Characterisation of Plasmids in Multidrug Resistant Uropathogenic Gram-negative Bacterial Isolates

Introduction: Urinary Tract Infections (UTI) are one of the commonest conditions for which people seek medical care with an estimated 150 million episodes per annum worldwide. An unprecedented upsurge in the rate of development of antimicrobial resistance has reduced the therapeutic options leading to increased morbidity, prolonged hospital stays, development of complications. Majority of these infections are attributable to Gram negative bacteria which have now acquired resistance to almost all classes of antibiotics. Aim: To analyse the plasmid-mediated drug resistance and characterise the major plasmid families that are in circulation. Materials and Methods: A cross-sectional study comprising of a total of 95 non consecutive multidrug-resistant gram-negative bacterial isolates were subjected to Plasmid based replicon typing from January 2017 to June 2018. The 18 major replicons were divided in five multiplex and three uniplex Polymerase Chain Reaction (PCR) formats and the samples were subjected for plasmid characterisation and further sequencing of the plasmid Deoxyribonucleic Acid (DNA). The data obtained was analysed by Microsoft Excel software. Results: Escherichia coli, accounted for maximum n=51 (53.7%), Klebsiella pneumoniae n=19 (20%), Citrobacter sp n=11 (11.6%), miscellaneous gram negative n=14 (14.7%) The isolates exhibited a high degree of resistance to almost all tested antibiotics, sparing a few like Fosfomycin, Chloramphenicol, Imipenem, Amikacin. A total of 154 different plasmid families were detected from the 95 isolates. FIB replicon (24%), FIA (21%), F, W (20%), FIC, B/O (14%), Y (12%), I1 replicon (10.5%) were the major plasmid families detected in the present study. Conclusion: Many isolates exhibited the presence of more than one Incompatibility (Inc.) group plasmids, conferring multidrug resistance to the isolates. The study highlights the need for further research to study the association between plasmid families and their respective antibiotic resistance profiles for a given geographical niche and the need to devise further methods to target these epidemic plasmids.

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Plasmid Derived AmpC Genotypes among the Multidrug Resistant Enterobacteriaceae Strains Isolated from Urine Samples in Southern India

JOURNAL FOR CLINICAL AND DIAGNOSTIC RESEARCH ◽

10.7860/jcdr/2021/48194.14964 ◽

2021 ◽

Author(s):

P Ronni Mol ◽

Ganesan Shanthi ◽

Khalid Bindayna

Keyword(s):

Clinical Laboratory ◽

Multidrug Resistant ◽

Cross Sectional Study ◽

Urine Samples ◽

Pcr Analysis ◽

Genotype Distribution ◽

Cross Sectional ◽

Beta Lactamases ◽

Hospital Infection Control ◽

Tract Infections

Introduction: The most common pathogens causing Urinary Tract Infections (UTI) in community and hospital settings are Enterobacteriaceae. Antibiotic resistance is a major problem worldwide because of an increase in the use of antibiotics. Production of Extended Spectrum Beta-Lactamases (ESBLs) and AmpC beta-lactamases is the most common cause of resistance among Enterobacteriaceae (AmpC). Initially, AmpC β-lactamases received less attention globally, but now it has become a rising problem. Detection of AmpC β-lactamases expressing microbes is a requirement for addressing surveillance, for problems of hospital infection control, and for choosing optimal antimicrobial therapy. Aim: To study the genotype distribution of plasmid mediated AmpC β-lactamase produced in Enterobacteriaceaestrains isolated from urine samples. Materials and Methods: A cross-sectional study based on clinical laboratory surveillance was conducted from July 2019 to February 2020. Sixty Enterobacteriaceae isolates were identified by standard biochemical reactions. AmpC screening were done by cefoxitin disk diffusion and confirmed by an inhibitor-based assay using boronic acid. The presence of six plasmid mediated AmpC genes was determined by multiplex Polymerase Chain Reaction (PCR). Statistical Package for the Social Science (SPSS) version 20.0 was used to obtain descriptive data. Results: Among 60 Enterobacteriaceae isolates, 23 (38.3%) were cefoxitin-resistant isolates which contain Escherichia colistrain (n=17) while the remaining samples consist ofKlebsiella pneumoniae (n=5) and Proteus mirabilis strains (n=1). AmpC β-lactamase production was phenotypically confirmed in 12(20%) isolates and genotypically confirmed by PCR analysis in 16(26.6%) of all the urine isolates. In the present study, 3(13%), 2 (8.6%) of cefoxitin resistant isolates harboured the DHA, EBC gene and 1(4.3%) each harboured FOX and CIT gene, and 9(39.1%) harboured a combination of the genes. Conclusion: The present study suggested the predominant existence of plasmid mediated AmpC producers in Multi-Drug Resistant (MDR) Escherichia coli and Klebsiella pneumoniae. We suggest continuous surveillance is important to effectively control the spread of these strains and for optimal clinical outcome.

