Antibiotic susceptibility testing was performed using disk diffusion and Etest methods. Screening for β-lactamases, aminoglycoside-modifying enzyme (AME)-encoding genes and qnr determinants was performed by PCR and sequencing. Clonal relatedness was determined using molecular typing by multilocus sequence typing (MLST). Antibiotic susceptibility testing revealed that the isolates showed high rates of resistance (>90%) to amoxicillin, amoxicillin/clavulanic acid, piperacillin/tazobactam, aztreonam, ceftazidime, streptomycin, tobramycin, nalidixic acid and ciprofloxacin. Low rates of resistance were observed for kanamycin (35%), amikacin (30%), cefoxitin (20%) and cefotaxime (15%). Molecular characterisation revealed that all of the isolates expressed the blaTEM-1 gene. Fourteen of them harboured the blaSHV-12 gene, two harboured the blaCTX-M-1 gene and four isolates harboured blaCMY-2. Screening for AME-encoding genes demonstrated that all isolates contained the aadA gene. In addition, qnrA was detected as the quinolone resistance determinant in 13 isolates. MLST revealed four known sequence types (STs), including ST744, ST38, ST1011 and ST2179, as well as one new sequence type (ST5086). Here we report the first study describing the clonal diversity of extended-spectrum β-lactamase (ESBL)- and plasmid AmpC-producing E. coli isolated from healthy broilers in Algeria., (Copyright © 2016 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.)

Introduction: The aim of the study was to investigate the prevalence of extended-spectrum β-lactamase (ESBL) and carbapenemase production among clinical isolates of Enterobacteriaceae recovered from Tunisian and Libyan hospitals., Methodology: Bacterial isolates were recovered from patients in intensive care units and identified by biochemical tests and MALDI-TOF. Antibiotic susceptibility testing was performed by disk diffusion and the E-test method. ESBL and carbapenemase activities were detected using standard microbiological tests. Antibiotic resistance-encoding genes were screened by PCR and sequencing. Clonal relationships between Klebsiella pneumoniae strains were carried out using multi-locus sequence typing (MLST)., Results: A total of 87 isolates were characterized, with 51 and 36, respectively, identified as E. coli and K. pneumoniae. Overall the resistance prevalence was high for aminoglycosides (> 60%), fluoroquinolones (> 80%), and extended-spectrum cephalosporins (> 94%), and was low for imipenem (11.4%). Among this collection, 58 strains (66.6%) were ESBL producers and 10 K. pneumoniae strains (11.4%) were carbapenemase producers. The antibiotic resistance-encoding genes detected were blaCTX-M-15 (51.7%), blaTEM-1 (35.6%), several variants of blaSHV (21.8%), and blaOXA-48 (11.4%). The MLST typing of K. pneumoniae isolates revealed the presence of multiple clones and three novel sequence types. Also, close relationships between the OXA-48-producing strains from Tunisia and Libya were demonstrated., Conclusions: This study is the first paper describing the emergence of carbapenemase- and ESBL-producing Enterobacteriaceae, sensitive to colistin, isolated in Tunisia and Libya. Active surveillance and testing for susceptibility to colistin should be implementing because resistance to colistin, mainly in Klebsiella, has been recently reported worldwide.

Acinetobacter baumannii is an opportunistic pathogen causing various nosocomial infections. The aim of this study was to characterise the molecular support of carbapenem-resistant A. baumannii clinical isolates recovered from two Libyan hospitals. Bacterial isolates were identified by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF/MS). Antibiotic susceptibility testing was performed using disk diffusion and Etest methods, and carbapenem resistance determinants were studied by PCR amplification and sequencing. Multilocus sequence typing (MLST) was performed for typing of the isolates. All 36 imipenem-resistant isolates tested were identified as A. baumannii. The blaOXA-23 gene was detected in 29 strains (80.6%). The metallo-β-lactamase blaNDM-1 gene was detected in eight isolates (22.2%), showing dissemination of multidrug-resistant (MDR) A. baumannii in Tripoli Medical Center and Burn and Plastic Surgery Hospital in Libya, including one isolate that co-expressed the blaOXA-23 gene. MLST revealed several sequence types (STs). Imipenem-resistant A. baumannii ST2 was the predominant clone (16/36; 44.4%). This study shows that NDM-1 and OXA-23 contribute to antibiotic resistance in Libyan hospitals and represents the first incidence of the association of these two carbapenemases in an autochthonous MDR A. baumannii isolated from patients in Libya, indicating that there is a longstanding infection control problem in these hospitals., (Copyright © 2016 Elsevier B.V. and International Society of Chemotherapy. All rights reserved.)

