Distribution of Iron Uptake Systems Encoding Genes Among the Clinical Isolates of Escherichia coli Compared to Foodstuffs Isolates

2020 ◽  
Vol 20 (4) ◽  
pp. 517-522
Author(s):  
Hassan Mahmoudi ◽  
Hadi Hossainpour ◽  
Mohammad Moradi ◽  
Mohammad Yousef Alikhani
Keyword(s):  
Escherichia Coli ◽  
Iron Uptake ◽  
Food Poisoning ◽  
Food Samples ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
E Coli ◽  
Microbiological Methods ◽  
Polymerase Chain ◽  
Encoding Genes

Background: Bacteria require iron ions to grow and infect the host, which, by using iron uptake systems, acquire free iron from their host cell. Escherichia coli is one of the most important pathogens to cause food poisoning and clinical infections. The aim of this study was to assess the distribution of iron uptake systems encoding genes in clinical isolates of E.coli compared to food samples isolates. Materials and Methods: This investigation was conducted to determine the prevalence of E. coli isolated from various sources of food and clinical specimens. The E. coli isolates confirmed by the standard microbiological methods. The isolates were examined for the presence of iut A and iuc A genes by specific primers using the polymerase chain reaction technique. Results: A total of 100 and 50 isolates of E. coli were collected from clinical samples and foodstuffs, respectively. The prevalence of E. coli in the food and clinical samples was 33.33% and 64.10%, respectively. The frequency of iut A and iuc A genes in the food and clinical isolates were 76%-84% and 86% - 83%, respectively. Conclusion: Our results showed that the prevalence of E. coli isolates with iut A and iuc A genes was relatively higher compared to many previous studies. The existence of these genes in E. coli strains is likely to be related to pathogenicity in those strains, which requires further studies in the future.

scholarly journals IDENTIFICATION OF AMPC Β-LACTAMASE-PRODUCING CLINICAL ISOLATES OF ESCHERICHIA COLI

2017 ◽  
Vol 10 (12) ◽  
pp. 357
Author(s):  
Tanushree Barua Gupta ◽  
Malini Shariff ◽  
Thukral Ss ◽  
S.s Thukral
Keyword(s):  
Escherichia Coli ◽  
Detection Method ◽  
Clinical Isolates ◽  
Confirmatory Test ◽  
Chain Reaction ◽  
E Coli ◽  
Resistance To Penicillin ◽  
Polymerase Chain ◽  
Third Generation Cephalosporins

  Objective: Indiscriminate use of β-lactam antibiotics has resulted in the emergence of β-lactamase enzymes. AmpC β-lactamases, in particular, confer resistance to penicillin, first-, second-, and third-generation cephalosporins as well as monobactams and are responsible for antibiotic resistance in nosocomial pathogens. Therefore, this study was undertaken to screen nosocomial Escherichia coli isolates for the presence and characterization of AmpC β-lactamases. The study also envisaged on the detection of inducible AmpC β-lactamases and extended-spectrum β-lactamases (ESBLs) in AmpC β-lactamase-producing E. coli.Methods: A total of 102 clinical isolates of E. coli, were subjected to cefoxitin screening, and screen-positive isolates were further subjected to inhibitor-based detection method, phenotypic confirmatory test, disc antagonism test, polymerase chain reaction (PCR), and isoelectric focusing (IEF).Results: In this study, 33% of E. coli were resistant to cefoxitin, of which 35% were found to be positive for AmpC β-lactamase by inhibitor-based phenotypic test. Of the AmpC-positive isolates, 83% were positive for ESBLs, whereas 25% were producing inducible AmpC β-lactamases. PCR and IEF showed CIT and EBC types of AmpC β-lactamases present in the tested isolates.Conclusion: Our study showed the presence of inducible AmpC enzymes and ESBLs in E. coli isolates and PCR identified more isolates to be AmpC producers.

scholarly journals Molecular Investigation of hemolytic phospholipase C Encoding Genes in Clinical Isolates of Pseudomonas aeruginosa

2018 ◽  
Vol 9 (SPL1) ◽  
Author(s):  
Rasha Hadi Saleh ◽  
Habeeb S Naher ◽  
Mohammed AK Al-saadi
Keyword(s):  
Phospholipase C ◽  
Virulence Genes ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
Burn Wound ◽  
Molecular Investigation ◽  
Genes Encoding ◽  
Stool Samples ◽  
Polymerase Chain ◽  
Encoding Genes