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Multidrug-resistant urinary tract isolates of escherichia coli: Frequency and patient demographics in a Tertiary Care Hospital.

The Professional Medical Journal ◽

10.29309/tpmj/2020.27.02.3854 ◽

2020 ◽

Vol 27 (02) ◽

pp. 335-340

Author(s):

Salman Azhar ◽

Muhmmad Wasif Baig ◽

Shahid Rasool ◽

Rizwan Rasool Khan ◽

Talha Munir ◽

...

Keyword(s):

Urinary Tract ◽

Tertiary Care Hospital ◽

Tertiary Care ◽

Multidrug Resistant ◽

Cross Sectional Study ◽

Care Hospital ◽

Cross Sectional ◽

E Coli ◽

Patient Demographics ◽

Tract Infections

Urinary tract infections (UTIs) are a major burden to the health care as it is estimated that around 150 million UTIs occur yearly worldwide. Enterobacteriaceae are the most common agent causing serious urinary tract infections; and MDR cases are increasing day by day. Objectives: To determine the frequency and patient demographics of multidrug resistant urinary tract isolates of Escherichia Coli in a Tertiary Care Hospital. Study Design: Retrospective cross sectional study. Setting: Medicine Department of Madinah Teaching Hospital / The University of Faisalabad, Faisalabad. Period: From May 2016 to Sep 2018. Material & Methods: 187 patients of age 15 to 90 years with positive E. coli on urine culture and sensitivity were included in this Retrospective cross sectional study. All those patients with history of dysuria (pain during urination) or frequent urination (more than 7 times per day) were advised urine complete analysis and those with >5 WBCs or pus cells /HPF or having positive for leukocyte esterase and/or nitrite, were advised urine culture and sensitivity. Main outcome variable was the frequency of MDRE cases among the culture positive E. coli UTI patients. Results: Among all the cases of E. coli UTI, frequency of MDRE UTI was 66.8% (n=125) and rest 33.2% (n= 62) cases were not MDR UTI. 97.3% patients were resistant to lactam antibiotics, 95.7% were resistant to quinolones and 68.4% were resistant to aminoglycosides.

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Fluoroquinolones Resistance Among Bacterial Uropathogens In A Tertiary Care Hospital Of Western Nepal

Birat Journal of Health Sciences ◽

10.3126/bjhs.v5i3.33682 ◽

2020 ◽

Vol 5 (3) ◽

pp. 1137-1141

Author(s):

Dharm Raj Bhatta ◽

Rajani Shrestha ◽

Deependra Hamal ◽

Supram HS ◽

Niranjan Nayak

Keyword(s):

Tertiary Care ◽

Cross Sectional Study ◽

Empirical Treatment ◽

Resistance Pattern ◽

Care Hospital ◽

Klebsiella Species ◽

Cross Sectional ◽

Gram Negative ◽

Microbiological Techniques ◽

Tract Infections

Introduction: Urinary tract infections are one of the most common clinical illnesses in developing countries. Most of the infections are treated empirically due to unavailability of laboratory facilities. Fluoroquinolones are frequently used for empirical treatment of UTIs. Objectives: This study was aimed to determine prevalence and antibiotic resistance pattern of bacterial uropathogens, especially to fluoroquinolones. Methodology: This hospital based cross sectional study was carried out at Manipal Teaching Hospital, Pokhara, Nepal. A total of 2450 mid stream urine samples were included in this study. Isolation, identification and antibiotic susceptibility testing of the isolates was performed by standard microbiological techniques. Results: Significant bacteriuria was detected in 258 (10.5%), insignificant bacteruria in 614 (25%), growth of multiple organisms in 238 (9.7%) while no growth was recorded in 1340 (54.7%) samples. E coli was the commonest isolate followed by Klebsiella species. Fluoroquinolones resistance among Gram negative isolates was high. Most of the Gram negative isolates were susceptible to amikacin, gentamicin, nitrofurantoin and imipenem. Conclusion: High resistance to fluoroquinolones, cotrimoxazole and cephalosporins dictates these to be unsuitable for empirical treatment of UTIs. Amikacin, gentamicin, nitrofurantoin and imipenem were the most effective antibacterials.

