Molecular detection of blaCTX-M gene to predict phenotypic cephalosporin resistance and clinical outcome of Escherichia coli bloodstream infections in Vietnam

Abstract Background Blood stream infections (BSI) caused by Extended Spectrum Beta-Lactamases (ESBLs) producing Enterobacteriaceae is a clinical challenge leading to high mortality, especially in developing countries. In this study, we sought to describe the epidemiology of ESBL-producing Escherichia coli strains isolated from Vietnamese individuals with BSI, to investigate the concordance of genotypic-phenotypic resistance, and clinical outcome of ESBL E. coli BSI. Methods A total of 459 hospitalized patients with BSI were screened between October 2014 and May 2016. 115 E. coli strains from 115 BSI patients were isolated and tested for antibiotic resistance using the VITEK®2 system. The ESBL phenotype was determined by double disk diffusion method following the guideline of Clinical and Laboratory Standards Institute. Screening for beta-lactamase (ESBL and carbapenemase) genes was performed using a multiplex-PCR assay. Results 58% (67/115) of the E. coli strains were ESBL-producers and all were susceptible to both imipenem and meropenem. Resistance to third-generation cephalosporin was common, 70% (81/115) were cefotaxime-resistant and 45% (52/115) were ceftazidime-resistant. blaCTX-M was the most common ESBL gene detected (70%; 80/115) The sensitivity and specificity of blaCTX-M-detection to predict the ESBL phenotype was 87% (76–93% 95% CI) and 54% (39–48% 95% CI), respectively. 28%% (22/80) of blaCTX-M were classified as non-ESBL producers by phenotypic testing for ESBL production. The detection of blaCTX-M in ESBL-negative E. coli BSI was associated with fatal clinical outcome (27%; 6/22 versus 8%; 2/26, p = 0.07). Conclusion A high prevalence of ESBL-producing E. coli isolates harbouring blaCTX-M was observed in BSI patients in Vietnam. The genotypic detection of blaCTX-M may have added benefit in optimizing and guiding empirical antibiotic therapy of E. coli BSI to improve clinical outcome.

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Antibiotic Resistance of Escherichia Coli Isolated from Broiler Chickens

Folia Veterinaria ◽

10.2478/fv-2019-0021 ◽

2019 ◽

Vol 63 (3) ◽

pp. 1-8

Author(s):

M. Sciberras ◽

M. Pipová ◽

I. Regecová ◽

P. Jevinová ◽

S. Demjanová

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Broiler Chickens ◽

Tetracycline Resistance ◽

Diffusion Method ◽

Disk Diffusion Method ◽

Phenotypic Resistance ◽

E Coli ◽

Mueller Hinton Agar ◽

Almost All

Abstract The purpose of this study was to detect the antibiotic resistance of forty-one Escherichia coli isolates from the intestinal contents of slaughtered broiler chickens using the disk diffusion method according to Kirby-Bauer. Mueller-Hinton agar plates were inoculated with 0.1 ml overnight broth cultures of individual E. coli isolates and the disks with the following concentrations of antibiotics were applied onto them: ampicillin (10 μg), cefotaxime (30 μg), gentamicin (10 μg), streptomycin (10 μg), azithromycin (15 μg), tetracycline (30 μg), ciprofloxacin (30 μg) and levofloxacin (3 μg). After the incubation at 37 °C for 16—18 hours, the inhibition zones were measured and interpreted in accordance with the Clinical and Laboratory Standard Institute (CLSI) zone diameter breakpoints. Almost all E. coli isolates showed resistance to tetracycline (92.68 %), most of them were resistant to gentamicin (75.61 %) and levofloxacine (70.73 %). Phenotypic resistance to tetracycline was further confirmed with the help of the Polymerase Chain Reaction (PCR) procedure focused on the presence of specific tet(A) and tet(B) genes. These genes were detected in all 41 E. coli isolates. On the contrary, E. coli isolates were highly susceptible to both azithromycin and streptomycin. In conclusion, the study highlighted the role of commensal E. coli bacteria isolated from the intestines of broiler chickens as an important reservoir of tetracycline resistance genes.

