scholarly journals oprD Genes Detected in Pseudomonas aeruginosa Isolates from a Teaching Hospital but Lost in a Carbapenem-Resistant Strain

2019 ◽  
pp. 1-8 ◽  
Author(s):  
Eucharia E. Nmema ◽  
Chioma S. Osuagwu ◽  
Eunice N. Anaele
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Teaching Hospital ◽  
Multiple Drug Resistance ◽  
Carbapenem Resistance ◽  
University Teaching ◽  
Diffusion Method ◽  
Multiple Drug ◽  
Disk Diffusion Method ◽  
Chain Reactions ◽  
Carbapenem Resistant

Aims: The aims of the study were to evaluate the multidrug resistance profile and mechanisms of carbapenem resistance in Pseudomonas aeruginosa clinical isolates using phenotypic and genotypic methods. Study Design: A descriptive laboratory based study. Place and Duration of Study: Microbiology Laboratory, Ondo State University of Science and Technology, Okitipupa, and Biotechnology Laboratory, Ladoke Akintola University of Technology, Osogbo, Nigeria, between June 2017 and November 2018. Methodology: Ten P. aeruginosa isolates were recovered from patients at Lagos University Teaching Hospital, and susceptibilities to imipenem (10 µg), meropenem (10 µg) and a panel of antibiotics were performed by the disk diffusion method. Genotypic methods including Polymerase Chain Reactions (PCR) and agarose gel electrophoresis were carried out according to established protocols. oprD and blaIMP gene primers were used for the PCR amplification. Results: Fifty percent (50%) of the isolates showed multiple drug resistance. Four isolates (40%) were carbapenem resistant (CR). oprD gene was detectedin 90% (9/10) of the isolates. 75% (3/4) of CR strains were among the strains showing oprD gene. 25% (1/4) CR strain (PA1421) was oprD negative. Loss or mutation of oprD gene seems to be the mechanism of carbapenem resistance in strain PA1421. Conclusion: Loss or mutation of oprD gene was identified in this study as a mechanism of carbapenem resistance. oprD gene encodes the outer membrane protein (OprD) porin in P. aeruginosa whose deficiency confers resistance to carbapenems, especially imipenem. Surveillance of the antimicrobial susceptibility patterns of P. aeruginosa is of critical importance in understanding new and emerging resistance trends, reviewing antibiotic policies and informing therapeutic options.

scholarly journals Prevalence of carbapenemase production in Pseudomonas aeruginosa isolates causing clinical infections in Lagos University Teaching Hospital, Nigeria

2021 ◽  
Vol 22 (4) ◽  
pp. 498-503
Author(s):  
A.O. Ettu ◽  
B.A. Oladapo ◽  
O.O. Oduyebo
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Teaching Hospital ◽  
Dipicolinic Acid ◽  
Phenylboronic Acid ◽  
Carbapenem Resistance ◽  
Clinical Isolates ◽  
University Teaching ◽  
Diffusion Method ◽  
Carbapenem Resistant ◽  
Mueller Hinton

