Imipenem-resistant Isolates Research Articles

Extended spectrum and metalo beta-lactamase producing airborne Pseudomonas aeruginosa and Acinetobacter baumanii in restricted settings of a referral hospital: a neglected condition

BackgroundFrequently encountered multidrug-resistant bacterial isolates of P. aeruginosa and A. baumannii are common and prevalent in a hospital environment. The aim of this study was to determine the prevalence and pattern of antibiotic resistance, extended spectrum and metallo beta-lactamase producing P. aeruginosa and A. baumannii isolates from restricted settings of indoor air hospital environment.MethodsA hospital-based cross-sectional study was conducted in Wolaita Sodo University Teaching and referral Hospital, Ethiopia from December 1/2015 to April 30/2015. The Air samples were collected from delivery room, intensive care unit and operation theatre of the hospital by active, Anderson six slate sampler technique during the first week of the months, twice a week during Monday’s and Friday’s. Standard microbiological procedures were followed to isolate P. aeruginosa and A. baumannii. Susceptibility testing was performed on isolates using the Kirby-Bauer disk diffusion technique. Extended spectrum beta lactamase production was detected by double disc synergy test and Imipenem-resistant isolates were screened for producing Metallo-beta lactamase.ResultsA total number of 216 indoor air samples were collected from the delivery room, intensive care unit, and operation room. Correspondingly, 43 A. baumannii isolates were identified (13 from delivery room, 21 from intensive care unit and 9 from operation room). Likewise 24 P. aeruginosa isolates were obtained (4 from delivery room, 13 from intensive care unit and 7 from operation room). Extended spectrum beta lactamase and metalo-beta lactamase production were observed in 24 (55.8%) and 13 (30.2%) isolates of A. baumannii respectively, whereas P. aeruginosa showed 15 (62.5%) extended spectrum beta lactamase and 9 (37.5%) metallo-beta lactamase production.ConclusionsExtended spectrum beta lactamase and metallo-beta lactamase producing bacteria in hospital air is a new dimension for specific setting of the study area where antimicrobial resistance is increasing and surgical site infection is prevalent. So, identification of these microorganisms has a great role in reducing the burden of antibiotic resistance and could also provide a significant input for framing hospital infection control policies.

Emerge of blaNDM-1 and blaOXA-48-like harboring carbapenem-resistant Klebsiella pneumoniae isolates from hospitalized patients in southwestern Iran

BackgroundOne of the most important emerging carbapenem-resistant bacteria is Klebsiella pneumoniae (K. pneumoniae). The present study aimed to investigate the antibiotic susceptibility pattern of K. pneumoniae isolates and detection of carbapenemase producing K. pneumoniae obtained from Iranian hospitalized patients. MethodsThis cross-sectional study was performed on 211 K. pneumoniae isolates which were recovered from different clinical specimens from 2014 to 2015. Modified Hodge test (MHT) and double disk synergy test (DDST) were done for detection of carbapenemase and metallo-beta-lactamase (MBL) producing K. pneumoniae. The presence of antibiotic resistance determinants was investigated by polymerase chain reaction (PCR) method. ResultsThe results of antibiotic susceptibility showed that all isolates were resistant to ampicillin, and then mostly resistant to piperacillin and ceftazidime with 76.3% and 66.8%, respectively. On the other hand, the highest sensitivity was toward polymyxin B, followed by carbapenems. Of 29 carbapenem-resistant isolates, all were high-level imipenem-resistant isolates (Minimum inhibitory concentration ≥4), except 4 isolates. The results of MHT and DDST showed that 93.1% (27/29) of carbapenem-resistant isolates were carbapenemase and MBL producing isolates, respectively. The presence of blaNDM-1 and blaOXA-48-like genes was detected in 27 (10.9%) and 2 (0.9%) isolates, respectively. ConclusionThis is the first identification of blaNDM-1 and blaOXA-48-like in K. pneumoniae in Southwestern Iran and the highest reported prevalence of blaNDM in this bacterium from Iran. Since carbapenem-resistant isolates containing New Delhi metallo-beta-lactamase 1 (NDM-1) were almost resistant to all the tested antibiotics, the resistance due to this gene may be increased in the near future as a potential health threat.

