scholarly journals Characterization of antimicrobial susceptibility, extended-spectrum β-lactamase genes and phylogenetic groups of Shigatoxin producing Escherichia coli isolated from patients with diarrhea in Iran

Author(s):  
Erfaneh Jafari ◽  
Mana Oloomi ◽  
Saeid Bouzari

Abstract Background Shiga toxin‐producing Escherichia coli (STEC) are among common foodborne bacterial pathogens and healthy livestock are the main source of this bacterium. Severe diseases attribute to two types of cytotoxin Stx1 and Stx2, which are also called Shiga toxin (Stx). Infection of humans with STEC may result in Acute diarrhea with or without bleeding, hemorrhagic colitis (HC) and the hemolytic uremic syndrome (HUS). As antibiotic resistance is increasingly being reported among STEC isolates obtained from livestock and patients worldwide, in this study the pattern of antibiotic resistance in clinical isolates was determined. Methods Stool samples were collected from patients with diarrhea. All samples were cultured and identified by biochemical and molecular tests. Antimicrobial susceptibility test and assessment of extended-spectrum β-lactamase (ESBL)-related genes were conducted. Moreover, phylogenetic groups were analyzed using quadruplex PCR, and DNA analysis assessed multi-locus sequence types (MLST). Results Out of 340 E. coli samples, 174 were identified as STEC by PCR. Antimicrobial susceptibility test results showed that, 99.4%, 96% and 93.1% of isolates were susceptible to imipenem/ertapenem, piperacillin–tazobactam and amikacin, respectively. The highest resistance was towards ampicillin (68.4%), followed by trimethoprim–sulfamethoxazole (59.8%), and tetracycline (57.5%). A total of 106 (60.9%) isolates were multidrug resistance (MDR) and 40.8% of isolates were determined to be extended spectrum β-lactamase producers. In 94.4% of isolates, genes responsible for ESBL production could be detected, and blaTEM was the most prevalent, followed by blaCTX-M9. Furthermore, phylogenetic grouping revealed that majority of STEC strains belonged to Group C, followed by Groups E, B2 and A. MLST unveiled diverse ST types. Conclusion A periodical surveillance studies and thorough understanding of antibiotic resistant profiles in STEC isolates could help select effective antibiotic treatment for patients and develop strategies to effectively manage food contamination and human infections.

2020 ◽  
Vol 19 (6) ◽  
pp. 1285-1293
Author(s):  
Lorina I. Badger-Emeka ◽  
Zainab Yaseen Al-Jaziri ◽  
Naheed Kausar ◽  
Nora Ahmad Al-Muhainy ◽  
Edric Estrella

Purpose: To investigate the antimicrobial susceptibility and extended spectrum β-lactamase (ESBL) production by clinical isolates of Escherichia coli (ESBL-EC) and Klebsiella species (ESBL-KP) associated with blood stream infections (BSIs).Methods: Bacteria isolation and identification were carried out using basic bacteriological and biochemical techniques. Antimicrobial susceptibility test was performed according to the guidelines of the Clinical Laboratory Standard Institute (CLSI). Vitek 2 Compact automated system was used toconfirm the identifications (ID) and antimicrobial susceptibility test (AST). The ESBL produced by E. coli and K. pneumoniae isolates were  phenotypically characterised using Modified Double Disc Synergy Test (MDDST), as recommended by CLSI.Results: All (100 %) isolates were sensitive to imipenem and meropenem, while susceptibility to other antibiotics varied. ESBL genotypes, viz, blaTEM, blaSHV and blaCTX were encountered in the BSIs. For ESBL-EC, TEM, SHV and CTX producers accounted for 33, 16.67 and 58.3 %,  respectively, while 75, 91.7 and 100 % of ESBL-KP were TEM, SHV and CTX producers, respectively. In ESBL-KP, 67 % coexisted with all three genotypes (blaTEM, blaSHV, and blaCTX, while 8.3 % of ESBL-EC coexisted with the three encoding genes (CTX, SHV and TEM).Conclusion: ESBL E. coli and K. pneumoniae associated with BSI have been identified as TEM, SHV and CXT producers, with more ESBL-KP coexisting with all three than ESBL-EC. Keywords: β-Lactamases, Genes, Klebsiella pneumoniae, Escherichia coli