Background: Pseudomonas aeruginosa has been highly associated with carbapenem resistance in which carbapenemases has been suggested to be a major contributory factor. Hence the objective of this study was to phenotypically detect KPC-type carbapenemase, metallo-β-lactamase and OXA-48 carbapenemase production in clinical isolates of P. aeruginosa in Lagos University Teaching Hospital (LUTH), NigeriaMethodology: One hundred and seventy-one P. aeruginosa isolates consecutively recovered from clinical specimens of patients with infections at the Medical Microbiology and Parasitology laboratory of the hospital were identified using MicrobactTM 24E kit. Preliminary screening for carbapenem resistance was determined by the disc diffusion method on Mueller-Hinton agar using single discs of meropenem and imipenem. Phenotypic detection of carbapenemase production among carbapenem-resistant isolates was performed by the combination disc test of meropenem-phenylboronic acid (MRPBO) and meropenem-dipicolinic acid (MRPDP) as recommended by EUCAST 2013 guideline. Results: Out of the 171 P. aeruginosa isolates, 35 (20.5%) were carbapenem non-susceptible (resistant) while carbapenemase production was detected in 27 (77.1%) of these carbapenem resistant isolates, and no enzyme was detected in 8 (22.9%). Of the 27 carbapenemase producing isolates, 22 (81.5%) produced MBL, 1 (3.7%) produced KPC, while 4 (14.8%) produced both KPC and MBL enzymes. Conclusion: This study revealed that carbapenem resistance among P. aeruginosa clinical isolates in our institution is gradually increasing. The mechanism for this rise is associated with carbapenemases, with MBL being the major carbapenemase involved. There is the need to ensure strict compliance with the LUTH infection control guidelines in order to check the rising incidence of infection caused by carbapenem resistant P. aeruginosa.   French title: Prévalence de la production de carbapénémases dans les isolats de Pseudomonas aeruginosa causant des infections cliniques à l'hôpital universitaire de Lagos, Nigéria   Contexte: Pseudomonas aeruginosa a été fortement associé à la résistance aux carbapénèmes dans laquelle les carbapénèmases ont été suggérées comme étant un facteur contributif majeur. Par conséquent, l'objectif de cette étude était de détecter phénotypiquement la production de carbapénémase de type KPC, de métallo-β-lactamase et de carbapénémase OXA-48 dans des isolats cliniques de P. aeruginosa au Lagos University Teaching Hospital (LUTH), Nigeria. Méthodologie: Cent soixante et onze isolats de P. aeruginosa récupérés consécutivement à partir d'échantillons cliniques de patients infectés au laboratoire de microbiologie médicale et de parasitologie de l'hôpital ont été identifiés à l'aide du kit MicrobactTM 24E. Le dépistage préliminaire de la résistance aux carbapénèmes a été déterminé par la méthode de diffusion sur disque sur gélose Mueller-Hinton en utilisant des disques uniques de méropénème et d'imipénème. La détection phénotypique de la production de carbapénèmes parmi les isolats résistants aux carbapénèmes a été réalisée par le test de disque combiné d'acide méropénème-phénylboronique (MRPBO) et d'acide méropénème-dipicolinique (MRPDP) tel que recommandé par la directive EUCAST 2013. Résultats: Sur les 171 isolats de P. aeruginosa, 35 (20,5%) étaient des carbapénèmes non sensibles (résistants) tandis que la production de carbapénèmes a été détectée dans 27 (77,1%) de ces isolats résistants aux carbapénèmes, et aucune enzyme n'a été détectée dans 8 (22,9%). Sur les 27 isolats producteurs de carbapénémases, 22 (81,5%) produisaient des MBL, 1 (3,7%) produisaient des KPC, tandis que 4 (14,8%) produisaient à la fois des enzymes KPC et MBL. Conclusion: Cette étude a révélé que la résistance aux carbapénèmes parmi les isolats cliniques de P. aeruginosa dans notre institution augmente progressivement. Le mécanisme de cette augmentation est associé aux carbapénémases, la MBL étant la principale carbapénémase impliquée. Il est nécessaire de garantir le strict respect des directives de contrôle des infections LUTH afin de contrôler l'incidence croissante des infections causées par P. aeruginosa résistant aux carbapénèmes.

Detection of Metalo-Beta-Lactamase Gene in Carbapenem Resistant Pseudomonas Aeruginosa Isolated From Lahore, Pakistan

2020 ◽  
Vol 3 (1) ◽  
Author(s):  
Humera Kausar ◽  
Shabbir Hussain ◽  
Afia Muhammad Akram
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Multiple Drug Resistance ◽  
Diffusion Method ◽  
Multiple Drug ◽  
Carbapenem Resistant ◽  
Routine Laboratory Examination ◽  
Polymerase Chain ◽  
Biochemical Testing ◽  
Resistance Rates ◽  
Lactamase Gene