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Clinico-Microbiological Investigation on Fosfomycin and Tigecycline Resistant Gram-Negative Bacilli Isolated from Urinary Tract Infections: A Potential Resurgence

Jundishapur Journal of Microbiology ◽

10.5812/jjm.99990 ◽

2020 ◽

Vol 13 (7) ◽

Author(s):

Maryam Dastborhan ◽

Mojtaba Varshochi ◽

Alka Hasani ◽

Leila Dehghani ◽

Elghar Soltani ◽

...

Keyword(s):

Urinary Tract ◽

Urinary Tract Infections ◽

Cross Sectional Study ◽

Cross Sectional ◽

In Vitro Susceptibility ◽

Gram Negative ◽

The Difference ◽

Conventional Antibiotics ◽

Tract Infections

Background: Traditional antibiotics are no longer as effective as before for controlling pathogens associated with urinary tract infections (UTI), which shows the necessity of developing new and more effective antibiotics. Objectives: The current study aimed to evaluate in vitro susceptibility of fosfomycin and tigecycline towards common antibiotic-resistant Gram-negative bacilli isolated from the urinary tract. Besides, clinico-microbiological on fosfomycin and tigecycline resistant Gram-negative bacilli was investigated. Methods: In this descriptive cross-sectional study, 150 resistant Gram-negative bacilli were isolated from urine specimens send for culture, and antibiotic susceptibility assessment to the Division of Microbiology of Sina Hospital affiliated to Tabriz University of Medical Sciences which were collected from April-September 2017 are included. Antibiotic susceptibilities were evaluated according to the Clinical and Laboratory Standards published by the Institute and the criteria of the Food and Drug Administration. Results: Of 150 isolates, 138 (92%) were susceptible, and 2 (1.3%) were resistant to both fosfomycin and tigecycline, as confirmed by disk diffusion and Epsilonmeter tests. The difference was statistically significant (P = 0.001). Conclusions: Based on the results, resistance to the conventional antibiotics prescribed for the treatment of UTI was significantly high. Fosfomycin and tigecycline have an appropriate antimicrobial activity towards Gram-negative-resistant isolates involved in UTIs.

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Phenotypic and genotypic detection of carbapenemase enzymes producing gram-negative bacilli isolated from patients in Khartoum State

F1000Research ◽

10.12688/f1000research.12432.1 ◽

2017 ◽

Vol 6 ◽

pp. 1656

Author(s):

R.A. Dahab ◽

Alamin Mohamed Ibrahim ◽

Hisham N. Altayb

Keyword(s):