Actinobaculum massiliense (Euzéby, 2006) was isolated from the urine of an elderly woman in 2001. Unfortunately, the strain deposited as the type strain was, by error, an Actinobaculum schaalii strain (Yassin et al., 2015). In 2015, we isolated a new strain of A. massiliense, FC3, from the urine of a 12-year-old patient with acute cystitis. We herein present the characteristics of strain FC3 (=CSUR P1982=DSM 100580) and formally propose it as the neotype strain of A. massiliense.

Strain FF10(T) (= CSUR P1489 = DSM 100884) was isolated from the oral cavity of a lizard (Varanus niloticus) in Dakar, Senegal. Here we used a polyphasic study including phenotypic and genomic analyses to describe the strain FF10(T). Results support strain FF10(T) being a Gram-positive coccus, facultative anaerobic bacterium, catalase-negative, non-motile and non-spore forming. The sequenced genome counts 2.46 Mb with one chromosome but no plasmid. It exhibits a G+C content of 40.4% and contains 2471 protein-coding and 45 RNA genes. On the basis of these data, we propose the creation of Streptococcus varani sp. nov.

Uterine leiomyomas rarely present as gynaecological emergencies. We report a case of a 29-year-old nulliparous woman, with a negative pregnancy test, who presented with collapse and an admission haemoglobin count of 68 g/L. Urgent CT of the abdomen revealed a 14 × 19 × 10 cm uterine fibroid and intraperitoneal free fluid. Emergency laparotomy confirmed massive intraperitoneal haemorrhage from a large serosal vessel on top of a 19 cm subserosal fibroid. A myomectomy was performed, preserving this patient's fertility. Histopathology confirmed a benign leiomyoma with hydropic changes., (2016 BMJ Publishing Group Ltd.)

Actinobaculum massiliense strain FC3 was isolated from the urine of a patient with acute cystitis. The 2.06-Mb genome of strain FC3 contains 17 toxin/antitoxin modules and 9 bacteriocin-encoding genes that may play a role in virulence. The genome also exhibits 693 genes acquired by lateral gene transfer., (Copyright © 2016 Beye et al.)

In recent years, the number of multidrug-resistant bacteria has increased rapidly and several epidemics were signaled in different regions of the world. Faced with this situation that presents a major global public health concern, the development and the use of new and rapid technologies is more than urgent. The use of the next-generation sequencing platforms by microbiologists and infectious disease specialists has allowed great progress in the medical field. Here, we review the usefulness of whole-genome sequencing for the detection of virulence and antibiotic resistance associated genes.

The aim of the present study was to investigate the molecular mechanism of carbapenem resistance in Pseudomonas aeruginosa and Acinetobacter baumannii clinical isolates recovered from Libyan hospitals between April 2013 and April 2014. In total, 49 strains (24 P. aeruginosa and 25 A. baumannii) were isolated, including 21 P. aeruginosa and 22 A. baumannii isolates (87.75%) resistant to imipenem (minimum inhibitory concentrations ≥16 μg/ml). The blaVIM-2 gene was detected in 19 P. aeruginosa isolates. All imipenem-resistant P. aeruginosa isolates showed the presence of OprD mutations. Acquired OXA-carbapenemase-encoding genes were present in all A. baumannii isolates: blaOXA-23 (n=19) and blaOXA-24 (n=3). Finally, a total of 13 and 17 different sequence types were assigned to the 21 P. aeruginosa and the 22 A. baumannii carbapenem-resistant isolates, respectively. This study is the first report describing imipenem-resistant P. aeruginosa and A. baumannii isolated from patients in Libya. We report the first case of co-occurrence of blaVIM-2 with oprD porin loss in identical isolates of P. aeruginosa in Libya and demonstrate that these oprD mutations can be used as a tool to study the clonality in P. aeruginosa isolates. We also report the first identification of multidrug-resistant A. baumannii isolates harboring blaOXA-23-like, blaOXA-24-like, and blaOXA-48-like genes in Libya.