Pseudomonas aeruginosa is a widespread organism, caused severe nosocomial infection in human andassociated with multiple drug resistance (MDR) Objective: The present study was carried out to observecurrent antimicrobial resistant pattern of Pseudomonas aeruginosa in Lahore and to detect the Metallobeta-lactamase (MBL) gene in carbapenem resistant Pseudomonas aeruginosa Methods: By screening360 samples total 123 Pseudomonas aeruginosa was identified by standard microbiology techniques suchas microscopy and biochemical testing. The isolated Pseudomonas aeruginosa was evaluated for drugresistance by disc diffusion method and polymerase chain reaction (PCR) was used to identify thecarbapenem resistance causing gene (bla-VIM and bla-IMP) Results: Following antibiotic resistantpattern was observed, Gentamycin (59.00%), Ceftazidime (58.7%), Ceftriaxone (58.00%), Cefotazime(57.0%) and Ciprofloxacin (55.00%). Resistance rates to carbapenem group of antibiotics is Doripenem(30.5%) Meropenem (31.0%) and Imipenem (28.0%). Out of 123 samples of Pseudomonas aeruginosa, 28isolates were found resistant to carbapenem group of antibiotic which was supposed to be highlysensitive for this bacterium. Molecular based identification of resistance genes showed that bla-IMP genewas present in 32.1% (09) and bla-VIM was found positive in 17.8% (04) samples. Metallo-beta-lactamasesproducing genes (bla-VIM and bla-IMP), among carbapenem resistant Pseudomonas aeruginosa weredetected in 28.1% of samples. If other carbapenem resistant gene were also included this number mightbe higher Conclusions: PCR based test should be included in routine laboratory examination for quickdetection of the resistance causing genes.

scholarly journals Detection of VIM and NDM-1 metallo-beta-lactamase genes in carbapenem-resistant Pseudomonas aeruginosa clinical strains in Bahrain

2019 ◽  
Vol 11 (02) ◽  
pp. 138-143 ◽  
Author(s):  
Ronni Mol Joji ◽  
Nouf Al-Rashed ◽  
Nermin Kamal Saeed ◽  
Khalid Mubarak Bindayna
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Multiple Drug Resistance ◽  
Infectious Agent ◽  
Carbapenem Resistance ◽  
Conventional Polymerase Chain Reaction ◽  
Control Measures ◽  
Multiple Drug ◽  
Beta Lactamase ◽  
Infection Control Measures ◽  
Carbapenem Resistant

Abstract INTRODUCTION: Carbapenem-resistant Pseudomonas aeruginosa has emerged as a life-threatening infectious agent worldwide. Carbapenemase genes are reported to be some of the most common mechanisms for carbapenem resistance in P. aeruginosa. No reports are available from the Kingdom of Bahrain about carbapenem resistance and the underlying cause. In this study, we determined to study the presence of the metallo-beta-lactamase (M β L) genes of VIM family and NDM-1 in carbapenem-resistant P. aeruginosa strains. METHODOLOGY: Fifty carbapenem-resistant P. aeruginosa isolates were obtained from three main hospitals of Bahrain. They were subjected to antimicrobial susceptibility testing by disc diffusion test. Subsequently, MβL was detected by imipenem-ethylene diamine tetraacetic acid (EDTA) combined disc test and conventional polymerase chain reaction. RESULTS: Among 50 P. aeruginosa strains, 40 (80%) were imipenem resistant. Among the 40 imipenem-resistant strains, 35 (87.5%) strains were positive for the imipenem-EDTA combined disc test, and 21 (52%) were carrying MβL genes. Nineteen (47.5%) strains were positive for the VIM gene; one (2.5%) strain was carrying the NDM-1 gene, while one strain was carrying both the VIM and NDM-1 genes. None of the imipenem sensitive strains carried the VIM or NDM-1 gene. CONCLUSION: This is the first study to report the presence of the VIM family gene and NDM-1 genes in imipenem-resistant P. aeruginosa isolates in the Kingdom of Bahrain. The study also confirms the multiple drug resistance by the MβL strains, attention should therefore from now on, be focused on prevention of further spread of such isolates by firm infection control measures, and to reduce its threat to public health.