Treatment Options ◽

Multidrug Resistant ◽

Cross Sectional Study ◽

Biochemical Tests ◽

Isolation And Identification ◽

Cross Sectional ◽

Gram Negative ◽

Clinical Specimens ◽

Carbapenem Resistant ◽

Life Threatening

Background: Carbapenems are used as antibiotics of last resort for treating infections due to multidrug-resistant Gram-negative bacilli, but emergence of Carbapenem resistant Gram-negative bacilli have been reported due to the production of Carbapenemase enzymes that significantly limits treatment options for life-threatening infections. Objective: This study aimed to detect Carbapenem resistant Gram-negative bacilli from patients attended to different hospitals in Khartoum state and to detect Carbapenemase enzymes production by phenotypic and genotypic methods. Methods: A hospital based cross sectional study was conducted in Khartoum state in the period from February to August 2016. Hundred and forty nine Gram-negative bacilli bacteria were isolated from different clinical specimens. Blood agar, Chromogenic agar media, MacConkey agar, XLD mediaandstandard biochemical tests were used for isolation and identification of Gram-negative bacilli from different samples. Standard antimicrobial susceptibility testing to Carbapenem antibiotic was performed for all isolates, then detection of Carbapenemase enzymes production for the resistant isolates was performed using Modified Hodge Test and PCR. Results: Hundred and forty nine Gram-negative bacilli were isolated from 147 different clinical specimens. The most predominant Gram-negative bacilli isolates was E.coli (54.4%), followed by Klebsiella species (29.5%). More than fifty percent of the isolates were Carbapenem resistant. Fifty six percent of the resistant isolates were positive by Modified Hodge Test. By using PCR, 17.3% of resistant organisms were harbored blaOXA48 gene, and 6.7% harbored blaIMP gene. E.coli was the most bacteria that harbored the blaoxa48 followed by Klebsiella species. blaIMP gene was harbored only by E.coli. Conclusion: The percentage of resistance to Carbapenems due to production of Carbapenemase enzymes is very high in Sudan.BlaOXA48 gene is more predominant than blaIMP in this study.

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A Cross -Sectional Study of Bacteriological Profile of Lower Respiratory Tract Infections in Neurosurgery Intensive Care Unit in Central Kerala

Journal of Evidence Based Medicine and Healthcare ◽

10.18410/jebmh/2021/389 ◽

2021 ◽

Vol 8 (24) ◽

pp. 2077-2082

Author(s):

Satheesh Chandra Sugatha Rao ◽

Shara Beena

Keyword(s):

Intensive Care ◽

Respiratory Tract ◽

Lower Respiratory Tract ◽

Respiratory Tract Infections ◽

Cross Sectional Study ◽

Drug Resistant ◽

Bacterial Isolates ◽

Lower Respiratory Tract Infections ◽

Cross Sectional ◽

Tract Infections

BACKGROUND Lower respiratory tract infections are the most common bacterial infections in neurosurgery intensive care units (NSICU), resulting in high overall mortality. The emergence of antibiotic resistant pathogens poses a challenge to their empiric treatment. Regular surveillance of the prevalent strains and their susceptibility pattern, helps to revise the antibiotic policies and aids in better management of the patient. METHODS A cross sectional study was conducted in the Department of Neurosurgery and Microbiology, Government Medical College, Thrissur, over a period of 1 year, using lower respiratory tract specimens of 190 patients with acute respiratory symptoms admitted in neurosurgery intensive care unit. The specimens collected aseptically were processed immediately. Following culture, the bacterial isolates were identified using standard methods and antibiotic susceptibility was done by Kirby Bauer disc diffusion method. The data obtained was coded and entered in Microsoft Excel and expressed as percentage. RESULTS Bacterial isolates were obtained from 74 % samples. 82 % isolates were monomicrobial and 18 % were polymicrobial. Majority of the isolates were gram negative bacteria (94 %) followed by gram positive bacteria (6 %). The common gram-negative isolates were K. pneumoniae (36 %), A. baumannii (29 %), P. aeruginosa (20 %). Methicillin-resistant staph (MRSA) (3 %) and methicillinsusceptible staph (MSSA) (3 %) accounted for the gram positive cocci. 61 % isolates were multi drug resistant (MDR). Most common MDR organism was A. baumannii. It was observed that 26 % isolates were extended spectrum betalactamase (ESBL) producers. A high rate of resistance to cephalosporins, beta lactams, fluoroquinolones, aminoglycosides, and cotrimoxazole was observed. An emerging resistance to carbapenems was observed. CONCLUSIONS Proper microbiological work up and antibiotic stewardship programmes can limit spread of resistant organisms, thereby reducing the medical and economic burden of the patient. KEYWORDS Lower Respiratory Tract Infection, Neurosurgery ICU, Antimicrobial Susceptibility Tests, Multi Drug Resistant Organisms

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High prevalence of multidrug-resistant non-fermentative Gram-negative bacilli harboring bla and bla metallo-beta-lactamase genes in Birjand, south-east Iran

Iranian Journal of Microbiology ◽

10.18502/ijm.v13i4.6971 ◽

2021 ◽

Author(s):

Mohammad Hasan Namaei ◽

Masoud Yousefi ◽

Parvin Askari ◽

Babak Roshanravan ◽

Ali Hashemi ◽

...