Aim: The aim of this study was to investigate the prevalence and the mechanisms of carbapenem and colistin resistance in Acinetobacter baumannii clinical isolates in an Algerian hospital., Results: Twelve isolates were collected between October 2013 and March 2014. All isolates were resistant to almost all antibiotics tested with a high-level resistance to imipenem (minimum inhibitory concentrations [MICs] >32 mg/L) with one strain showing resistance to colistin (MIC=16 mg/L). The results of the modified Hodge test and the modified Carba NP test were positive for all isolates. Besides, the activity of β-lactamases was inhibited by EDTA in only two isolates. All the 12 isolates contained the naturally occurring blaOXA-51-like gene. Ten of them harbored the OXA β-lactamases: blaOXA-23 (six isolates) and blaOXA-24 (four isolates) genes, while two isolates were positive for blaNDM-1 gene. The colistin-resistant isolate producing OXA-24 enzyme harbored a single mutation in the pmrB gene. Multilocus sequence typing demonstrated that the 12 isolates belonged to 2 clones: 10 to ST2 and 2 to ST85., Conclusions: Here, we describe the mechanisms of carbapenem resistance and we report the first colistin and carbapenemase-producing A. baumannii clinical isolate from a patient in Algeria.

The main objective of this study was to decipher the molecular mechanism of resistance to carbapenems and aminoglycosides in a large series of 150 Acinetobacter baumannii clinical isolates collected from July 2012 to September 2013 in Egypt. We report for the first time the emergence of bla(NDM-1) and the cooccurrence of 16S rRNA methylase armA with bla(NDM-1) and bla(OXA-23) in Egyptian hospitals. Multilocus sequence typing identified 27 distinct sequence types, 11 of which were novel., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

The emergence of colistin-resistant Klebsiella pneumoniae (CRKP) is a major public health concern worldwide. In this study, the prevalence and molecular basis of colistin resistance in CRKP isolated from healthy individuals and patients in Lao PDR, Thailand, Nigeria and France were investigated. Stool samples were screened by culture for the presence of colistin-resistant Klebsiella spp. Whole-genome sequence analysis was used to decipher the molecular mechanism of colistin resistance in a blaNDM-1-positive in vitro-selected CRKP mutant. PCR amplification and sequencing of the mgrB genetic environment was performed for all CRKP isolates as well as control colistin-susceptible K. pneumoniae (CSKP) isolates recovered from the same stools. A total of 869 stool samples were screened for colistin-resistant Klebsiella spp., yielding 32 CRKP and 2 colistin-resistant Klebsiella oxytoca. Comparative whole-genome sequence analysis revealed that an in vitro-selected CRKP mutant had an insertion sequence in its mgrB gene, as well as missense mutations in other selected clones. Of the 34 colistin-resistant Klebsiella spp. isolates, 14 (41.2%; 13 CRKP and 1 K. oxytoca) from the four countries also had various defects in their mgrB genes, but no such defects were found in the CSKP controls (P<10(-4)). Few mutations were observed in pmrAB compared with mgrB among the CRKP isolates. The worldwide emergence of CRKP is a major public health concern. Detection and surveillance of such strains are warranted to prevent an uncontrollable pandemic. Inactivation of the PhoP/PhoQ regulator gene mgrB is associated with ≥40% of colistin resistance among the CRKP isolates observed in this study., (Copyright © 2014 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.)