scholarly journals New Delhi Metallo-β-lactamase (NDM)-mediated Carbapenem-Resistant Pseudomonas aeruginosa Clinical Isolates in Sudan

2018 ◽  
pp. 1-5
Author(s):  
Salma Elnour Rahma Mohamed ◽  
Alfadil Alobied ◽  
Mohamed Ibrahim Saeed ◽  
Wafa Mohamed Hussien
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Teaching Hospital ◽  
Carbapenem Resistance ◽  
Clinical Isolates ◽  
Diffusion Method ◽  
New Delhi ◽  
Army Hospital ◽  
Carbapenem Resistant ◽  
Wide Range ◽  
Data Documentation

Carbapenem resistance mediated by NDM is particularly gruesome as this carbapenemase can hydrolyze a wide range of β-lactam antibiotics. Aim: This study aims to detect NDM mediated carbapenem resistance in clinical isolates of Pseudomonas aeruginosa. Materials and Methods: 50 multi-drug resistant clinical urinary isolates of Pseudomonas aeruginosa from three major hospitals in Khartoum state Sudan; Khartoum Teaching Hospital, Medical Army Hospital and Omdurman teaching hospital, in period from July 2016 to September 2017, were investigated for carbapenem resistance using standard disc diffusion method and underwent real-time PCR to detect carbapenem resistance gene blaNDM. Data were analyzed using IBM SPSS. Results: 60% were positive for the blaNDM, 82% were resistant to Imipenem and 75% of the samples were resistant to Meropenem. Conclusion: The emergence of carbapenem resistance is a global problem that requires earnest attention. To make the suitable preventive measures, the emergence of these genes must be monitored closely. Our findings revealed that carbapenem-resistant due to the gene blaNDM is accounted for 60% of the cases, and due to lack of proper data documentation about the emergence of this gene in Sudan, these cases to the best of our knowledge are the first to be reported in Sudan.

scholarly journals First Report ofblaIMP-14on a Plasmid Harboring Multiple Drug Resistance Genes in Escherichia coli Sequence Type 131

2016 ◽  
Vol 60 (8) ◽  
pp. 5068-5071 ◽  
Author(s):  
Nicole Stoesser ◽  
Anna E. Sheppard ◽  
Gisele Peirano ◽  
Robert P. Sebra ◽  
Tarah Lynch ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Pseudomonas Aeruginosa ◽  
Multiple Drug Resistance ◽  
Carbapenem Resistance ◽  
Sequence Type ◽  
Multiple Drug ◽  
Class 1 Integron ◽  
Content Type ◽  
Carbapenem Resistant ◽  
Class 1

ABSTRACTTheblaIMP-14carbapenem resistance gene has largely previously been observed inPseudomonas aeruginosaandAcinetobacterspp. As part of global surveillance and sequencing of carbapenem-resistantEscherichia coli, we identified a sequence type 131 strain harboringblaIMP-14within a class 1 integron, itself nested within an ∼54-kb multidrug resistance region on an epidemic IncA/C2plasmid. The emergence ofblaIMP-14in this context in the ST131 lineage is of potential clinical concern.

scholarly journals The Bacteriological Quality, Safety, and Antibiogram ofSalmonellaIsolates from Fresh Meat in Retail Shops of Bahir Dar City, Ethiopia

2017 ◽  
Vol 2017 ◽  
pp. 1-5 ◽  
Author(s):  
Melkamnesh Azage ◽  
Mulugeta Kibret
Keyword(s):  
Drug Resistance ◽  
Multiple Drug Resistance ◽  
Disease Outbreaks ◽  
High Rate ◽  
Personal Hygiene ◽  
Diffusion Method ◽  
Multiple Drug ◽  
Bahir Dar ◽  
Disk Diffusion Method ◽  
Fresh Meat