This study is aimed to isolate P.aeruginosa from different clinical cases and to detect the prevalence of virulence genes encoding hemolytic phospholipase C(plcH)in these clinical isolates. In this study a total of 422 clinical samples including burn,wound,ear,urine,abscess and stool were aseptically taken from out- and inpatients who admitted into two hospitals in Hilla City (Teaching Al-Hilla Hospital and Babylon Hospital for Maternity and children during a period of three months. All samples were subjected to bacterial cultivation for the isolation of P.aeruginosa. The isolated P.aeruginosa was diagnosed depended on morphological,biochemical and molecular standard characteristics. Hemolytic phospholipase Cencoding genes(plcH) were detected by PCR and the amplification products were separated in 1% agarose gels containing ethidium bromide. Out of 422 samples,P.aeruginosa was isolated from 54 samples (12.8%). The distribution of these isolates were: 22 (55%) from burn samples,2; (50%) from diabitics foot samples,8 (14.8%) from wound samples, 8 (32%) from ear samples,3 (11%) from abscess samples, 7 (4%) from stool samples,4 (4%) from urine samples and 0 sputum samples. The genotypic properties of hemolytic phospholipase C (plcH )toxins was detected by polymerase chain reaction (PCR). The results of this study revealed that(plcH )gene found in 13/20 (65%)of isolates.

scholarly journals Modified CIM-Test As a Useful Tool to Detect Carbapenemase Activity Among Extensively Drug- Resistant Klebsiella Pneumoniae, Escherichia Coli and Acinetobacter Baumannii

2021 ◽  
Author(s):  
Abed Zahedi bialvaei ◽  
Alireza Dolatyar Dehkharghani ◽  
Farhad Asgari ◽  
Firouzeh Shamloo ◽  
Parisa Eslami ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Klebsiella Pneumoniae ◽  
Acinetobacter Baumannii ◽  
Sensitivity And Specificity ◽  
E Coli ◽  
Carbapenem Resistant ◽  
Polymerase Chain ◽  
Phenotypic Methods ◽  
Encoding Genes ◽  
Positive Results

Abstract Background Timely detection of carbapenemases is essential for developing strategies to control the spread of infections by carbapenem-resistant isolates. The purpose of our study was to determine the epidemiology of carbapenemase genes among carbapenem resistant isolates of Acinetobacter baumannii, Klebsiella pneumoniae and Escherichia coli and to compare efficacy of modified Hodge Test (MHT), Carba NP and modified carbapenem inactivation method (mCIM) tests. Methods A total of 122 carbapenem-resistant clinical isolates including 77 K. pneumoniae, 39 A. baumannii, and six E. coli were collected from hospitalized patients. Three phenotypic methods, including MHT, Carba NP test and mCIM were used for investigation of carbapenemase production. In addition, polymerase chain reaction (PCR) was performed to detect carbapenemase encoding genes. Results The sensitivity and specificity of the MHT were 75.0% and 100% respectively. In addition, CarbaNP displayed 80.8% sensitivity and 100% specificity, whereas the sensitivity and specificity were 90.4% and 100% for the mCIM test, respectively. Among carbapenem-resistant isolates, 70, 84 and 87 isolates exhibited positive results according to MHT, CarbaNP test and mCIM, respectively. PCR indicated the presence of one or more carbapenemase genes in 119 of carbapenem-resistant isolates, with blaKPC and blaVIM being the most commonly encountered. Co-production of ‘KPC and VIM’, ‘KPC and IMP’ and ‘KPC and OXA-48’ was detected in nine, seven and three isolates, respectively. Conclusion Our results confirm that the mCIM test is a useful tool for the reliable detection of carbapenemases-activity in enterobacterial isolates, especially in clinical microbiological laboratories with limited resources.

scholarly journals Virulence factors, antimicrobial resistance, and plasmid content of Escherichia coli isolated in swine commercial farms

2010 ◽  
Vol 62 (1) ◽  
pp. 30-36 ◽  
Author(s):  
M.M. Costa ◽  
G. Drescher ◽  
F Maboni ◽  
S.S. Weber ◽  
A. Schrank ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Resistance ◽  
Virulence Factors ◽  
Intestinal Content ◽  
Specific Pattern ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
Plasmid Content ◽  
E Coli ◽  
Resistance Patterns