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Occurrence of Escherichia coli producing extended spectrum β-lactamases in food-producing animals

Acta Veterinaria Hungarica ◽

10.1556/004.2021.00036 ◽

2021 ◽

Author(s):

Bence Balázs ◽

József Bálint Nagy ◽

Zoltán Tóth ◽

Fruzsina Nagy ◽

Sándor Károlyi ◽

...

Keyword(s):

Escherichia Coli ◽

Gene Families ◽

Diffusion Method ◽

Disk Diffusion Method ◽

Chain Reactions ◽

E Coli ◽

Esbl Gene ◽

Source Of Infection ◽

Extended Spectrum ◽

Esbl Producers

Abstract Multidrug resistance due to the production of extended-spectrum beta-lactamases (ESBLs) is a major problem in human as well as in veterinary medicine. These strains appear in animal and human microbiomes and can be the source of infection both in animal and in human healthcare, in accordance with the One Health theorem. In this study we examined the prevalence of ESBL-producing bacteria in food-producing animals. We collected 100 porcine and 114 poultry samples to examine the prevalence of ESBL producers. Isolates were identified using the MALDI-TOF system and their antibiotic susceptibility was tested using the disk diffusion method. ESBL gene families and phylogroups were detected by polymerase chain reactions. The prevalence of ESBL producers was relatively high in both sample groups: 72 (72.0%) porcine and 39 (34.2%) poultry isolates were ESBL producers. Escherichia coli isolates were chosen for further investigations. The most common ESBL gene was CTX-M-1 (79.3%). Most of the isolates belong to the commensal E. coli phylogroups. The porcine isolates could be divided into three phylogroups, while the distribution of the poultry isolates was more varied. In summary, ESBL-producing bacteria are prevalent in the faecal samples of the examined food-producing animals, with a dominance of the CTX-M-1 group enzymes and commensal E. coli phylogroups.

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Detection of CTX-M-type ESBLs Escherichia coli at Universiti Kebangsaan Malaysia Medical Centre

Bangladesh Journal of Medical Science ◽

10.3329/bjms.v15i2.27849 ◽

2016 ◽

Vol 15 (2) ◽

pp. 257-261

Author(s):

Siti Norlia Othman ◽

Salasawati Hussin ◽

Ramliza Ramli ◽

MM Rahman

Keyword(s):

Escherichia Coli ◽

Medical Science ◽

Medical Centre ◽

Disk Diffusion ◽

Diffusion Method ◽

Phenotypic Characteristics ◽

Disk Diffusion Method ◽

Disk Diffusion Test ◽

E Coli ◽

Esbl Producers

Objective: To determine ESBLs and CTX-M-type ESBL-producing Escherichia coli at Universiti Kebangsaan Malaysia Medical Centre (UKMMC) by multiplex PCR.Materials and Methods: ESBL producing E. coli strains were confirmed by disk diffusion method. On the contrary, CTX-M-type ESBL-producing E. coli strains were confirmed by multiplex PCR.Results: Out of 554 collected E. coli isolates from UKMMC, 96 of these were detected as ESBL-producers. In 96 isolates, 76 viable strains were subjected to multiplex PCR for the detection of blaCTX genes. In which 70 (92.1%) were CTX-M-type ESBLs with the majority of CTX-M-1 group (77.1%), followed by CTX-M-9 group (21.4%) and one (1.4%) from CTX-M-2 group. Conclusions: CTX-M-type ESBLs were the predominant ESBL types isolated at UKMMC. CTX-M-1 and -9 groups were found in majority of the clinical isolates. Although phenotypic characteristics based on disk diffusion test provided similar results, however, molecular detection of genes of organisms is important for further epidemiological investigations.Bangladesh Journal of Medical Science Vol.15(2) 2016 p.257-261

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Pathotyping and Antibiotic Resistance Profiling of Escherichia coli Isolates from Children with Acute Diarrhea in Amatole District Municipality of Eastern Cape, South Africa