Evaluation of different phenotypic tests for detection of metallo-β-lactamases in imipenem-resistant Pseudomonas aeruginosa.

PURPOSE:Pseudomonas aeruginosa causes a wide spectrum of infections including bacteremia, pneumonia, urinary tract infection, etc., Metallo-beta-lactamase (MBL) producing P. aeruginosa is an emerging threat and cause of concern as they have emerged as one of the most feared resistance mechanisms. This study was designed to know the prevalence of MBL production in P. aeruginosa and to evaluate the four phenotypic tests for detection of MBL production in imipenem-resistant clinical isolates of P. aeruginosa.METHODS:Totally, 800 isolates of P. aeruginosa isolated from various clinical samples were evaluated for carbapenem resistance and MBL production. All imipenem-resistant strains were tested for carabapenemase production by modified Hodge test. Screening for MBL production was done by double-disc synergy test and combined disc test (CDT). Confirmation of MBL production was done by the E-test (Ab BioDisk, Solna, Sweden).RESULTS:Out of the 800 isolates of P. aeruginosa, 250 isolates were found resistant to imipenem. Based on the results of E-test, 147 (18.37%) isolates of P. aeruginosa were positive for MBL production. The CDT has the highest sensitivity and specificity for the detection of MBL production as compared to other tests.CONCLUSION:The results of this study are indicative that MBL production is an important mechanism of carbapenem resistance among P. aeruginosa. Use of simple screening test like CDT will be crucial step toward large-scale monitoring of these emerging resistant determinants. Phenotypic test for MBL production has to be standardized, and all the isolates should be routinely screened for MBL production.

Microbiological characteristics of hypermucoviscous Klebsiella pneumoniae isolates from different body fluids.

Reports of hypermucoviscous Klebsiella pneumoniae (hvKP) isolated from fluids other than blood or abscess are rare. The aim of the study was to compare clinical and microbiological characteristics of hvKP found in blood or abscess fluid with those isolated from other loci. A total of 24 non-repetitive hvKP isolates were collected from January 2013 to June 2014 from patients with hvKP infections. There were 15 in Group 1 (fluid other than blood or abscess) and 9 in Group 2 (blood or abscess fluid). Medical records of all patients were reviewed. Capsular polysaccharide (CPS) typing, virulence factor determination, and multilocus sequence typing (MLST) of hvKP isolates were performed. Seventeen sequence types (STs) and 6 capsular serotypes were identified. Type K2CC65 was most commonly identified in Group 1 and type K2CC86 in Group 2. Deletion of pLVPK-derived loci were found in K2 and non-K1/K2 hvKP strains. Two virulent genes, fimH and ycfM, were identified more frequently in Group 2 than in Group 1. There was no difference in the frequency of other virulent genes or serotypes in the two groups. Two imipenem resistant hvKP isolates (cr-hvKP) were found in non-blood or abscess samples. hvKP isolated from different body fluids had similar clinical and microbiological characteristics. cr-hvKP identified in non-blood or abscess samples should raise our attention to the challenging situation and management of hvKP infection.

Imipenem-resistant Gram-negative bacterial isolates carried by persons upon medical examination in Korea.

Carbapenem-resistant Gram-negative bacteria (CR-GNB) have emerged and disseminated worldwide, become a great concern worldwide including Korea. The prevalence of fecal carriage of imipenem-resistant Gram-negative bacteria (IR-GNB) in persons in Korea was investigated. Stool samples were collected from 300 persons upon medical examination. Samples were screened for IR-GNB by using MacConkey agar with 2 μl/ml imipenem. Species were identified by 16S rRNA gene sequence analysis, and antimicrobial susceptibility was determined by the broth microdilution method. In total, 82 IR-GNB bacterial isolates were obtained from 79 (26.3%) out of 300 healthy persons. Multilocus sequence typing analysis showed very high diversity among IR P. aeruginosa, S. maltophilia, and E. cloacae isolates, and pulsed-field gel electrophoresis revealed five main pulsotypes of IR P. mirabilis. As for the presence of metallo-β-lactamases (MBLs), only one IMP-25-producing S. marcescens isolate was identified. Although only one carbapenemase-producing isolate was identified, the high colonization rates with IR-GNB isolates in this study is notable because carriers may be a reservoir for the dissemination of resistant pathogens within the community as well as in health care institutions.