Author(s):  
Doğan Akdoğan ◽  
Mustafa Güzel ◽  
Orhan Akpınar ◽  
Altan Aksoy ◽  
Yasemin Genç Bahçe

Aims: The lack of information about the inter variability of the test results obtained by CLSI and EUCAST requires further clarifications to interpret antimicrobial susceptibility patterns better. This study aimed to compare the CLSI and EUCAST interpretations of the antimicrobial susceptibility test results of the ESBL–producing uropathogenic Escherichia coli and Klebsiella pneumonia strains. Methods: After obtaining 157 ESBL-producing E. coli and 95, ESBL-producing K. pneumonia isolates from the urine specimens of the patients, Kirby-Bauer’s disc diffusion method was used for conducting antimicrobial susceptibility test. The test procedures and the interpretation of the results were carried out according to the year 2017 versions of both of the two guidelines. For the statistical comparison of concordance between the two guidelines, the Kappa coefficients and the concordance rates were calculated. Results: The results were graded in the range from perfect to poor agreement. For E. coli, interpretations of the AST results revealed a moderate to perfect agreement between both methods. Weighted Kappa agreement scores in the range from 0.42 to 1. The agreement for AMC, TPZ30/6, ceftazidime 10, meropenem, and aztreonam was poor without any inconsistencies. For Klebsiella, the kappa agreement score was in the range from 0.25 to 1. It was incompatible with AMC, TPZ 30/6, ceftazidime 10, aztreonam; there was poor agreement for cefepime, amikacin and ertapenem. Conclusions: Our results showed agreement between the two guidelines for uropathogenic extended-spectrum ß-lactamase producing Escherichia coli and Klebsiella pneumoniae but also showed inconsistencies between two guidelines. Therefore, the results of our study contribute to the comparison of these guidelines for interpreting antibiotic susceptibilities.


2018 ◽  
Vol 38 (11) ◽  
pp. 2150-2154 ◽  
Author(s):  
Ruben V. Horn ◽  
Windleyanne G.A. Bezerra ◽  
Elisângela S. Lopes ◽  
Régis S.C. Teixeira ◽  
Isaac N.G. Silva ◽  
...  

ABSTRACT: This study aimed to isolate Escherichia coli and Salmonella enterica from captured feral pigeons in Fortaleza, Brazil, and, in addition to evaluate the antimicrobial susceptibility profiles and diagnose diarrheagenic E. coli strains. Pigeons were captured in four public locations in Fortaleza with three techniques. Individual cloacal swab samples were collected and submitted to bacterial isolation, biochemical identification and antimicrobial susceptibility test. Disk diffusion technique was used with twelve antibiotics. E. coli strains were submitted to DNA extraction followed by PCR to diagnose five diarrheagenic pathotypes. A total of 124 birds were captured. One bird was positive for Salmonella enterica (0.81%) and 121 (97.58%) were positive for E. coli. Among these, 110 isolates were submitted to antimicrobial susceptibility test and 28.18% (31/110) presented resistance to at least one antibiotic. Resistance to azithromycin was the most frequent (21.82%), followed by tetracycline (10.91%) and sulfamethoxazole with trimethoprim (8.9%). Multidrug resistance, calculated as a resistance to at least 3 antimicrobial classes, was identified in 3.64% (4/110) of strains. The maximum number of antimicrobial classes to which one strain was resistant was seven. Results demonstrated nine different resistance profiles and the most frequent was tetracycline and sulfamethoxazole with trimethoprim (4 strains), followed by chloramphenicol, azithromycin, tetracycline and sulfamethoxazole with trimethoprim (3 strains). Amoxicillin with clavulanic acid and tobramycin presented lowest levels of antimicrobial resistance, to which none of the tested strains were resistant. A single strain was positive for the eltB gene, which is a diagnostic tool to identify the Enterotoxigenic E. coli (ETEC) pathotype. None of the other investigated genes (stx1, stx2, estA, eaeA, ipaH, aatA and aaiC) were identified. The single isolate of S. enterica was a rough strain of Salmonella enterica subsp. enterica, but serotype identification was not possible. However, this isolate presented resistance to amoxicillin, amoxicillin with clavulanic acid, tetracycline and sulfamethoxazole with trimethoprim. Therefore, captured feral pigeons of Fortaleza presented a low prevalence of S. enterica and diarrheagenic E. coli. Considering the investigated pathogens, our results suggest a good health status and a low public health risk. However, important antimicrobial resistance profiles were identified.