Virulence factors and antimicrobial resistance patterns of Escherichia coli isolates were evaluated. A total of 80 E. coli isolates were evaluated, being 64 from clinical samples (intestinal content and fragments of organs from diarrheic piglets), seven from feces of clinically healthy piglets and sows, and nine environmental samples (five from facilities, two from feed, one from insect, and one from waste). Molecular characterization was performed by PCR detection of fimbriae and toxin genes and plasmid content determination. The isolates were also characterized according to their resistance or sensitivity to the following drugs: ampicillin, trimethoprim:sulfamethoxazole, tetracycline, amikacine, colistin, norfloxacin, florfenicol, enrofloxacin, cefalexin, trimethoprim, neomycin, chloramphenicol, and gentamicin. From 80 E. coli isolates, 53.8% were classified as enterotoxigenic E. coli (ETEC), 2.5% were shiga toxin-producing E. coli (STEC), and 43.8% showed a non specific pattern and were unclassified. One fecal isolate from non-diarrheic piglet was classified as ETEC by PCR. Clinical isolates showed resistance mainly for tetracycline and trimethoprim:sulfamethoxazole. Plasmidial DNA was observed in 70 isolates, being 78.5% of clinical isolates, 8.57% of non-diarrheic feces, and 12.8% of environment.

scholarly journals Modified CIM test as a useful tool to detect carbapenemase activity among extensively drug-resistant Klebsiella pneumoniae, Escherichia coli and Acinetobacter baumannii

2021 ◽  
Vol 71 (1) ◽  
Author(s):  
Abed Zahedi Bialvaei ◽  
Alireza Dolatyar Dehkharghani ◽  
Farhad Asgari ◽  
Firouzeh Shamloo ◽  
Parisa Eslami ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Klebsiella Pneumoniae ◽  
Acinetobacter Baumannii ◽  
Sensitivity And Specificity ◽  
E Coli ◽  
Carbapenem Resistant ◽  
Polymerase Chain ◽  
Phenotypic Methods ◽  
Encoding Genes ◽  
Positive Results

Abstract Purpose Timely detection of carbapenemases is essential for developing strategies to control the spread of infections by carbapenem-resistant isolates. The purpose of this study was to determine the epidemiology of carbapenemase genes among carbapenem-resistant isolates of Acinetobacter baumannii, Klebsiella pneumoniae, and Escherichia coli. In addition, the efficacy of the modified Hodge test (MHT), Carba NP test, and modified carbapenem inactivation method (mCIM) were compared. Methods A total of 122 carbapenem-resistant clinical isolates including 77 K. pneumoniae, 39 A. baumannii, and six E. coli were collected from hospitalized patients. Three phenotypic methods, including the MHT, Carba NP test, and mCIM were used for investigation of carbapenemase production. In addition, polymerase chain reaction (PCR) was performed to detect carbapenemase-encoding genes. Result The sensitivity and specificity of the MHT were 75.0% and 100%, respectively. In addition, Carba NP displayed 80.8% sensitivity and 100% specificity, whereas the sensitivity and specificity were 90.4% and 100% for the mCIM test, respectively. Among carbapenem-resistant isolates, 70, 84, and 87 isolates exhibited positive results according to the MHT, Carba NP test, and mCIM, respectively. PCR indicated the presence of one or more carbapenemase genes in 119 of carbapenem-resistant isolates, with blaKPC and blaVIM being the most commonly encountered. Co-production of ‘KPC and OXA-48’, ‘KPC and VIM’, and ‘KPC and IMP’ was detected in three, nine, and seven isolates, respectively. Conclusion Our results confirm that the mCIM test is a useful tool for the reliable detection of carbapenemase activity in enterobacterial isolates, especially in clinical microbiological laboratories with limited resources.

scholarly journals Phenotypic Detection of Extended-Spectrum ß-lactamase Produced by Escherichia coli Isolates from Health facilities in Otukpo, Benue State Nigeria

2021 ◽  
Vol 2 (2) ◽  
pp. 7-13
Author(s):  
PS Utulo ◽  
EU Umeh ◽  
GM Gberikon ◽  
PO Abba
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Agents ◽  
Health Facilities ◽  
Molecular Techniques ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Esbl Production ◽  
E Coli ◽  
Extended Spectrum ◽  
Microbiological Methods