Keyword(s):

Antibiotic Resistance ◽

Multidrug Resistant ◽

Cross Sectional Study ◽

Disk Diffusion ◽

Molecular Characteristics ◽

Beta Lactamase ◽

Cross Sectional ◽

Disk Diffusion Test ◽

Gram Negative ◽

Carbapenem Resistant

Background and Objectives: Non-fermentative Gram-negative Bacilli (NFGNB) is known as a major cause of health- care-associated infections with high levels of antibiotic resistance. The aim of this study was to investigate the antibiotic resistance profiles and molecular characteristics of metallo-beta-lactamase (MBL)-producing NFGNB. Materials and Methods: In this cross-sectional study, the antibiotic resistance profile of 122 clinical NFGNB isolates was determined by the Kirby-Bauer disk diffusion and microdilution broth methods. Bacterial isolates were investigated for the detection of MBLs production using the combination disk diffusion Test (CDDT). The existence of bla , bla , and bla NDM genes in all carbapenem-resistant isolates was determined employing polymerase chain reaction (PCR) assays. Results: High resistance in Pseudomonas aeruginosa was reported to cefotaxime and minocycline, whereas Acinetobacter baumannii isolates were highly resistant to all antibiotics except colistin. Multidrug resistance (MDR)-NFGNB (66% vs. 12.5%, P=0.0004) and extensively drug resistant (XDR)-NFGNB (55.7% vs. 12.5%, P=0.001) isolates were significantly more common in hospitalized patients than in outpatients. The production of MBL was seen in 40% of P. aeruginosa and 93.3% of A. baumannii isolates. It was found that 33.3% and 46.7% of carbapenem-resistant P. aeruginosa isolates, and 13.3% and 28.9% of carbapenem-resistant A. baumannii isolates were harboring bla IMP-1 and bla VIM-1 genes, respectively. The incidence of MDR (98.2% vs. 28.3%, P<0.001) and XDR (96.4% vs. 11.7%, P<0.001) in MBL-producing NFGNB isolates was significantly higher than non-MBL-producing isolates. Conclusion: This study demonstrated a higher rate of resistance among NFGNB isolates with an additional burden of MBL production within them, warranting a need for robust microbiological surveillance and accurate detection of MBL producers among the NFGNB.

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Bacteremia in Diarrheal Children With Severe Pneumonia

Global Pediatric Health ◽

10.1177/2333794x19862462 ◽

2019 ◽

Vol 6 ◽

pp. 2333794X1986246

Author(s):

Haimanti Saha ◽

Lubaba Shahrin ◽

Monira Sarmin ◽

Tahmeed Ahmed ◽

Mohammod Jobayer Chisti

Keyword(s):

Diarrheal Disease ◽

Severe Pneumonia ◽

Cross Sectional Study ◽

Childhood Mortality ◽

Gram Negative Bacteria ◽

Limited Information ◽

Bacterial Isolates ◽

Cross Sectional ◽

Gram Negative ◽

The Difference

Objectives. Diarrhea and pneumonia are the leading causes of under-5 childhood mortality. However, there is limited information on bacterial etiology of severe pneumonia in children with diarrhea. We analyzed bacterial pathogens from the blood of children under the age of 5 years. Methods. In this retrospective cross-sectional study, we studied all children having severe pneumonia with or without diarrhea admitted to the icddr,b (International Centre for Diarrheal Disease Research, Bangladesh) who had their blood culture done during January 2014 to December 2014. Results. Among a total of 159 study children, 118 had diarrhea. There were 13 bacterial isolates, and predominant organisms were gram-negative bacteria (11/13, 85%). Children with diarrhea coexisting with severe pneumonia proportionately had higher bacteremia (12/141 [10.16%] vs 1/41 [2.43%]), but the difference was not statistically insignificant ( P = .186). Conclusion. We recognized that the coexistence of diarrhea and severe pneumonia had proportionately higher bacteremia, especially gram-negative bacteria compared with those without diarrhea. The results emphasize the trend of bacterial etiology of pneumonia in children with diarrhea and may warrant revised antibiotics guideline for their management.