BioMed Research International ◽

10.1155/2020/4250165 ◽

2020 ◽

Vol 2020 ◽

pp. 1-10

Author(s):

S. A. Omolajaiye ◽

K. O. Afolabi ◽

B. C. Iweriebor

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Antimicrobial Agents ◽

Childhood Mortality ◽

Diffusion Method ◽

Resistance Pattern ◽

Disk Diffusion Method ◽

Phenotypic Resistance ◽

E Coli ◽

Diarrheagenic Escherichia Coli

Background. Diarrhea has been reported as the leading cause of childhood mortality and morbidity globally but with disproportionate impacts in developing nations. Among bacterial etiologic agents of diarrhea, diarrheagenic Escherichia coli is the main cause of the disease among children under the age of 5 years. This study is aimed at determining the prevalence and antibiogram pattern of diarrheagenic Escherichia coli (DEC) pathotypes associated with diarrhea cases in the study area. Methods. A total of 120 presumptive isolates of E. coli were obtained from diarrheal stool samples from male and female patients below 12 years of age using chromogenic agar. Confirmation of the isolates and screening for virulence genes were determined by polymerase chain reaction (PCR) while antimicrobial susceptibility testing was performed using the disk diffusion method. The presence of antibiotic resistance genes to chloramphenicol and tetracycline among the confirmed isolates was also profiled by PCR based on the observed phenotypic resistance pattern. Results. Of the 120 presumptive isolates, 88.3% (106/120) were confirmed as E. coli through PCR. The molecular pathotyping of the confirmed isolates showed their distribution as 41% (43/106) of diffusely adhering E. coli (DAEC), 17% (18/106) of enterohemorrhagic E. coli (EHEC), 17% (18/106) of enteropathogenic E. coli (EPEC), and 10% (11/106) of enteroinvasive E. coli (EIEC), while enteroaggregative E. coli (EAEC) and enterotoxigenic E. coli (ETEC) were not detected, and the remaining 15% did not belong to any pathotype. Notably, high resistance of the isolates to commonly used antimicrobials was observed as follows: ampicillin (98%), chloramphenicol (94%), trimethoprim-sulfamethoxazole (96%), and tetracycline (90.6%), while a relatively low number of the confirmed isolates were resistant to ciprofloxacin (45%) and imipenem (36%). In addition, 94% of the isolates that exhibited phenotypic resistance against chloramphenicol harbored the catA1 resistance gene while 89% that showed resistance to tetracycline had tetA genes. Conclusions. These findings showed that DEC could be considered as the leading etiologic bacterial agent responsible for diarrhea in the study community, and the observable high degree of resistance of the isolates to antimicrobial agents is of huge significance, calling for stakeholders to adopt and consolidate the existing antimicrobial stewardship scheme of the government, in order to ensure an uncompromised public health.

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Prevalence of Antibiotic-Resistant Escherichia coli Isolates from Local and Imported Retail Chicken Carcasses

Journal of Food Protection ◽

10.4315/jfp-20-113 ◽

2020 ◽

Vol 83 (12) ◽

pp. 2200-2208

Author(s):

NAHLA O. ELTAI ◽

HADI M. YASSINE ◽

TAHRA EL-OBEID ◽

SARA H. AL-HADIDI ◽

ASMAA A. AL THANI ◽

...

Keyword(s):