Purpose: The aim of this study was to investigate the molecular support of resistance to carbapenems, aminoglycosides, and fluoroquinolones in Acinetobacter baumannii clinical isolates collected from Yemen hospital., Methods: Three A. baumannii were isolated in February 2013 from three patients hospitalized at Al-Thawra University Hospital in Sana'a, Yemen. Antibiotic susceptibility testing was performed using the disk diffusion and E-test methods. Carbapenemase production was carried out by the modified Hodge test (MHT) and imipenem-ethylenediaminetetraacetic acid (EDTA) methods. Carbapenem, aminoglycoside, and fluoroquinolone resistance determinants were studied by polymerase chain reaction and sequencing. The epidemiological relatedness of the three strains was studied using multilocus sequence typing (MLST)., Results: The isolates were resistant to almost all antibiotics tested with very high imipenem, amikacin, and ciprofloxacin minimum inhibitory concentrations (>32, >256, and >32 mg/L, respectively). The microbiological tests showed that the three A. baumannii were MHT positive, besides, the activity of β-lactamases was not inhibited by EDTA. All the three isolates contained the naturally occurring bla(OXA-51)-like gene and the bla(OXA-23)-like carbapenemase-encoding gene. The 16S rRNA methylase armA gene was detected in the three isolates. In addition, screening for genes encoding the aminoglycoside-modifying enzymes (AMEs) demonstrated that one isolate contained the acetyltransferase gene aac(6')-Ib. Fluoroquinolone resistance was associated with a single mutation Ser83Leu in the quinolone resistance determining region of the gyrA gene in all isolates. The MLST showed that the sequence type (ST) obtained corresponds to ST2 for the three strains., Conclusions: Here we report the first identification of multidrug-resistant A. baumannii isolates harboring the bla(OXA-23)-like gene, AMEs [aac(6')-Ib], and the 16S rRNA methylase (armA) in the Yemen hospital.

We report class A carbapenemase (KPC)-3-producing Klebsiella pneumoniae meningitis in a 6-month-old child in Algeria. Multilocus sequence typing showed that the sequence type obtained corresponded to ST512, an allelic single-locus variant of the pandemic ST258 widely distributed in KPC producers from Europe. To our knowledge, this is the first report of KPC-3-producing K. pneumoniae ST512 in a North African country.

Purpose: The aim of the present study was to characterize the molecular support of resistance to carbapenems, aminoglycosides and fluoroquinolones in carbapenem-resistant Acinetobacter baumannii clinical isolates recovered between January 2011 and April 2013 from Algerian hospitals., Methods: Antibiotic susceptibility testing was performed using disk diffusion and Etest methods. Carbapenemase activity was detected using both MALDI-TOF mass spectrometry assay and via microbiological tests. Carbapenem, aminoglycoside and fluoroquinolone resistance determinants were studied by PCR and sequencing. Clonal relationships between strains were determined using Multi Locus Sequence Typing (MLST)., Results: A total of 47 imipenem-resistant A. baumannii were isolated and identified by MALDI-TOF mass spectrometry. All imipenem-resistant strains were positive in the modified Hodge test, and EDTA inhibited the activity of metallo-β-lactamases enzymes in 11 strains. The blaOXA-23 gene was detected in 33 strains and the blaOXA-24 gene in 10 strains. The metallo-β-lactamase blaNDM-1 gene was detected in 11 isolates (23.4%) from Algiers and Sétif, including 7 that co-expressed a blaOXA-23 gene. Resistance to aminoglycosides was due to the production of aminoglycoside-modifying enzymes, AAC(3)-Ia, AADA, ANT(2″)-I, APH(3')-VI, and 16S rRNA methylases, ArmA. The fluoroquinolone resistance was mainly associated with mutations at Ser83Leu and Ser80Leu of the gyrA and parC genes, respectively. MLST revealed five sequence types (STs), 1, 2, 19, 25, and 85. The imipenem-resistant A. baumannii ST2 was the predominant clone (35/47)., Conclusions: Here we report for the first time clinical multidrug-resistant A. baumannii isolates harboring 16S rRNA methylase gene, armA, and rapid spread of metallo-β-lactamase NDM-1 isolated from patients in Algeria., (Copyright © 2014. Published by Elsevier Ltd.)

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.)