Beta-lactamase production by Escherichia coli and other bacteria is one of the most important mechanism of resistance to beta-lactam antibiotics such as Penicillins and Cephalosporins which are the most commonly used antibiotics in the treatment of bacterial infections in Hospitals. Resistance to antibiotics is emerging worldwide as a threat to human health resulting in limitations of therapeutic options. Susceptibility tests of microorganisms to antimicrobial agents do not reveal production of Extended-Spectrum ß-lactamase, hence the need to detect their presence by phenotypic or molecular techniques. This study is aimed at determining the prevalence of ESBL-production by Escherichia coli isolates from clinical samples collected from some selected health facilities in Otukpo, using phenotypic detection. Four hundred (400) specimens were collected from four different health facilities in Otukpo. Specimens collected included urine, blood, stool and wound swabs from 222 females and 178 males. Standard microbiological methods were employed for isolation, identification and characterization of Escherichia coli isolates from these specimens. Antibiotic susceptibility of the E. coli isolates was determined using Kirby-Bauer disc diffusion method while double disc synergy test (DDST) was used for ESBL production. Sixty-eight, (17%) of the 400 specimens tested for ESBL production were positive. Isolates from stool 22(22.7%) had the highest prevalence of ESBL-producing E. coli, followed by isolates from blood 4(21.4%). Urine isolates had the least prevalence 31(13.8%). Isolates from female subjects, 38(17.1%) had higher positivity rate than their male counterpart 30(16.9%). Otukpo General Hospital had the highest prevalence 20(20.0%) of ESBL followed by Otia Hospital 18(18.0%). Comprehensive health center, Otukpo had the least prevalence 14(14.0%). There is a high prevalence of ESBL production by Escherichia coli isolated from health facilities in Otukpo. Routine drug resistance surveillance therefore has become necessary to guide the appropriate and judicious antibiotic use.

scholarly journals Detection of CTX-M-type ESBLs from Escherichia coli Clinical Isolates from a Tertiary Hospital, Malaysia

2019 ◽  
Vol 16 (3(Suppl.)) ◽  
pp. 0682 ◽  
Author(s):  
MKK Et al.
Keyword(s):  
Escherichia Coli ◽  
Clinical Specimen ◽  
Pcr Amplification ◽  
Tertiary Hospital ◽  
Clinical Isolates ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Disk Diffusion Method ◽  
E Coli ◽  
Resistance Rates

The present study aims to detect CTX-M-type ESBL from Escherichia coli clinical isolates and to analyze their antibotic susceptibility patterns. One hundred of E. coli isolates were collected from different clinical samples from a tertiary hospital. ESBL positivity was determined by the disk diffusion method. PCR used for amplification of CTX-M-type ESBL produced by E. coli. Out of 100 E. coli isolates, twenty-four isolates (24%) were ESBL-producers. E. coli isolated from pus was the most frequent clinical specimen that produced ESBL (41.66%) followed by urine (34.21%), respiratory (22.23%), and blood (19.05%).  After PCR amplification of these 24 isolates, 10 (41.66%) isolates were found to possess CTX-M genes. The CTX-M type ESBL producing E. coli against antibiotics belonging to different families showed the highest resistance rates to Ampicillin (100%), Cefotaxime (97%), Cefuroxime (95%), and Ciprofoxacin (86%). Carbapenem groups of antibiotics, Meropenem (89%) and Imipenem (85%) have the highest susceptibility rate among all antibiotics used in this study. The outcome of the antimicrobial susceptibility testing of significant CTX-M- type ESBL producing E. coli could be useful to avoid failure or prolong treatments.

scholarly journals The prevalence and mechanism of triclosan resistance in Escherichia coli isolates from urine samples in Wenzhou, China

2020 ◽  
Author(s):  
Weiliang Zeng ◽  
Wenya Xu ◽  
Ye Xu ◽  
Wenli Liao ◽  
Yajie Zhao ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Efflux Pump ◽  
Cross Resistance ◽  
Chain Reaction ◽  
E Coli ◽  
Resistant Strains ◽  
Polymerase Chain ◽  
Triclosan Resistance ◽  
Encoding Genes

Abstract Background Widespread use of triclosan has been reported to cause its residue in urine, which provides an environment of long-term exposure to triclosan for intestinal Escherichia coli. We aimed to determine the triclosan and antibiotic resistance characteristics of Escherichia coli strains isolated from urine, and further investigate the resistance mechanism and molecular epidemic characteristics of triclosan resistant Escherichia coli isolates. Methods A total of 200 non-repetitive E. coli strains from urine samples were obtained and identified. The minimum inhibitory concentrations (MICs) of triclosan and antibiotics, fabI mutation, efflux pump activity, expression of 14 efflux pump encoding genes and epidemiological characteristics were detected with agar dilution method, polymerase chain reaction (PCR), carbonyl cyanide 3-chlorophenylhydrazone (CCCP) inhibition test, quantitative real-time polymerase chain reaction (RT-qPCR), multilocus sequence typing (MLST) and pulse field gel electrophoresis (PFGE) in all triclosan resistant isolates. Furthermore, we also investigated the effect of triclosan exposure in vitro on resistance in susceptible strains by serial passage experiment. Results Of 200 E. coli isolates, 2.5% (n = 5) were resistant to triclosan, multidrug resistance (MDR) and cross-resistance phenotypes were observed in these resistant strains, but not in susceptible strains. We did not observe any sense mutations within fabI gene in triclosan resistant strains. Moreover, except DC8603, all the others enhanced efflux pumps activity. Compared with ATCC 25922, except fabI, increased expression were also found in efflux pump encoding genes ydcV, ydcU, ydcS, ydcT, cysP, yihV, acrB, acrD and mdfA in studied strains with different PFGE patterns and STs types. Surprised, 5 susceptible E. coli isolates increased rapidly triclosan resistance only 4 days after exposure to subinhibitory triclosan concentration in vitro. Conclusions Our study is the first to be reported that short-term triclosan exposure in vitro increases triclosan resistance in susceptible E. coli isolates. Once strains have acquired resistance, they usually present MDR or cross-resistance phenotypes. Besides, our findings indicate that triclosan resistance were mainly involved by fabI overexpression in E. coli, and there was a close association between overexpression of efflux pumps with triclosan resistance.