Escherichia Coli ◽

Multidrug Resistant ◽

Chicken Meat ◽

Diffusion Method ◽

Bacterial Strains ◽

Disk Diffusion Method ◽

Antibiotic Resistant ◽

E Coli ◽

Multiplex Pcr Assay ◽

Synergy Test

ABSTRACT The spread of antibiotic resistance among bacterial strains has been associated with consumption of food contaminated with both pathogenic and nonpathogenic bacteria. The objective of this study was to determine the prevalence of antibiotic resistant Escherichia coli isolates in local and imported retail raw chicken meat in Qatar. A total of 270 locally produced (chilled) and imported (chilled or frozen) whole chicken carcasses were obtained from three Hypermarket stores in Qatar. The 216 E. coli isolates recovered from the chicken samples were subjected to antibiotic susceptibility testing with the disk diffusion method. Extended-spectrum β-lactamase (ESBL) production was evaluated with the double disk synergy test. Isolates harboring colistin resistance were identified with a multiplex PCR assay and DNA sequencing. Nearly 89% (192) of the 216 isolates were resistant to at least one of the 18 antibiotics tested. Isolates from local and imported chicken carcasses had relatively higher resistance to sulfamethoxazole (62% of isolates), tetracycline (59.7%), ampicillin and trimethoprim (52.3% each), ciprofloxacin (47.7%), cephalothin (45.4%), and colistin (31.9%). Less resistance was found to amoxicillin–clavulanic acid (6%), ceftriaxone (5.1%), nitrofurantoin (4.2%), piperacillin-tazobactam (4.2%), cefepime (2.3%), meropenem (1.4%), ertapenem (0.9%), and amikacin (0.9%). Nine isolates (4.2%) were ESBL producers, and 137 (63.4%) were multidrug resistant. The percentages of multidrug-resistant, ESBL-producing, and colistin resistant isolates were significantly higher among isolates from local chilled than from imported chilled and frozen chicken samples. Our findings indicate the high prevalence of antibiotic-resistant E. coli in chicken meat sold at retail in Qatar. HIGHLIGHTS

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Phenotypic Antimicrobial Susceptibility of Escherichia coli from Raw Meats, Ready-to-Eat Meats, and Their Related Samples in One Health Context

Microorganisms ◽

10.3390/microorganisms9020326 ◽

2021 ◽

Vol 9 (2) ◽

pp. 326

Author(s):

Frederick Adzitey ◽

Nurul Huda ◽

Amir Husni Mohd Shariff

Keyword(s):

Escherichia Coli ◽

Antimicrobial Resistance ◽

Antimicrobial Susceptibility ◽

Resistance Index ◽

Diffusion Method ◽

Resistant Bacteria ◽

Disk Diffusion Method ◽

E Coli ◽

The Usa ◽

Raw Meats

Meat is an important food source that can provide a significant amount of protein for human development. The occurrence of bacteria that are resistant to antimicrobials in meat poses a public health risk. This study evaluated the occurrence and antimicrobial resistance of E. coli (Escherichia coli) isolated from raw meats, ready-to-eat (RTE) meats and their related samples in Ghana. E. coli was isolated using the USA-FDA Bacteriological Analytical Manual and phenotypic antimicrobial susceptibility test was performed by the disk diffusion method. Of the 200 examined meats and their related samples, 38% were positive for E. coli. Notably, E. coli was highest in raw beef (80%) and lowest in RTE pork (0%). The 45 E. coli isolates were resistant ≥ 50% to amoxicillin, trimethoprim and tetracycline. They were susceptible to azithromycin (87.1%), chloramphenicol (81.3%), imipenem (74.8%), gentamicin (72.0%) and ciprofloxacin (69.5%). A relatively high intermediate resistance of 33.0% was observed for ceftriaxone. E. coli from raw meats, RTE meats, hands of meat sellers and working tools showed some differences and similarities in their phenotypic antimicrobial resistance patterns. Half (51.1%) of the E. coli isolates exhibited multidrug resistance. The E. coli isolates showed twenty-two different resistant patterns, with a multiple antibiotic resistance index of 0.0 to 0.7. The resistant pattern amoxicillin (A, n = 6 isolates) and amoxicillin-trimethoprim (A-TM, n = 6 isolates) were the most common. This study documents that raw meats, RTE meats and their related samples in Ghana are potential sources of antimicrobial-resistant E. coli and pose a risk for the transfer of resistant bacteria to the food chain, environment and humans.