Multidrug resistant Stenotrophomonas maltophilia: An emerging cause of hospital acquired infections in Assiut University Hospitals, Egypt

Stenotrophomonas maltophilia is an emergent pathogen in health¬care facilities worldwide that is intrinsically resistant to many antibiotics. We aimed to determine the prevalence of S. maltophila causing health care associated infections (HAIs) and environmental contamination at the intensive care units (ICUs) of Assiut University Hospitals, the antibiotic resistance profiles, production of metallo-β-lactamases (MBLs) and detection of sul 2 gene. A total of 690 clinical samples and 4151 environmental samples were collected. S. maltophila isolates were identified and confirmed by detection of 16S rRNA-23S rRNA gene by PCR. Antimicrobial resistance was determined by Kirbey Bauer disc diffusion method and imipenem MIC by E test. The presence of MBLs were detected by combined disc test (CDT) and double disc synergy test (DDST). The presence of sul 2 gene was determined by PCR. S. maltophila caused 9.7% of HAIs in the ICUs mostly respiratory tract infections and 0.67% of environmental contamination . A high percentage of resistance was found for most of the studied antimicrobials including carbapenems. MBLs were detected in all imipenem resistant isolates and in variable percentages in imipenem sensitive isolates. The least resistance found was to trimethoprim sulfamethoxazole (SXT). All SXT resistant isolates harboured the sul 2 gene. Multidrug resistance was reported in 63.5% of isolates. S. maltophilia causes a considerable percentage of HAIs especially respiratory tract infections and environmental contamination in the ICUs. A high percentage is MDR. Trimethoprim-sulfamethoxazole is the single agent of choice for the treatment of S. maltophilia infections. The detection of sul2 gene and MBLs among isolates at our hospitals is alarming.

Antimicrobial Effects of β-Lactams on Imipenem-Resistant Ceftazidime-Susceptible Pseudomonas aeruginosa.

We studied the resistance mechanism and antimicrobial effects of β-lactams on imipenem-resistant Pseudomonas aeruginosa isolates that were susceptible to ceftazidime as detected by time-kill curve methods. Among 215 P. aeruginosa isolates from hospitalized patients in eight hospitals in the Republic of Korea, 18 isolates (23.4% of 77 imipenem-resistant isolates) were imipenem resistant and ceftazidime susceptible. Multilocus sequence typing revealed diverse genotypes, which indicated independent emergence. These 18 isolates were negative for carbapenemase genes. All 18 imipenem-resistant ceftazidime-susceptible isolates showed decreased mRNA expression of oprD, and overexpression of mexB was observed in 13 isolates. In contrast, overexpression of ampC, mexD, mexF, or mexY was rarely found. Time-kill curve methods were applied to three selected imipenem-resistant ceftazidime-susceptible isolates at a standard inoculum (5 × 105 CFU/ml) or at a high inoculum (5 × 107 CFU/ml) to evaluate the antimicrobial effects of β-lactams. Inoculum effects were detected for all three β-lactam antibiotics, ceftazidime, cefepime, and piperacillin-tazobactam, against all three isolates. The antibiotics had significant killing effects in the standard inoculum, but no effects in the high inoculum were observed. Our results suggest that β-lactam antibiotics should be used with caution in patients with imipenem-resistant ceftazidime-susceptible P. aeruginosa infection, especially in high-inoculum infections such as endocarditis and osteomyelitis.