The habit of raw meat consumption in addition to the poor hygienic standards and lack of knowledge contribute to food-borne diseases outbreaks. The objective of this research was to assess the bacterial quality and safety of fresh meat from retail Bahir Dar City, Ethiopia. A total of 30 fresh meat samples were collected from butcher shops. Standard bacteriological methods were used to isolate and enumerate bacteria. Kirby-Bauer disk diffusion method was used for antimicrobial susceptibility testing ofSalmonellaisolates. The mean counts of AMB, TC, andS. aureuswere log104.53, 3.97, and 3.88 log10cfu/g, respectively.Salmonellawas isolated from 21 (70%) of the samples.Salmonellaisolates in this study were highly susceptible to ciprofloxacin, gentamycin, and norfloxacin while they were resistant to erythromycin and tetracycline. High rate of multiple drug resistance was also noticed inSalmonellaisolates. The microbial loads of meat were above the recommended microbial safety limits. Besides this, the isolation rate ofSalmonellawas high and high levels of drug resistance were documented forSalmonellaisolates. Measures on handling and appropriate personal hygiene practices of workers in the retail shops are recommended to reduce the change of forborne disease outbreaks.

scholarly journals Phenotypic and genotypic characterization of multidrug-resistant isolates from patients with catheter-associated urinary tract infection in a tertiary care hospital

2019 ◽  
Vol 11 (03) ◽  
pp. 206-211
Author(s):  
Jaison Jayakaran ◽  
Nirupa Soundararajan ◽  
Priyadarshini Shanmugam
Keyword(s):  
Urinary Tract ◽  
Tertiary Care Hospital ◽  
Tertiary Care ◽  
Carbapenem Resistance ◽  
Multidrug Resistant ◽  
Diffusion Method ◽  
Care Hospital ◽  
Disk Diffusion Method ◽  
Carbapenem Resistant ◽  
Tract Infections

Abstract INTRODUCTION: Urinary tract infections (UTIs) remain as the most common infection. Catheter-associated (CA) UTI can lead to bacteremia and thereby is the leading cause of morbidity and mortality in hospitalized patients in our country. AIMS AND OBJECTIVES: This study aims to check the prevalence of CAUTI and study the phenotypic and genotypic characters of the multidrug-resistant organisms in a tertiary care hospital, with special reference to NDM-1 and OXA-23. MATERIALS AND METHODS: A total of 231 urine samples from patients with CA-UTI in different wards in a tertiary care hospital over a period of 3 months between June and August 2018 were collected and processed following the standard protocol. Antibiotic susceptibility tests were performed by disk-diffusion method. Modified Hodge test (MHT) was done to isolate carbapenem-resistant isolates, and polymerase chain reaction was done to detect NDM-1 and OXA-23. RESULTS: Out of 231 samples, 101 samples yielded significant growth. These 38 samples were Gram-negative bacilli which were resistant to carbapenems. Out of the 38 which showed carbapenem resistance, 23 were MHT positive. Out of the 23 MHT-positive isolates, 8 (21.05%) were positive for NDM-1 gene and only 1 (2.6%) was positive for the OXA-23 gene. CONCLUSION: This study has shown that carbapenem-resistant isolates from all the CA urinary tract-infected patients were 52.77% and most of them were Klebsiella. About 21% of them harbored the NDM-1 gene whereas only 2% had the OXA-23 gene. There has been an alarming increase in the spread of carbapenem resistance.

scholarly journals Occurrence of NDM-1 and VIM-2 Co-Producing Escherichia coli and OprD Alteration in Pseudomonas aeruginosa Isolated from Hospital Environment Samples in Northwestern Tunisia

Diagnostics ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 1617
Author(s):  
Raouaa Maaroufi ◽  
Olfa Dziri ◽  
Linda Hadjadj ◽  
Seydina M. Diene ◽  
Jean-Marc Rolain ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Pseudomonas Aeruginosa ◽  
Bacterial Resistance ◽  
Carbapenem Resistance ◽  
Multidrug Resistant ◽  
Hospital Environment ◽  
Diffusion Method ◽  
Resistant Bacteria ◽  
Multidrug Resistant Bacteria ◽  
Disk Diffusion Method