Antibiotic susceptibility testing was performed on 71 Acinetobacter baumannii clinical isolates, and presence of antibiotic resistance genes was screened for by PCR amplification and sequencing. Resistance rates were very high for aminoglycosides (22-80%), fluoroquinolones (>90%), and cephalosporins (>90%) but remained low for rifampin (2.8%) or null for colistin. Antibiotic resistance encoding genes detected were as follows: blaTEM-128 gene (74.6%), aph(3')-VI (50.7 %), aadA (63.4%), ant(2″)-I (14.1%), aac(3)-Ia (91.1%), aac(6')-Ib (4.2%), mutation Ser83Leu in gyrA (94.4%), double mutations Ser83Leu and Ser80Leu (or Ser84Leu) in gyrA and parC (69.0%), and mutation I581N in RRDR of the rpoB gene., (Copyright © 2013 Elsevier Inc. All rights reserved.)

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.

Background: Chuvash-type polycythemia is a rare blood disease associated with an increased tendency toward thrombosis and its resulting problems, which at times can be life threatening. The main goal of treatment is to prevent thrombosis and minimize the complications of therapy., Case: A 20-year-old, Asian woman with her first pregnancy was admitted to the hospital at 16 weeks' gestation. Her initial blood work showed a hemoglobin level of 19 g/dL, hematocrit of 55% and erythropoietin level of 21.9 IU/L (laboratory reference range, 5-25). The results of a molecular analysis of the blood specimen confirmed the diagnosis of Chuvash-type polycythemia due to von Hippel-Lindau gene mutation. Weekly venisection until delivery was planned. Subsequently, the hemoglobin dropped to 15 g/dL and hematocrit to 44.6%. A 30-week scan demonstrated static growth below the third centile and oligohydramnios but normal Doppler findings. Prophylactic low-molecular-weight heparin was started. The next growth scan, at 32 weeks, showed an improvement in abdominal circumference. Three days later, fetal heart monitoring showed an abnormal pattern requiring an emergency cesarean section., Conclusion: Early institution of heparin is beneficial in the management of Chuvash-type polycythemia in pregnancy as the condition is thrombogenic.

Background: A rectal tear with an intact anal sphincter is an unusual presentation. Failure to recognize or repair perineal and rectal lesions increases the chance of fistulas and incontinence., Case: A 37-year-old woman, para 1, was admitted with spontaneous onset of labor at 36 weeks following an uncomplicated pregnancy. During the active second stage of labor, spurting of amniotic fluid through the anus was noted. Labor progressed normally, and the patient delivered a healthy infant of 2.5 kg. Immediate per rectal examination revealed a large rectal tear with an undamaged anal sphincter. The defect was repaired, and the patient recovered completely., Conclusion: Systematic examination of the perineum, vagina and rectum should be done to assess the severity of damage after all vaginal deliveries in order to recognize occult as well as visible damage. It should be treated promptly to reduce postpartum morbidity.

In this article we present the protocol of the Birmingham Registry for Twin Heritability Studies (BiRTHS), which aims to establish a long-term prospective twin registry with twins identified from the antenatal period and subjected to detailed follow-up. We plan to investigate the concordance in anthropometrics and early childhood phenotypes between 66 monozygotic and 154 dizygotic twin pairs in the first 2 years of recruitment. In this project we plan to determine the relative contributions of heritability and environment to fetal growth, birth size, growth in infancy and development up to 2 years of age in an ethnically mixed population. Twins will be assessed with the Griffitth's Mental Development Scales, which will enable us to obtain detailed information on development. As maternal depression may have an effect on the twins' neurodevelopment, the Edinburgh Postnatal Depression Scale will be used at various stages during pregnancy and after delivery to assess maternal depressive symptoms. The increasing prevalence of obesity in both adults and children has raised concerns about the effect of maternal obesity in pregnancy on fetal growth. The prospective study design gives us the opportunity to obtain data on maternal nutrition (reflected by body mass index) and ante- and postnatal growth and development of twins.

Objectives: To evaluate the significance of various risk factors associated with endometrial polyps in women with abnormal uterine bleeding., Methods: A prospective cohort study of 248 consecutive patients evaluated for abnormal uterine bleeding in a Rapid Access Ambulatory Diagnostic (RAAD) clinic was carried out from 1996 to 1997. Endometrial polyps were diagnosed using outpatient hysteroscopy and their histological nature was confirmed in all patients by performing inpatient polypectomy. Multivariate logistic regression modeling was used to evaluate the effects of age, parity, menopausal status, hormone replacement therapy and tamoxifen treatment on the occurrence of polyps., Results: In an analysis adjusted for the effects of age, parity and menopausal status, tamoxifen treatment was associated with endometrial polyps (adjusted odds ratio 11.21, 95% confidence interval 2.70-46.46, P=0.0009) but hormone replacement therapy was not (adjusted odds ratio 1.48, 95% confidence interval 0.68-3.20, P=0.32)., Conclusion: Our study confirmed that tamoxifen is associated with endometrial polyps. However, it rejects the hypothesis that hormone replacement therapy is a risk factor for endometrial polyps.