Emergence of colistin resistance in Pseudomonas aeruginosa ST235 clone in South Korea

In this study, the prevalence and characteristics of metallo-β-lactamase (MBL)-producing Pseudomonas aeruginosa isolates in South Korea were investigated. Among 215 P. aeruginosa isolates collected from eight hospitals, 77 (35.8%) and 72 (33.5%) were resistant to imipenem and meropenem, respectively. Of the 77 imipenem-resistant isolates, MBL genes were identified in 34 isolates (blaIMP-6 in 33 isolates and blaVIM-2 in 1 isolate). All of the MBL-producing isolates belonged to a globally prevailing genotype, sequence type 235 (ST235), and all of the IMP-6-producing isolates showed a deletion of nucleotide 209 of the porin gene oprD. Of the 33 IMP-6-producing ST235 isolates, 9 were resistant to colistin and exhibited resistance to all antimicrobial agents included in this study. PhoPQ and PmrAB amino acid alterations were not identical in the colistin-resistant isolates, indicating independent emergence of colistin resistance in this high-risk clone. Carbapenem resistance in P. aeruginosa has increased in South Korea owing to the dissemination of IMP-6-producing ST235 isolates, which showed high-level resistance to meropenem. Emergence of colistin resistance in the disseminated resistant clone would be a significant threat because few alternatives are left for the treatment of systemic infections.

Molecular detection of blaNDM-1 (New Delhi metallobetalactamase-1) in nosocomial Enterobacteriaceae isolates by nested, multiplex polymerase chain reaction

BackgroundCarbapenems are considered “drugs of last resort” in many life-threatening infections. Advent of carbapenemases like KPC, OXA-48, VIM, IMP, and NDM have greatly affected the efficacy of these drugs, posing serious threat to global health and infection control. NDM bears special significance to the India subcontinent, labeled as place of origin and reservoir. NDM tends to escape detection by routine phenotypic methods, requiring molecular confirmation. This study utilizes nested, multiplex polymerase chain reaction (PCR) for reliable detection of blaNDM-1 in nosocomial Enterobacteriaceae isolates. MethodsThis study was conducted to detect prevalence of blaNDM-1, blaIMP, blaVIMand blaKPC genes by multiplex PCR among multidrug/carbapenem-resistant nosocomial Enterobacteriaceae isolates. From March 2013 to April 2014, 100 consecutive non-repeat isolates of Enterobacteriaceae from various inpatient clinical samples were analyzed. Imipenem-resistant isolates identified by Kirby Bauer disk diffusion method with Clinical and Laboratory Standards Institute guidelines were further subjected to nested, multiplex PCR to simultaneously detect blaNDM-1, blaIMP, blaVIMand blaKPC genes. ResultsOut of 100 isolates, 17 (17%) were found to be imipenem-resistant. blaNDM-1 was detected in all 17 isolates by nested, multiplex PCR. blaVIM was co-carried in 4 isolates while one isolate co-harbored blaIMP with blaNDM-1. Imipenem resistance and NDM-1 carriage was predominant amongst Klebsiella isolates. Maximum NDM-1 producers were isolated from the intensive care unit (70.6%). ConclusionNDM-1 prevalence in nosocomial Enterobacteriaceae isolates in our hospital was found to be 17%. A nested, multiplex PCR was used for rapid detection of various carbapenemase genes with high sensitivity and specificity which is essential not only for favorable patient outcome but also for timely implementation of appropriate infection control practices to prevent further spread of such organisms.

