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Data Note

Data on antibiograms and resistance genes of Enterobacterales isolated from ready-to-eat street food of Ambato, Ecuador

[version 1; peer review: 3 approved]
PUBLISHED 17 Jun 2022
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This article is included in the Antimicrobial Resistance collection.

Abstract

Foodborne pathogens represent a significant cause of negative impacts on human health and the economy worldwide. Unfortunately, information about epidemiological insights in Latin American countries is scarce. The consumption of ready-to-eat street food in Ecuador is extensive, and information about the presence of foodborne pathogens, their virulence factors, and antimicrobial resistance is negligible. This data includes the occurrence, phenotypic antibiotic resistance profiles, and antibiotic resistance genes of Enterobacterales isolated from ready-to-eat street food in Ambato, central Ecuador during 2020 and 2021. The most common genera detected were Escherichia coli, Klebsiella spp., and Cronobacter spp. Agar disk diffusion assays were performed to determine their phenotypic resistance. The presence of antibiotic resistance genes conferring resistance against colistin, β-Lactams, aminoglycosides, tetracyclines, sulfonamides, fluoroquinolones, and amphenicols was detected via polymerase chain reaction (PCR) amplification.

Keywords

antibiotic resistance, enterobacterales, escherichia coli, street food, food microbiology

Introduction

This data contributes information about the antibiotic resistance profiles of Enterobacterales strains isolated from street food that will facilitate pathogen surveillance in Ecuador and Latin America. This data is useful for the scientific community to determine the presence of pathogenic Escherichia coli isolates and antibiotic resistance genes, including mobile colistin resistance genes, carbapenemases, quinolone resistance genes, and extended-spectrum β-lactamases present on Enterobacterales strains isolated from street food. Researchers and policymakers involved with the work related to the One Health initiative could also benefit from this data for retrospective and comparative analysis or epidemiological surveillance projects.1,2

Methods

Enterobacterales strains

Ready-to-eat street food was obtained in the streets of the city of Ambato, Ecuador, and processed the same day. A sharp sterile blade was used to cut the samples on sterile surfaces. 10 g of each sample was placed in sterile brain heart infusion broth (BHIB) (Merck, Darmstadt, Germany) in 90 ml, shaken on a rotator for 8-10 min, and incubated for 24 h at 37 °C. A large amount of broth was inoculated on MacConkey agar plates (Merck, Darmstadt, Germany), Cromocult Coliforms Agar (Merck KGaA, Darmstadt, Germany), and CHROMagar mSuperCARBA were incubated overnight at 37 °C under aerobic conditions. Further purification was performed on Macconkey agar.

The isolates were amplified by polymerase chain reaction (PCR), analysed using agarose gel electrophoresis and visualised with Sybr Safe DNA Gel Stain.3 For the identification of the isolated Enterobacterales, biochemical tests such as catalase, oxidase, TSI agar, Simmons citrate, lactose test, indole production, urea agar, methyl red test, and Voges-Proskauer were carried out and their interpretation was performed based on Bergey's manual.4 Additionally, the software for Automated Biometric Identification Systems (ABIS) was used to confirm the biochemical identification results.

Phenotypic antibiotic resistance profiles

Agar disk diffusion assays (Thermo Scientific Oxoid and Bioanalyse) on Mueller-Hinton Agar (Thermo Scientific Oxoid) were performed. Antibiograms tests were based on the measured diameter of the zones of inhibition and interpreted as sensible, intermediate or resistant by referring to CLSI breakpoints.5

Detection of E. coli pathotypes and antibiotic resistance genes detection via PCR

The PCR test was performed according to the standardized protocol of the UTA RAM One Health research group6,7: 2.5 μL of DNA from each sample and 22.5 μL of PCR mix containing 12.5 μL DreamTaq PCR Master Mix (ThermoFisher Scientific, USA), 9 μL Nuclease-free water, 0.5 μL Primer 1 and 0.5 μL Primer 2 (final concentration of primers: 0.5 μM) were mixed to run PCR. The PCR conditions are reported in Supplementary Table S4. PCR products were analyzed by 1.2% agarose gel electrophoresis stained by Sybr Safe DNA Gel Stain (ThermoFisher Scientific, USA).

Hierarchical clustering

Hierarchical clustering was performed using the Euclidean correlation method and clustered by affinity.2 The MeV Multiexperiment Viewer software version 4.8.1 was used in this study.

Dataset validation

The data presented show the frequency of isolation of Enterobacterales in 151 samples of ready-to-eat street food in Ambato, Ecuador (Figure 1). The specific characteristics (date of sampling, type of street food, location) of the samples were reported in Supplementary Table S1. A total of 145 isolates were analyzed, and the results of the biochemical tests were reported in Supplementary Table S2. Among them, 86 isolates corresponded to E. coli and 59 isolates to other Enterobacterales.

d5fe5175-b527-4e6d-af3f-be833b8f686e_figure1.gif

Figure 1. Occurrence of Enterobacterales on 151 samples of ready-to-eat street food in Ambato, Ecuador.