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Extended Spectrum Beta-lactamase Producing Gram Negative Bacterial Isolates from Urine of Patients Visiting Everest Hospital, Kathmandu, Nepal

Tribhuvan University Journal of Microbiology ◽

10.3126/tujm.v6i0.26575 ◽

2019 ◽

Vol 6 ◽

pp. 26-31 ◽

Cited By ~ 1

Author(s):

Nisha Guragain ◽

Aashish Pradhan ◽

Binod Dhungel ◽

Megha Raj Banjara ◽

Komal Raj Rijal ◽

...

Keyword(s):

Bacterial Species ◽

Cross Sectional Study ◽

Urine Samples ◽

Diffusion Method ◽

Beta Lactamase ◽

Bacterial Isolates ◽

Cross Sectional ◽

Gram Negative ◽

Extended Spectrum Beta Lactamase ◽

Extended Spectrum

Objectives: The study was aimed to determine the prevalence of Extended Spectrum Beta Lactamase (ESBL) producing Gram negative pathogens from urine samples along with their antimicrobial resistance. Methods: This cross-sectional study was conducted from December 2015 to May 2016 at Everest Hospital, Kathmandu. Mid-stream urine samples were collected and processed for culture by standard loop streak method. Identified bacterial isolates were tested for Antibiotic Susceptibility by modified Kirby Bauer disc diffusion method and, were subjected to ESBL screening by using 30µg cefotaxime and ceftazidime. ESBL production was confirmed by combination disc method. Results: Of the three hundred urine samples, 22.7% (67/300) showed significant growth. Four different bacterial species were identified. Among the isolates, E. coli was the most common pathogen (71.64%) followed by Klebsiella pneumoniae (14.92%), Pseudomonas spp (8.95%) and Acinetobacter spp (4.48%). Altogether 92.54% (n=62) isolates were sensitive to gentamicin, 89.55% (n=60) to amikacin, and 79.10% (n=53) to nitrofurantoin. 70.10% (n=47) isolates were resistant to antibiotic ampicillin while 62.68% (n=42) were found as Multi-Drug Resistant (MDR) and 29.8% (n=20) were ESBL producers. Conclusions: The overall prevalence of MDR and ESBL among uropathogens is low in comparison to other studies though it is essential to have a regular monitoring of ESBL producing clinical isolates in laboratory practice.

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Prevalence of Ciprofloxacin Resistance among Gram-Negative Bacilli Isolated From Different Specimens at a General Hospital in Riyadh, Saudi Arabia

Journal of Shaheed Suhrawardy Medical College ◽

10.3329/jssmc.v5i1.16197 ◽

2013 ◽

Vol 5 (1) ◽

pp. 7-10

Author(s):

Mohammad Afzal Hossain ◽

Shirin Mohal ◽

Md Abdullah Yusuf ◽

Md Saiful Islam ◽

Md Rafiqul Islam ◽

...

Keyword(s):

Saudi Arabia ◽

General Hospital ◽

Inhibitory Concentration ◽

Bacterial Species ◽

Cross Sectional Study ◽

Test Results ◽

Cross Sectional ◽

Gram Negative ◽

Ciprofloxacin Resistance ◽

Tract Infections

Background: Resistance among the different classes of antimicrobials including fluoroquinolones has arisen in a multitude of bacterial species in the hospitals and in the community. Objective: The purpose of the present study was to measure the prevalence of ciprofloxacin resistance as well as to evaluate the susceptibility pattern of the isolates to various antibiotics. Methodology: This cross sectional study was carried out in the Department of Microbiology at a General Hospital in Riyadh, Saudi Arabia from July'2009 to December'2009. Different clinical specimens were collected and culture was done in the appropriate media. Antibiotic susceptibility testing was performed. Gram negative bacterial isolates with decreased susceptibility to ciprofloxacin were tested for minimum inhibitory concentration (MIC) by using the E-test. Results: A total of 1246 gram-negative bacilli (GNB) were isolated from different specimens of which 486(39.0%) isolates were resistant to ciprofloxacin. The MIC of these isolates ranged from 4 to 32 ?g/ml. Most of the Ciprofloxacin resistant isolates were from urinary tract infections. Conclusion: The prevalence of ciprofloxacin resistance in different specimens is high. DOI: http://dx.doi.org/10.3329/jssmc.v5i1.16197 J Shaheed Suhrawardy Med Coll, 2013;5(1):7-10

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