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Evaluation of extended spectrum beta lactamase enzymes prevalence in clinical isolates of Escherichia coli

Microbiology Research ◽

10.4081/mr.2011.e8 ◽

2011 ◽

Vol 2 (1) ◽

pp. 8

Author(s):

Ronak Bakhtiari ◽

Jalil Fallah Mehrabadi ◽

Hedroosha Molla Agamirzaei ◽

Ailar Sabbaghi ◽

Mohammad Mehdi Soltan Dallal

Keyword(s):

Escherichia Coli ◽

Disk Diffusion ◽

Confirmatory Test ◽

Diffusion Method ◽

Multi Drug Resistance ◽

Beta Lactamase ◽

Disk Diffusion Method ◽

Extended Spectrum Beta Lactamase ◽

E Coli ◽

Extended Spectrum

Resistance to b-lactam antibiotics by gramnegative bacteria, especially Escherichia coli (E. coli), is a major public health issue worldwide. The predominant resistance mechanism in gram negative bacteria particularly E. coli is via the production of extended spectrum beta lactamase (ESBLs) enzymes. In recent years, the prevalence of b-lactamase producing organisms is increased and identification of these isolates by using disk diffusion method and no-one else is not satisfactory. So, this investigation focused on evaluating the prevalence of ESBL enzymes by disk diffusion method and confirmatory test (Combined Disk). Five hundred clinical samples were collected and 200 E. coli isolates were detected by standard biochemical tests. To performing initial screening of ESBLs was used from Disk diffusion method on E. coli isolates. A confirmation test (Combined Disk method) was performed on isolates of resistant to cephalosporin's indicators. Up to 70% isolates exhibited the Multi Drug Resistance phenotype. In Disk diffusion method, 128(64%) E. coli isolates which resistant to ceftazidime and cefotaxime while in Combined Disk, among 128 screened isolates, 115 (89.8%) isolates were detected as ESBLs producers. This survey indicate beta lactamase enzymes are playing a significant role in antibiotic resistance and correct detection of them in phenotypic test by using disk diffusion and combined Disk is essential for accurate recognition of ESBLs.

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Occurrence of toxigenic Escherichia coli in raw milk cheese in Brazil

Arquivo Brasileiro de Medicina Veterinária e Zootecnia ◽

10.1590/s0102-09352007000200035 ◽

2007 ◽

Vol 59 (2) ◽

pp. 508-512 ◽

Cited By ~ 27

Author(s):

B.R. Paneto ◽

R.P. Schocken-Iturrino ◽

C. Macedo ◽

E. Santo ◽

J.M. Marin

Keyword(s):

Escherichia Coli ◽

Polymerase Chain Reaction ◽

Antimicrobial Agents ◽

Raw Milk ◽

Diffusion Method ◽

Chain Reaction ◽

Disk Diffusion Method ◽

E Coli ◽

Polymerase Chain ◽

Raw Milk Cheese

The occurrence of toxigenic Escherichia coli in raw milk cheese was surveyed in Middle Western Brazil. Fifty samples of cheese from different supermarkets were analyzed for E.coli. The isolates were serotyped and screened for the presence of verotoxigenic E. coli (VTEC) and enterotoxigenic E. coli (ETEC) by Polymerase Chain Reaction (PCR). The susceptibility to thirteen antimicrobial agents was evaluated by the disk diffusion method. E.coli were recovered from 48 (96.0%) of the samples. The serogroups identified were O125 (6.0%), O111 (4.0%), O55 (2.0%) and O119 (2.0%). Three (6.0%) and 1(2.0%) of the E.coli isolates were VTEC and ETEC, respectively. Most frequent resistance was observed to the following antimicrobials: cephalothin (60.0%), nalidixic acid (40.0%), doxycyclin (33.0%), tetracycline (31.0%) and ampicillin (29.0%).

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Molecular types, virulence profiles and antimicrobial resistance of Escherichia coli causing bovine mastitis

Veterinary Record Open ◽

10.1136/vetreco-2019-000369 ◽

2019 ◽

Vol 6 (1) ◽

pp. e000369 ◽

Cited By ~ 5

Author(s):

Magdalena Nüesch-Inderbinen ◽

Nadine Käppeli ◽

Marina Morach ◽

Corinne Eicher ◽

Sabrina Corti ◽

...