In vitro efficacy of synergistic antibiotic combinations in imipenem resistant Pseudomonas aeruginosa strains

The increase in antibiotic resistance coincided with the decline in production of new antibiotics. Combination antibiotic treatment is preferred in nosocomial infections caused by multidrug resistant Pseudomonas aeruginosa. In vitro synergism test by agar dilution method were used to choose the combinations which might be used in clinic. The aim of this study was to investigate the synergistic efficacy of antibiotic combinations in imipenem resistant P. aeruginosa strains. Carbapenem resistance (imipenem and meropenem) wasdetermined by disk diffusion method. Among isolated P. aeruginosa 44.9% were cabapenem resistant. The MIC of drugs among 25 imipenem resistant isolates ranged from >_ 256 mg/L to <_ 8 mg/L for imipenem, >_ 1024 mg/L to <_ 64 mg/L for ceftriaxone, >_ 256 mg/L to <_ 8 mg/L for amikacin, >_ 16 mg/L to <_ 2 mg/L for colistin, >_ 512 mg/L to <_ 16 mg/L for piperacillin/tazobactam. Among antibiotic combinations, piperacillin /tazobactam- amikacin was most effective with 80% synergism next to which was imipenem-amikacin with 60% synergism, then imipenem-colistin with 50% synergism, imipenem-ceftriaxone with 30% synergism. Only one combination (piperacillin/tazobactum -imipenem showed 20% antagonism. All these combinations had considerable proportion of additive effect which is also desirable for these multi drug resistant isolates.Bangladesh J Med Microbiol 2015; 9 (1): 3-8

Phenotypic and molecular characterization of clinical Acinetobacter isolates from Menoufia University Hospitals

Objectives The aims of this study were to investigate the prevalence of Acinetobacter spp. in Menoufia University Hospitals, to investigate their antimicrobial susceptibility patterns, and to assess carbapenemases production in these isolates. Background Acinetobacter spp. are important opportunistic pathogens responsible for nosocomial infections. Materials and methods This study was conducted on 603 clinical samples from patients admitted to Menoufia University Hospitals. Acinetobacter spp. were identified by standard microbiological methods and API20NE test kits. Antimicrobial susceptibility was tested using disk diffusion and agar dilution methods. Imipenem-resistant Acinetobacter isolates were further tested for metallo-β-lactamase (MβL) production. Results This study was conducted at Medical Microbiology and Immunology Laboratory, Faculty of Medicine, Menoufia University. Acinetobacter spp. represented 10.6% of all collected nosocomial isolates. With regards to API20NE results, A. baumanii was the predominant Acinetobacter spp. (80.8%) followed by A. baumannii–A. calcoaceticus complex (7.7%), A. lwoffii (5.8%), A. haemolyticus (3.8%), and A. pitti (2.6%). Acinetobacter isolates were highly resistant to cefepime (92.3%), ampicillin–sulbactam, piperacillin, piperacillin–tazobactam, ceftazidime, tobramycin (91% for each), amikacin (84.6%), and imipenem (67.9%). Overall, 56.4% of Acinetobacter isolates were susceptible to tigecycline. On agar dilution method, 96.2% of Acinetobacter isolates were found to be susceptible to colistin and 66.7% were imipenem resistant. Imipenem/ethylenediaminetetraacetic acid combined disk test showed that 81.1% of imipenem-resistant Acinetobacter were MβL producers, and multiplex PCR showed that 15.1% of imipenem-resistant Acinetobacter were positive for blaVIM2, but none of them were positive for blaIMP1 gene. Conclusion Acinetobacter spp. are serious nosocomial pathogens with high prevalence of carbapenems resistance. Production of carbapenemases, especially MβLs, is considered the main carbapenem-resistance mechanism. Tigecycline and colistin can be valuable therapeutic options for the treatment of Acinetobacter infections.

Investigation of In Vitro Activity of Mecillinam Against Isolates of Enterobacteriaceae

