Antimicrobial Resistance Genes and Diversity of Clones among ESBL- and Acquired AmpC-Producing Escherichia coli Isolated from Fecal Samples of Healthy and Sick Cats in Portugal

The aim of the study was to analyze the mechanisms of resistance in extended-spectrum beta-lactamase (ESBL)- and acquired AmpC (qAmpC)-producing Escherichia coli isolates from healthy and sick cats in Portugal. A total of 141 rectal swabs recovered from 98 sick and 43 healthy cats were processed for cefotaxime-resistant (CTXR) E. coli recovery (in MacConkey agar supplemented with 2 µg/mL cefotaxime). The matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) method was used for E. coli identification and antimicrobial susceptibility was performed by a disk diffusion test. The presence of resistance/virulence genes was tested by PCR sequencing. The phylogenetic typing and multilocus sequence typing (MLST) were determined by specific PCR sequencing. CTXRE. coli isolates were detected in seven sick and six healthy cats (7.1% and 13.9%, respectively). Based on the synergy tests, 11 of 13 CTXRE. coli isolates (one/sample) were ESBL-producers (ESBL total rate: 7.8%) carrying the following ESBL genes: blaCTX-M-1 (n = 3), blaCTX-M-15 (n = 3), blaCTX-M-55 (n = 2), blaCTX-M-27 (n = 2) and blaCTX-M-9 (n = 1). Six different sequence types were identified among ESBL-producers (sequence type/associated ESBLs): ST847/CTX-M-9, CTX-M-27, CTX-M-1; ST10/CTX-M-15, CTX-M-27; ST6448/CTX-M-15, CTX-M-55; ST429/CTX-M-15; ST101/CTX-M-1 and ST40/CTX-M-1. Three of the CTXR isolates were CMY-2-producers (qAmpC rate: 2.1%); two of them were ESBL-positive and one ESBL-negative. These isolates were typed as ST429 and ST6448 and were obtained in healthy or sick cats. The phylogenetic groups A/B1/D/clade 1 were detected among ESBL- and qAmpC-producing isolates. Cats are carriers of qAmpC (CMY-2)- and ESBL-producing E. coli isolates (mostly of variants of CTX-M group 1) of diverse clonal lineages, which might represent a public health problem due to the proximity of cats with humans regarding a One Health perspective.

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Escherichia coli Producing Extended-Spectrum β-lactamases (ESBL) from Domestic Camels in the Canary Islands: A One Health Approach

Animals ◽

10.3390/ani10081295 ◽

2020 ◽

Vol 10 (8) ◽

pp. 1295

Author(s):

Isabel Carvalho ◽

María Teresa Tejedor-Junco ◽

Margarita González-Martín ◽

Juan Alberto Corbera ◽

Vanessa Silva ◽

...

Keyword(s):

Escherichia Coli ◽

Canary Islands ◽

Tetracycline Resistance ◽

Phylogenetic Groups ◽

Esbl Producer ◽

Carriage Rate ◽

Genetic Characteristics ◽

Disk Diffusion Test ◽

E Coli ◽

Specific Pcr

Objective: This work aimed to determine the carriage rate of ESBL-producing Escherichia coli as well as their genetic characteristics in camels from the Canary Islands, Spain. Methods: Fecal samples were recovered from 58 healthy camels from Gran Canaria (n = 32) and Fuerteventura Islands (n = 26) during July 2019. They were seeded on MacConkey (MC) agar no supplemented and supplemented (MC + CTX) with cefotaxime (2 µg/mL). Antimicrobial susceptibility was determined by disk diffusion test (CLSI, 2018). The presence of blaCTX-M, blaSHV, blaTEM,blaCMY-2 and blaOXA-1/48 genes was tested by PCR/sequencing. Furthermore, the mcr-1 (colistin resistance), tetA/tetB (tetracycline resistance), int1 (integrase of class 1 integrons) and stx1,2 genes were analyzed. Phylogenetic groups and sequence types were determined by specific-PCR/sequencing for selected isolates. Results: E. coli was obtained from all the 58 camels in MC media (100%) and in five of them in MC + CTX media (8.6%). Furthermore, 63.8% of E. coli isolates recovered from MC agar were susceptible to all the antibiotics tested. The five E. coli isolates recovered from MC + CTX media were characterized and two of them were ESBL-producers (3.4%). Both ESBL-producer isolates carried the blaCTX-M-15 gene and belonged to the lineages ST3018 (phylogroup A) and ST69 (phylogroup B1). The 3 ESBL-negative isolates recovered from MC-CTX plates were ascribed to phylogroup-B1. Conclusions: Camels can be a source of ESBL-producer bacteria, containing the widespread blaCTX-M-15 gene associated with the lineages ST3018 and ST69.