Keyword(s):

Escherichia Coli ◽

Bovine Mastitis ◽

Tetracycline Resistance ◽

Diffusion Method ◽

Sample Population ◽

Disk Diffusion Method ◽

E Coli ◽

Phylogenetic Grouping ◽

Resistance Rates ◽

Susceptibility Profiles

BackgroundEscherichia coli is an important aetiological agent of bovine mastitis worldwide.MethodsIn this study, 82 E. coli from bovine mastitis milk samples from 49 farms were analysed for their genetic diversity using phylogenetic grouping and multilocus sequence typing. The isolates were examined by PCR for a selection of virulence factors (VFs). Antimicrobial susceptibility profiles were assessed using the disk diffusion method.ResultsThe most prevalent phylogroups were group B1 (41.5 per cent of the isolates) and group A (30.5 per cent). A variety of 35 different sequence types (STs) were identified, including ST1125 (11 per cent), ST58 (9.8 per cent), ST10 (8.5 per cent) and ST88 (7.3 per cent). Aggregate VF scores (the number of unique VFs detected for each isolate) ranged from 1 to 3 for 63.4 per cent of the isolates and were at least 4 for 12.2 per cent. For 24.4 per cent of the isolates, the score was 0. The three most frequent VFs were traT, fyuA and iutA. The majority (72 per cent) of the isolates harboured traT. The majority (68.3 per cent) of the isolates were fully susceptible to all antimicrobials tested, with 22 per cent resistant to ampicillin and 14.6 per cent to tetracycline. Resistance rates were low for gentamicin (3.7 per cent), amoxicillin/clavulanic acid (2.4 per cent) and ceftiofur (1.2 per cent), respectively.ConclusionAmong the study’s sample population, E. coli strains were genotypically diverse, even in cows from the same farm, although some STs occurred more frequently than others. Susceptibility to clinically relevant compounds remained high.

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A Preliminary Study: Antibiotic Resistance Patterns of Escherichia coli and Enterococcus Species from Wildlife Species Subjected to Supplementary Feeding on Various South African Farms

Animals ◽

10.3390/ani10030396 ◽

2020 ◽

Vol 10 (3) ◽

pp. 396 ◽

Cited By ~ 1

Author(s):

Michaela Sannettha van den Honert ◽

Pieter Andries Gouws ◽

Louwrens Christiaan Hoffman

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Supplementary Feeding ◽

Health Concern ◽

Diffusion Method ◽

Disk Diffusion Method ◽

E Coli ◽

Wildlife Species ◽

Human Contact ◽

Resistance Patterns

Studies have shown that antibiotic resistance among wild animals is becoming a public health concern, owing to increased contact and co-habitation with domestic animals that, in turn, results in increased human contact, indirectly and directly. This type of farming practice intensifies the likelihood of antibiotic resistant traits in microorganisms transferring between ecosystems which are linked via various transfer vectors, such as rivers and birds. This study aimed to determine whether the practice of wildlife supplementary feeding could have an influence on the antibiotic resistance of the bacteria harboured by the supplementary fed wildlife, and thus play a potential role in the dissemination of antibiotic resistance throughout nature. Escherichia coli and Enterococcus were isolated from the faeces of various wildlife species from seven different farms across South Africa. The Kirby-Bauer disk diffusion method was used according to the Clinical and Laboratory Standards Institute 2018 guidelines. The E. coli (F: 57%; N = 75% susceptible) and Enterococcus (F: 67%; N = 78% susceptible) isolates from the supplementary fed (F) wildlife were in general, found to be more frequently resistant to the selection of antibiotics than from those which were not supplementary fed (N), particularly towards tetracycline (E. coli F: 56%; N: 71%/Enterococcus F: 53%; N: 89% susceptible), ampicillin (F: 82%; N = 95% susceptible) and sulphafurazole (F: 68%; N = 98% susceptible). Interestingly, high resistance towards streptomycin was observed in the bacteria from both the supplementary fed (7% susceptible) and non-supplementary fed (6% susceptible) wildlife isolates. No resistance was found towards chloramphenicol and ceftazidime.

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