Hospital environments constitute the main reservoir of multidrug-resistant bacteria. In this study we aimed to investigate the presence of Gram-negative bacteria in one Northwestern Tunisian hospital environment, and characterize the genes involved in bacterial resistance. A total of 152 environmental isolates were collected from various surfaces and isolated using MacConkey medium supplemented with cefotaxime or imipenem, with 81 fermenter bacteria (27 Escherichia coli, and 54 Enterobacter spp., including 46 Enterobacter cloacae), and 71 non-fermenting bacteria (69 Pseudomonas spp., including 54 Pseudomonas aeruginosa, and 2 Stenotrophomonas maltophilia) being identified by the MALDI-TOF-MS method. Antibiotic susceptibility testing was performed by disk diffusion method and E-Test was used to determine MICs for imipenem. Several genes implicated in beta-lactams resistance were characterized by PCR and sequencing. Carbapenem resistance was detected among 12 isolates; nine E. coli (blaNDM-1 (n = 8); blaNDM-1 + blaVIM-2 (n = 1)) and three P. aeruginosa were carbapenem-resistant by loss of OprD porin. The whole-genome sequencing of P. aeruginosa 97H was determined using Illumina MiSeq sequencer, typed ST285, and harbored blaOXA-494. Other genes were also detected, notably blaTEM (n = 23), blaCTX-M-1 (n = 10) and blaCTX-M-9 (n = 6). These new epidemiological data imposed new surveillance strategies and strict hygiene rules to decrease the spread of multidrug-resistant bacteria in this area.

scholarly journals Molecular characterization of resistance mechanisms in Pseudomonas aeruginosa isolates resistant to carbapenems

2018 ◽  
Vol 11 (12) ◽  
pp. 935-943 ◽  
Author(s):  
Mona Shaaban ◽  
Ahmed Al-Qahtani ◽  
Mohammed Al-Ahdal ◽  
Rasha Barwa
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Real Time ◽  
Real Time Pcr ◽  
Treatment Options ◽  
Carbapenem Resistance ◽  
Pulse Field ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Pcr Analysis ◽  
Carbapenem Resistant

Introduction: Emergence of carbapenem resistance in Pseudomonas aeruginosa increases the therapeutic dilemma. In this study, we investigated various mechanisms involved in the resistance of P. aeruginosa clinical isolates to carbapenems. Methodology: P. aeruginosa isolates were isolated from different clinical samples. The antimicrobial susceptibility was evaluated by disc diffusion method. Carbapenemases were detected among carbapenem resistant isolates. Expression level of mexB and oprD was determined by real-time PCR. Molecular relatedness among isolates was detected based on pulse-field gel electrophoresis (PFGE). Results: Ninety P. aeruginosa isolates were purified from clinical specimens. High levels of resistance to imipenem and meropenem were detected in 16 isolates. PCR analysis of carbapenemases indicated the prevalence of Verona integron-encoded metallo-beta-lactamase (VIM); four isolates produced only VIM enzymes (VIM-1 or VIM-2), while the remaining twelve co-produced both VIM-1 or VIM-2 and NDM enzymes. Additionally, real-time PCR analysis elucidated high expression levels of mexB in seven of the carbapenem resistant isolates and low expression of oprD in seven isolates. The identified carbapenem-resistant isolates were clustered into eleven PFGE profiles where clusters E1 and E2 involved isolates exhibiting multiple carbapenemase genes (blaNDM-1, blaVIM-1 and blaVIM-2). Conclusion: Various mechanisms underlying carbapenem resistance have been detected in our P. aeruginosa cohort of isolates. Emergence of P. aeruginosa as a reservoir of multiple carbapenemases is increasing over time limiting the treatment options to this serious infection. This increases the urgency for infection control practices to reduce the incidence of this infection.

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