2021 ◽  
Vol 33 ◽  
pp. 06009
Author(s):  
Anindya Dwi Ash-Santri ◽  
Vinsa Cantya Prakasita ◽  
Yosua Kristian Adi ◽  
Teguh Budipitojo ◽  
Agnesia Endang Tri Hastuti Wahyuni

Atelerix albiventris and Hystrix javanica are currently traded as pets or consumed in Indonesia, but there has been no research about bacteria from the vulva swab before. This research aims to isolate and identify bacteria from the vulva swabs of Atelerix albiventris and Hystrix javanica, and identify their antibiotic susceptibility. Samples were isolated by blood agar plates and selective media and identified by biochemical tests. Kirby Bauer’s disk diffusion method was used for the antimicrobial susceptibility test. The result showed that from Atelerix albiventris was isolated and identified Escherichia coli and Proteus mirabilis, while from Hystrix javanica was isolated and identified Escherichia coli. The identified Escherichia coli was sensitive to Amikacin, Amoxycillin, Ampicillin, Enrofloxacin, Fosfomycin, Chloramphenicol, Tetracycline, Trimethoprim, and Kanamycin; intermediate to Streptomycin; and resistant to Erythromycin and Penicillin G. The identified Proteus mirabilis was sensitive to Amikacin, Amoxycillin, Kanamycin, Enrofloxacin, and Fosfomycin; and resistant to Erythromycin, Penicillin G, Streptomycin, Ampicillin, Chloramphenicol, Tetracycline, and Trimethoprim. This research concludes that Escherichia coli and Proteus mirabilis were isolated from vulva swab of Atelerix albiventris and Hystrix javanica, Escherichia coli was sensitive to eight antibiotics, while Proteus mirabilis was sensitive to five antibiotics.


2021 ◽  
Vol 2021 ◽  
pp. 1-5
Author(s):  
Mojtaba Moosavian ◽  
Mahtab Khoshkholgh Sima ◽  
Nazanin Ahmad Khosravi ◽  
Effat Abbasi Montazeri