scholarly journals Antibiotic Susceptibility Pattern of Escherichia coli Isolated from Various Clinical Samples in Duhok City, Kurdistan Region of Iraq

2020 ◽  
Vol 7 (3) ◽  
Author(s):  
Ibrahim A Naqid ◽  
Amer A Balatay ◽  
Nawfal Rasheed Hussein ◽  
Kurdistan Abdullah Saeed ◽  
Hiba Abdulaziz Ahmed ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
High Resistance ◽  
Antibiotic Susceptibility ◽  
Bacterial Infections ◽  
Antibiotic Sensitivity ◽  
Multidrug Resistant ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
Sensitivity Testing ◽  
E Coli

Background: Escherichia coli (E. coli) is one of the most common causative agents of bacterial infections. The emergence of multidrug-resistant E. coli is a major public health threat worldwide. Objectives: This study aimed to determine the antibiotic susceptibility profile of clinical isolates of E. coli from different samples. Methods: A total number of 454 clinical samples, including urine, wound, cervical swab, blood, semen, ascetic, and cerebral spinal fluid samples were collected from patients between January 2017 and February 2020. Then, E. coli was confirmed and susceptibility to different antibiotics was determined using the Vitek-2 compact system. Results: Escherichia coli isolates were more frequent in females (70.7%) than in males (29.3%). In the case of urine samples, E. coli was found to be highly susceptible to ertapenem (97.6%) and imipenem (96.4%) but resistant to ampicillin (87.8%). For wound and cervical swabs, E. coli was 100% resistant to ampicillin and cefepime but 100% sensitive to ertapenem and imipenem. It was found that E. coli isolates from blood samples were 100% resistant to ampicillin, ceftriaxone, and cefoxitin, and around 75% of them were sensitive to ertapenem, ciprofloxacin, and levofloxacin. Finally, E. coli isolated from other clinical samples were highly sensitive to ertapenem, imipenem, levofloxacin, nitrofurantoin, and cefazolin. Conclusions: Escherichia coli isolated from various clinical specimens showed differences in antibiotic sensitivity patterns, with high resistance to commonly used antibiotics. The most effective antibiotics against E. coli isolates were ertapenem, imipenem, and nitrofurantoin. However, the clinical isolates of E. coli displayed high resistance rates to ampicillin, ceftriaxone, and cefepime. Therefore, it is proposed to perform antibiotic sensitivity testing by physicians to select the most effective antibiotics.

Molecular Characterization of Antimicrobial Drug Resistance in Escherichia coli Isolated from Clinical Samples

2017 ◽  
Vol 8 (01) ◽  
Author(s):  
Ibtisam Habeeb AL-Azawi ◽  
Aqeel Reheeum Hassan ◽  
Alaa Hamza Jaber
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Clinical Samples ◽  
Chain Reaction ◽  
Antimicrobial Drug Resistance ◽  
Biochemical Characteristics ◽  
E Coli ◽  
Medical City ◽  
Polymerase Chain

A total of 49 different clinical samples (urine n=30, stool n=10, and blood n=8) were collected from patient admitted to the Al-Sadder medical City in Al-Najaf Governorate-Iraq. The results demonstrated that 49 specimens (100%) were diagnosed as E. coli by cultural, biochemical characteristics and Vitek2® system. Polymerase Chain Reaction has been used to detect of some genes which coding antimicrobial resistance in E. coli isolates. Regarding genes that responsible for ESBL enzymes (blaCTX-M, blaOXA and blaTEM), the current results proved that blaTEM genes have highest rate (97.95%) followed by blaTEM and blaOXA (93.75%) for each.

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