Introduction: Mecillinam is an oral antibiotic with excellent clinical efficacy in the treatment of uncomplicated UTIs. It is a betalactam antibiotic and active against Enterobactericeae but has little activity against Gram positive bacteria. In this study, we aimed to investigate the in vitro activity of mecillinam against Enterobactericeae isolates that were isolated from different clinical specimens. Materials and methods: Identification of the bacterial isolates were tested by conventional methods and Phoenix (Becton Dickinson, USA) and Vitek MS (Biomeriux, France) automated systems. Antimicrobial susceptibility was performed by using the disk diffusion method and Phoenix (Becton Dickinson, USA) and Vitek2 Compakt (Biomeriux, France) autometed systems. Mecillinam susceptibility of the isolates were determined by disc diffusion method by using 10 μg mecillianm disc and interpreted according to the EUCAST criteria. Results: Total of 338 Enterobactericeae (E. coli n=162, Klebsiella spp. n=142, Enterobacter spp. n=18, Proteus spp. n=10, Citrobacter spp. n=4, Salmonella spp. n=2) isolates were tested. Most of the clinical specimens were urinary specimens (n=272, 80.5%). Mecillinam susceptibility was detected as 82.5%. Mecillinam susceptibility for urinary tract isolates was 87.1%. In this study, 57% of imipenem resistant isolates were determined susceptible for mecillinam. Conclusion: In conclusion, Mecillinam could be a good option, especially for urinary tract infections.

Molecular characterization and resistance profile of nosocomial Acinetobacter baumannii intensive care unit of tertiary care hospital in Bangladesh.

This study was designed to investigate the resistance profile along with the genetic background of resistance to beta-lactam antibiotics among the nosocomial A. baumannii in Bangladesh. A. baumannii was confirmed by detecting blaoXA-51-like. Antibiotic susceptibility was determined by disk diffusion method. Agar dilution method was used to determine MIC of ceftazidime and imipenem. All A. baumannii were phenotypically screened for ampC, ESBL and MBL production. Genetic markers of antibiotic resistance. such as blaampC, blaOXA-51, 23, 40, 58 and 143, blaKPc, blaMp, blavi and blaNDM-j, genetic environment around blaADc and ISAbal upstream of blaoXA, were evaluated by PCR. Twenty-four (96%) A. baumannii were considered as MDR. 96% A. baumanii were resistant to amoxiclav, ceftazidime, ciprofloxacin and cefoxitin, 92% to cefotaxime and piperacillin-tazobactam, 88% to cefepime, amikacin and imipenem, 52% to sulbactam- cefoperazone and 40% were resistant to aztreonam. Everything were sensitive to colistin. The distribution of several beta-lactamase genes such as blaoxa-51 (100%), blaADC-like (92%), blaNDM-i (92%), EBC group (84%), blaoxa-23 (76%), blavm (72%), blacpc (44%), DHA group (24%), blaoxa-58 (16%), ACC group (8%) and CIT group (4%) were observed among the 25 A. baumannii. This is the first reported plasmid mediated ampC beta-lactamases in A. baumannii. blaoxa-51 was positive in 100%, blandm-i in 95.45%, blaoxa-23 in 77.27%, blavim in 72.73%, blakpc in 50% and blaOXA-58 in 18.18% of imipenem resistant isolates. MDR profile of nosocomial A. baumannii would highlight the importance of standard guideline of antimicrobials use and infection control policy in the hospitals of Bangladesh.

Detection of ISPa1328 and ISPpu21, Two Novel Insertion Sequences in the OprD Porin and blaIMP-1 Gene Among Metallo-Beta-Lactamase-Producing Pseudomonas aeruginosa Isolated From Burn Patients