Antibiotic resistance mechanisms in Enterobacteriaceae are causative agents of global health problems. Bacterial infections due to multidrug resistance (MDR) may be mediated by the overexpression of efflux pumps. In this study, we investigated the prevalence of oqxA and oqxB genes as two encoding agents of efflux pumps and the determination of antibiotic resistance rate in clinical isolates of Enterobacteriaceae. In this study, 100 Enterobacteriaceae isolates collected from different clinical specimens of infectious patients, such as wounds, urine, blood, discharge, and abscesses except stool, were examined. Identification of the isolates was performed using standard biochemical tests such as TSI, citrate, urea, lysine, SIM, MR-VP, and gas production. The antimicrobial susceptibility test was carried out by the Kirby–Bauer disk diffusion method according to CLSI guidelines, and finally, the oqxA and oqxB genes were detected by the PCR method. Among 100 Enterobacteriaceae isolates, Escherichia coli and Enterobacter gergoviae were the most common isolates with 71% and 20%, respectively. Also, the lowest isolates belonged to Enterobacter cloacae (3%) and Klebsiella pneumoniae (1%). Out of 100 Enterobacteriaceae isolates, 37 isolates (37%) were positive for at least one of oqxA or oqxB genes, while both of these genes were detected among 12% of them. oqxAB genes were detected in 8 cases of 20 (40%) Enterobacter gergoviae and 4 cases of 71 (5.7%) E. coli isolates. The antimicrobial susceptibility test showed that all isolates (100%) were susceptible to imipenem, while the maximum resistance to piperacillin, ceftriaxone, and cefotaxime were 69%, 55%, and 55%, respectively. Also, the results of this study showed that antibiotic resistance in Enterobacteriaceae isolates caused by oqxAB genes is increasing among patients in Iran. Therefore, identification of resistant isolates and antibiotic monitoring programs are essential to prevent the spread of MDR isolates.


2019 ◽  
Vol 12 (8) ◽  
pp. 1195-1203 ◽  
Author(s):  
Djanette Barour ◽  
Amine Berghiche ◽  
Nadji Boulebda

Background and Aim: Lack of information about the antibiotic resistance in commensal Escherichia coli from Algerian livestock prompted us to do this study to determine the different levels of antimicrobial susceptibility, antibiotic multidrug resistance (MDR) rates, and phenotypical patterns of E. coli strains isolated from healthy cattle to control the spread of animal-resistant strains to humans and the environment. Materials and Methods: A total of 198 cattle were sampled (swabbed in the rectum), reared in the farms of Souk Ahras, Tebessa, and Oum el Bouaghi governorates of Eastern Algeria. Isolation of E. coli strains was performed on MacConkey agar and then the different strains were identified to the species level using an API 20E identification kit. Antimicrobial susceptibility was determined using a panel of 13 antibiotic disks by disk diffusion method on Mueller-Hinton agar. The double-disk synergy test with cefotaxime and amoxicillin-clavulanate disks was used for the screening of extended-spectrum beta-lactamase phenotypes. For colistin susceptibility, the minimum inhibitory concentration was examined using broth microdilutions technique. Results: The results showed that among the 198 E. coli isolates, elevated resistance rates were observed for ampicillin (59.09%) and tetracycline (43.43%), and moderate resistance rates for cephalothin (16.16%), trimethoprim/sulfamethoxazole (15.15%), and amoxicillin/clavulanate (11.62%); however, low resistance rates were found for nalidixic acid (8.08%), ciprofloxacin (7.07%), kanamycin (6.56%), cefotaxime (4.54%), chloramphenicol (4.04%), nitrofurantoin (2.52%), cefoxitin (2.02%), gentamycin (1.01%), and no resistance to colistin. However, nine extended-spectrum β-lactamases producing E. coli strains were identified. Forty-four different patterns were determined, indicating a wide variety of resistance, ranging from one antimicrobial to a combination of 10. Analysis of coresistances revealed that 63 isolates (31.82%) were susceptible to all antibiotics used in the study, 42 isolates (21.21%) were resistant to one antibiotic, 43 isolates (21.72%) were resistant to two antibiotics, 24 isolates (12.12%) resistant to three antibiotics, 26 isolates (13.13%) were resistant for more than three agents, and 45 isolates (22.73%) were MDR (which means resistant to three or more families of antibiotics). Conclusion: This study demonstrates that commensal E. coli remains a potential source of antibiotic resistance in view of the high prevalence of antimicrobial resistance. The vast range of MDR phenotypes, especially extended-spectrum β-lactamases producing strains, emphasizes the urgent requirement to adopt measures to control the use of antimicrobials, in particular, by private veterinarians, as well as the strengthening of veterinary surveillance networks for antimicrobial resistance to control the spread of MDR bacteria from animals to humans and the environment.


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