To visualize the relative similarity of the antimicrobial resistance patterns of the isolates, a hierarchical cluster analysis was performed using the results of the antibiograms, where the phenotypes ‘resistant’, ‘intermediate’, and ‘susceptible’ were observed as red, white, and blue colors respectively. Dendrograms and clustered data were assembled using the complete linkage method through Pearson correlation and sample leaf organization.7 For this purpose, the MeV Multiexperiment Viewer software version 4.8.1 was used.8 Figures 2 and 3 represent the resistance profiles and the hierarchical clustering of E. coli and the rest of Enterobacterales, respectively. The complete information is shown in Supplementary Table S3.

d5fe5175-b527-4e6d-af3f-be833b8f686e_figure2.gif

Figure 2. Profiles of antibiotic resistance and hierarchical tree of E. coli isolates.

Red: resistant, White: intermediate, Blue: sensitive.

d5fe5175-b527-4e6d-af3f-be833b8f686e_figure3.gif

Figure 3. Profiles of antibiotic resistance and a hierarchical tree of other Enterobacterales isolates.

Red: resistant, White: intermediate, Blue: sensitive.

Abbreviations: TE: Tetracycline 30 μg, AM: Ampicilin 10 μg, KF: cephalotin 30 μg, C: chloramphenicol 30 μg, CIP: Ciprofloxacin 5 μg, CTX: Cefatoxime 30 μg, LEV: Levofloxacin 5 μg, FOX: Cefoxitin 30 μg, STX: Trimethoprim/sulphamethoxazole 25 μg, AMC: Amoxicyllin/ClavulanicAcid 30 μg, CN: Gentamicin 10 μg, CRO: Ceftriaxione 30 μg, FEP: Cefepime 30 μg, ATM: Aztreonam 30 μg, IPM: Imipenem 10 μg, TPZ: Piperacillin/Tazobactam 110 μg, ETP: Ertapenem 10 μg, MEM: Meropenem 10 μg, CAZ: Ceftazidime 30 μg.

The presence of diarrheagenic E. coli pathotypes present in ready-to-eat food was assessed in this study through the analysis of virulence genes related to the pathotypes. Only one isolate (C2.1c) was positive for the eae gene, suggesting the potential presence of enteropathogenic E. coli (EPEC) or enterohemorrhagic E. coli (EHEC). The β-lactamase resistance genes of Enterobacterales isolated in this study are reported in Table 1. Mobile colistin resistance genes or quinolone resistance genes were not found in the Enterobacterales isolates. The complete information about virulence genes and antibiotic resistance genes are available in Supplementary Table S5. The information about primers and PCR conditions were shown in Supplementary Table S4. The gel electrophoresis images are available at Supplementary figure S6. The disk difussion assays figures were shown at Supplementary figure S7.

Table 1. Beta-lactamase resistance genes of Enterobacterales isolated from ready-to-eat food.

Ready-to-eat food samplesBacteriaSample IDDateLocation market streetsBeta-lactamases
blatemblacmy
Cow stomach stewE. coliM1.1a14/12/2020Mayorista+-
Cane juiceE. coliJc5.2c108/03/2021Modelo+-
Lupine cevicheE. colich5.1c208/03/2021Modelo+-
Cow stomach stewE. coliGap1a20/04/2021Sur-+
Boiled beans, ulluco and pork rindE. coliHmm1a10/05/2021Artesanal-+
Chilli sauceKlebsiella sppD1.2a07/12/2020Mayorista+-
Sweet meringue (espumilla)Salmonella sppE2.2b19/01/2021Primera de Mayo+-
Salad from street foodE. coliN1.2a20/12/2020Mayorista+-
Total Enterobacterales isolated6(145)2(145)

Data availability

Figshare project: https://figshare.com/projects/Data_on_antibiograms_and_resistance_genes_of_Enterobacterales_isolated_from_Ready-to-eat_street_food_of_Ambato_Ecuador/137014

This collection contains the following underlying data:

Figure 1. Occurrence of Enterobacterales on 151 samples of ready-to-eat street foods in Ambato, Ecuador. figshare. Figure. https://doi.org/10.6084/m9.figshare.19579087.v19

Table 1. Beta-lactamase resistance genes of Enterobacterales isolated from ready-to-eat food. figshare. Dataset. https://doi.org/10.6084/m9.figshare.19579099.v110

Figure 2 and 3. Antibiotic resistance profiles and hierarchical trees of Enterobacterales isolated from ready-to-eat street food in Ambato, Ecuador. figshare. Dataset. https://doi.org/10.6084/m9.figshare.19579267.v111

Extended data

This collection contains the following extended data:

Supplementary table S1. Characteristics (Sample type, date, treatment type, location, coordinates) of the ready-to-eat food samples. figshare. Dataset. https://doi.org/10.6084/m9.figshare.19579108.v112

Supplementary table S2. Biochemical tests performed on Enterobacterales isolates from Ready-to-eat Street Food in Ambato, Ecuador. figshare. Dataset. https://doi.org/10.6084/m9.figshare.19579177.v113

Supplementary table S3. Antibiogram of Enterobacterales isolated from ready-to-eat Street food of Ambato, Ecuador. figshare. Dataset. https://doi.org/10.6084/m9.figshare.19579189.v114

Supplementary table S4. Primers used in this study and PCR conditions. figshare. Dataset. https://doi.org/10.6084/m9.figshare.19579198.v115

Supplementary table S5. Antibiotic resistance genes and virulence genes harbored by Enterobacterales isolates from ready-to-eat street food in Ecuador. figshare. Dataset. https://doi.org/10.6084/m9.figshare.19579207.v116

Supplementary figure S6. PCR results (positive electrophoresis images). figshare. Figure. https://doi.org/10.6084/m9.figshare.19729618.v117

Supplementary figure S7. Disk diffusion assay images-Antibiotic resistance evaluation of Enterobacterales isolated from ready-to-eat street food of Ambato, Ecuador. figshare. Figure. https://doi.org/10.6084/m9.figshare.19729630.v118

Data are available under the terms of the Creative Commons Attribution 4.0 International license (CC-BY 4.0).

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Tubón J, Barragán-Fonseca G, Lalaleo L and Calero-Cáceres W. Data on antibiograms and resistance genes of Enterobacterales isolated from ready-to-eat street food of Ambato, Ecuador [version 1; peer review: 3 approved]. F1000Research 2022, 11:669 (https://doi.org/10.12688/f1000research.117116.1)
NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article.
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ApprovedThe paper is scientifically sound in its current form and only minor, if any, improvements are suggested
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Not approvedFundamental flaws in the paper seriously undermine the findings and conclusions
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PUBLISHED 17 Jun 2022
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Reviewer Report 19 Jul 2022
Erika A. Rodriguez, Bacterial Molecular Epidemiology Line, Research Group in Basic and Applied Microbiology (MICROBA), School of Microbiology, University of Antioquia, Medellín, Colombia 
Approved
VIEWS 7
The data note by Tubón J et al. aims to give information about the antibiotic resistance profiles of Enterobacterales strains isolated from street food in Ecuador. The work is relevant because, usually in Latin American countries, street food consumption is ... Continue reading
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HOW TO CITE THIS REPORT
Rodriguez EA. Reviewer Report For: Data on antibiograms and resistance genes of Enterobacterales isolated from ready-to-eat street food of Ambato, Ecuador [version 1; peer review: 3 approved]. F1000Research 2022, 11:669 (https://doi.org/10.5256/f1000research.128952.r141527)
NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article.
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Reviewer Report 05 Jul 2022
Edgar Gonzalez-Villalobos, Catalan Institute for Water Research (ICRA), Girona, Spain 
Approved
VIEWS 12
In Latin American countries the consumption of street food represents in the majority the alimentary base, due to the conditions that present the most countries belonging to Latin American, for this reason, the information generated by Turbón et al., could ... Continue reading
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CITE
HOW TO CITE THIS REPORT
Gonzalez-Villalobos E. Reviewer Report For: Data on antibiograms and resistance genes of Enterobacterales isolated from ready-to-eat street food of Ambato, Ecuador [version 1; peer review: 3 approved]. F1000Research 2022, 11:669 (https://doi.org/10.5256/f1000research.128952.r141528)
NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article.
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Reviewer Report 23 Jun 2022
Laura Sala-Comorera, UCD School of Biomolecular and Biomedical Science, UCD Earth Institute, Dublin, Ireland;  UCD Conway Institute, University College Dublin, Dublin, Ireland 
Approved
VIEWS 12
In this data note article, Turbón and colleagues examined the antibiograms and resistance genes profiles of Enterobacterales order strains isolated in ready-to-eat street food in Ecuador. A larger number of samples and types of food have been selected, resulting in ... Continue reading
CITE
CITE
HOW TO CITE THIS REPORT
Sala-Comorera L. Reviewer Report For: Data on antibiograms and resistance genes of Enterobacterales isolated from ready-to-eat street food of Ambato, Ecuador [version 1; peer review: 3 approved]. F1000Research 2022, 11:669 (https://doi.org/10.5256/f1000research.128952.r141529)
NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article.

Comments on this article Comments (0)

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Alongside their report, reviewers assign a status to the article:
Approved - the paper is scientifically sound in its current form and only minor, if any, improvements are suggested
Approved with reservations - A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit.
Not approved - fundamental flaws in the paper seriously undermine the findings and conclusions
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