Background Carbapenemes are a good choice for treatment of infections caused by multidrug resistant Pseudomonads aeruginosa. The emergence of carbapenem resistance has become a major problem in treatment of this organism especially among immunocompromised patients including burn patients. Objectives The aim of this study was to investigate carbapenem-resistance mechanisms among burn patients in Tehran, Iran, during 2014 - 2015. Methods The antibiotic resistance phenotypic test was accomplished by the Kirby Bauer disk diffusion method. The phenotypic investigation of metallo-beta-lactamase (MBL) producers was evaluated by the combined disk diffusion test (CDDT) method. The prevalence of MBL genes, including blaIMP-1 and blaVIM-1 was evaluated by polymerase chain reaction (PCR) and sequencing methods. Amplification of oprD was performed by PCR and the results of sequencing were aligned with wild-type P. aeruginosa strain PAO1. Results A total of 100 P. aeruginosa were investigated, of which, 95 were resistance to imipenem. Out Of 95 imipenem resistant isolates,, 81 (85.2%) were MBL producers. Among all isolates, 13 strains carried the blaIMP-1 gene, whereas all of the strains were negative for the blaVIM-1 gene. Amplification of OprD porin was performed for all 100 P. aeruginosa strains. Two insertion sequences (ISs) including ISPpu21 and ISPa1328 were detected in PCR products of OprD gene, that were larger than expected. Conclusions The prevalence of β-lactamase-producing isolates and their isolation from life-threatening infections in burn patients is increasing at an alarming rate worldwide. Also, we have identified two novel IS elements, ISPa1328 and ISPpu21, in P. aeruginosa isolates from hospitals in Tehran, Iran. In most of the isolates, insertional inactivation of oprD by ISPa1328 and ISPpu21 were associated with carbapenem resistance.

Resistant mechanisms and molecular epidemiology of imipenem-resistant Acinetobacter baumannii.

The aim of the study was to investigate the resistant mechanisms and homology of imipenem-resistant Acinetobacter baumannii (A. baumannii). A total of 46 non-duplicate imipenem-resistant A. baumannii clinical isolates were collected from three tertiary hospitals between July, 2011 and June, 2012. The minimal inhibitory concentrations (MICs) of antimicrobial agents were determined using the agar dilution method. Phenylalanine-arginine β-naphthylamide was used to detect the presence of the efflux pump-mediated resistant mechanism. Polymerase chain reaction was employed to amplify genes associated with drug resistance, including β-lactamase genes, efflux pump genes and outer membrane protein gene CarO. A few amplicons were randomly selected and sequenced. Multilocus sequence analysis (MLST) was employed in typing A. baumanni. A. baumannii was resistant to imipenem, simultaneously showing resistance to several other antimicrobials. In addition, 13 A. baumannii were found to mediate drug resistance through operation of the efflux pump. Of the various drug resistance genes tested, blaOXA-51 was present in 46 isolates, blaOXA-23 gene was present in 44 isolates and blaNDM gene was found in only one strain. Other drug resistant-associated genes, including blaKPC, blaIMP, blaOXA-24, blaOXA-58, blaSHV, blaGIM and blaVIM were not detected. Mutation of adeS and outer membrane protein gene CarO were found in a few of the imipenem-resistant isolates. The MLST analysis revealed that all 46 clinical isolates were clustered into 11 genotypes and the most frequent genotype was ST208. In conclusion, β-lactamase genes, genes involved in efflux pump and mutation of outer membrane protein encoding gene may be important in mediating imipenem resistance in A. baumannii. Of the 11 different genotypes, ST11 was shared by the majority of A. baumannii, which may be due to horizontal transfer of patients from hospitals.

Antibiotic resistance patterns of Gram-negative and Gram-positive strains isolated from inpatients with nosocomial infections in a tertiary hospital in Beijing, China from 2011 to 2014

This study was to evaluate the resistance of antimicrobial agents against pathogens from inpatients with nosocomial infection collected in Beijing, China, during 2011–2014. Measurement of minimum inhibitory concentrations (MICs) was carried out using the broth microdilution method with the Clinical and Laboratory Standards Institute (CLSI) guidelines as the indicator. A total of 5442 Gram-negative and 806 Gram-positive isolates were collected in this study in 2011–2014. Two carbapenem-resistant strains appeared among Escherichia coli (E. coli), while imipenem-resistant isolates increased in proportion from 0% to 8.2% among Klebsiella pneumonia (K. pneumonia) during 4 year. Acinetobacter baumannii (A. baumannii) revealed severe antibacterial resistance to most antimicrobial agents. In contrast, a decreasing trend on resistance had been observed among Pseudomonas aeruginosa (P. aeruginosa) especially after 2012, range from 1.8% for co-trimoxazole to 13.5% for piperacillin. The resistance of Staphylococcus aureus (S. aureus) also had the lowest resistant to linezolid and vancomycin (0.1%). In summary, antimicrobial-resistant nosocomial pathogens have gradually increased from 2011 to 2014, so improved surveillance of hospital-acquired infections and effective infection-control measures may be the best way to solve the present problem.