We examined the relative significance of hysteroscopic and ultrasonographic evidence of endometrial atrophy in relation to insufficient sample on outpatient endometrial biopsy in women with abnormal uterine bleeding. Multivariate logistic regression modelling was used to evaluate the independent effects of age, menopausal status, hysteroscopic findings and sonographic endometrial thickness on outpatient endometrial sampling (sufficient or insufficient) used as the binary dependent variable. Insufficient sample on endometrial biopsy was associated with hysteroscopic finding of endometrial atrophy (OR 4.79, 95% CI 1.05-21.91, P = 0.04) and sonographic endometrial thickness below 5 mm (OR 0- 19. 95% CI 0.07-0.53, P = 0.001). There was no association with patient's age and menopausal status. In conclusion, when reassuring women with insufficient sample on outpatient endometrial biopsy, one can be confident about absence of pathology provided the hysteroscopic and sonographic endometrial assessment is consistent with endometrial atrophy.

Objective: To evaluate the risk of premalignant and malignant pathology among endometrial polyps., Design: Prospective cohort study., Setting: Minimal Access Surgical Training (MAST) center in a large teaching hospital., Methods: Among 248 patients seen in outpatient hysteroscopy clinic (1996-97), 62 had endometrial polyps. All patients had endometrial sampling for histological assessment. To determine the magnitude of malignant potential among polyps, we compared the pathological findings in polyps (cases) with non-polypoidal specimens (controls)., Results: Out of 62 polyps, histologically 53 (85.5%) were benign, seven (11.3%) had hyperplasia, and two (3.2%) were associated with malignancy. Hyperplasia was more frequent in endometrial specimens with polyps than in those without (11.3% vs 4.3%, p=0.04), but the incidence of carcinoma in the two groups was the same (3.2% vs 3.2%, p= 1.0)., Conclusion: In abnormal uterine bleeding, hyperplasia was, but cancer was not, more common in women with endometrial polyps compared to those without polyps.

Objective: To determine the accuracy of ultrasound scan in the diagnosis of endometrial hyperplasia and cancer in postmenopausal bleeding., Design: A prospective diagnostic accuracy study (1996-97)., Setting: Minimal access surgical training centers in two large teaching hospitals., Methods: Ultrasound scan and outpatient endometrial sampling were performed on 96 patients with postmenopausal bleeding. Patients unable to have these outpatient procedures had a formal inpatient hysteroscopy and curettage. Test performance characteristics were computed for ultrasound scan comparing its estimate of endometrial thickness with histologic diagnosis that served as a 'gold' standard., Outcome Measures: Accuracy of the ultrasonic endometrial thickness was estimated using sensitivity, specificity and predictive values for binary data. For multilevel data, the diagnostic accuracy was computed using likelihood ratios (LRs). An LR < decreased the probability that endometrial hyperplasia/cancer was present, whereas an LR > 1 increased the probability that such lesion was present., Results: Using endometrial thickness > or =4 mm, the sensitivity of ultrasound to detect the endometrial malignancy was 92.9%, the specificity was 500%, and the positive and negative predictive values were 24.1% and 97.6% respectively. Analysis using likelihood ratio (LR) revealed that LR was 0.14 for endometrial thickness > or =4.0 mm, 0.94 for endometrial thickness 4.1-9.0 mm, and 3.3 for endometrial thickness >9.0 mm., Conclusion: In women with postmenopausal bleeding, malignancy can probably be safely excluded if sonographic endometrial thickness is < or = 4.0 mm. However, the probability of endometrial hyperplasia/cancer is not particularly altered by the knowledge that endometrial thickness on scan is >4.0 mm.