High prevalence of blaVIM-1 gene in bacteria from Brazilian soil.

This study investigated bacteria from soil samples to (i) determine the main bacterial genera and species having resistance to carbapenem and other β-lactams and (ii) establish if the mechanism of resistance was due to the production of metallo-β-lactamases. The isolates were characterized by PCR for metallo-β-lactamases and integrons, by antimicrobial susceptibility testing, and by sequencing. The antimicrobial profile of 40 imipenem-resistant Gram-positive soil isolates from all Brazilian regions demonstrated that 31 (77.5%) of them were multidrug resistant. Among the 40 isolates, 19 presented the blaVIM gene and class 1 integrons by PCR. Six of the 19 isolates were identified as Paenibacillus sp., 12 as Bacillus sp., and just 1 was classified as Staphylococcus sp., by sequencing of the 16S rRNA gene. These results suggest that bacteria from soil can act as a source of blaVIM-1 genes, representing a threat to public health.

Early detection of metallo-β-lactamase NDM-1- and OXA-23 carbapenemase-producing Acinetobacter baumannii in Libyan hospitals

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.

Antibiotic Resistance Patterns and a Survey of Metallo-β-Lactamase Genes Including bla-IMP and bla-VIM Types in Acinetobacter baumannii Isolated from Hospital Patients in Tehran

Background: Metallo-β-lactamases (MBLs) producing strains of Acinetobacter baumannii are serious etiological agents of hospital infections worldwide. Among the β- lactams, carbapenems are the most effective antibiotics used against A. baumannii. However, resistance to these drugs among clinical strains of A. baumannii has been increasing in recent years. In this study, the antimicrobial sensitivity patterns of A. baumannii strains isolated from eleven different hospitals in Tehran, Iran, and the prevalence of MBL genes (bla-_VIM and bla-_IMP) were determined. Method: During a period of 5 months, 176 isolates of A. baumannii were collected from different clinical specimens from hospitalized patients in Tehran. All isolates were confirmed by biochemical methods. The isolates were tested for antibiotic sensitivity by the Kirby-Bauer disk diffusion method. Following minimum inhibitory concentration determination, imipenem-resistant isolates were further tested for MBL production by the double disk synergy test (DDST) method. PCR assays were performed for the detection of the MBL genes bla-_IMP and bla-_VIM. Results: The DDST phenotypic method indicated that among the 169 imipenem-resistant isolates, 165 strains were MBL positive. The PCR assays revealed that 63 of the overall isolates (36%) carried the bla-_VIM gene and 70 strains (40%) harbored bla-_IMP. Conclusions: It is obvious that nosocomial infections associated with multidrug-resistant Acinetobacter spp. are on the rise. Therefore, the determination of antibiotic sensitivity patterns and screening for MBL production among A. baumannii isolates is important for controlling clinical Acinetobacter infections.

Insertion sequence ISRP10 inactivation of the oprD gene in imipenem-resistant Pseudomonas aeruginosa clinical isolates

Carbapenem resistance mechanisms were investigated in 32 imipenem-resistant Pseudomonas aeruginosa clinical isolates recovered from hospitalised children. Sequence analysis revealed that 31 of the isolates had an insertion sequence element ISRP10 disrupting the porin gene oprD, demonstrating that ISRP10 inactivation of oprD conferred imipenem resistance in the majority of the isolates. Multilocus sequence typing (MLST) was used to discriminate the isolates. In total, 11 sequence types (STs) were identified including 3 novel STs, and 68.3% (28/41) of the tested strains were characterised as clone ST253. In combination with random amplified polymorphic DNA (RAPD) analysis, the imipenem-resistant isolates displayed a relatively high degree of genetic variability and were unlikely associated with nosocomial infections.