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Distribution of ciprofloxacin-resistance genes among ST131 and non-ST131 clones of Escherichia coli isolates with ESBL phenotypes isolated from women with urinary tract infection

Iranian Journal of Microbiology ◽

10.18502/ijm.v13i3.6389 ◽

2021 ◽

Author(s):

Masoumeh Rasoulinasab ◽

Fereshteh Shahcheraghi ◽

Mohammad Mehdi Feizabadi ◽

Bahram Nikmanesh ◽

Azade Hajihasani ◽

...

Keyword(s):

Escherichia Coli ◽

Resistance Rate ◽

Fluoroquinolone Resistance ◽

Double Mutation ◽

Extended Spectrum Beta Lactamase ◽

E Coli ◽

Specific Pcr ◽

Ciprofloxacin Resistance ◽

Chromosomal Mutations ◽

Esbl Producers

Background and Objectives: Escherichia coli (E. coli) sequence type 131 (ST131) is associated with extended-spectrum beta-lactamase (ESBL) production and fluoroquinolone resistance. This study aimed to investigate the prevalence of ST131, ESBL, and plasmid-mediated quinolone resistance (PMQR) genes in the ciprofloxacin-resistant (CIPR ) and ESBL producers from women with UTI. Materials and Methods: The CIP-resistant ESBL producing (CIPR /ESBL+ ) E. coli isolates were screened for ST131-by specific PCR of mdh and gyrB. The ESBL and PMQR genes were screened by single PCR. The ST131 and non-ST131 isolates were selected to determine the mutations of gyrA and parC using PCR and sequencing, and also their genetic background by the Pasteur-MLST scheme. Results: Overall, 55% (33/60) CIPR /ESBL+ isolates were identified as ST131 (94% O25b-ST131). Resistance rate to ampicillin-sulbactam (70%), aztreonam (97%) and gentamicin (61%), the prevalence of aac(6′)-Ib-cr (66%), blaCTX-M-15 (82%), the profile of qnrS+aac(6′)-Ib-cr (30%), and the double mutation in the parC was significantly higher in ST131 than nonST131 isolates. The coexistence of PMQR and ESBL genes was found in more than 50% of ST131 and non-ST131 isolates. ST131 isolates differentiated into PST43 and PST506. Conclusion: Management of women with UTI caused by the CIPR /ESBL+ isolates (ST131) co-harbored PMQR, ESBL, and chromosomal mutations, is important for their effective therapy.

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Multidrug-Resistant, Including Extended-Spectrum Beta Lactamase-Producing and Quinolone-Resistant, Escherichia coli Isolated from Poultry and Domestic Pigs in Dar es Salaam, Tanzania

Antibiotics ◽

10.3390/antibiotics10040406 ◽

2021 ◽

Vol 10 (4) ◽

pp. 406

Author(s):

Zuhura I. Kimera ◽

Fauster X. Mgaya ◽

Gerald Misinzo ◽

Stephen E. Mshana ◽

Nyambura Moremi ◽

...

Keyword(s):

Escherichia Coli ◽

Multidrug Resistant ◽

Antibiotics Resistance ◽

Beta Lactamase ◽

Domestic Pigs ◽

Extended Spectrum Beta Lactamase ◽

Phenotypic Profile ◽

E Coli ◽

Extended Spectrum ◽

Esbl Producers

We determined the phenotypic profile of multidrug-resistant (MDR) Escherichia coli isolated from 698 samples (390 and 308 from poultry and domestic pigs, respectively). In total, 562 Enterobacteria were isolated. About 80.5% of the isolates were E. coli. Occurrence of E. coli was significantly higher among domestic pigs (73.1%) than in poultry (60.5%) (p = 0.000). In both poultry and domestic pigs, E. coli isolates were highly resistant to tetracycline (63.5%), nalidixic acid (53.7%), ampicillin (52.3%), and trimethoprim/sulfamethoxazole (50.9%). About 51.6%, 65.3%, and 53.7% of E. coli were MDR, extended-spectrum beta lactamase-producing enterobacteriaceae (ESBL-PE), and quinolone-resistant, respectively. A total of 68% of the extended-spectrum beta lactamase (ESBL) producers were also resistant to quinolones. For all tested antibiotics, resistance was significantly higher in ESBL-producing and quinolone-resistant isolates than the non-ESBL producers and non-quinolone-resistant E. coli. Eight isolates were resistant to eight classes of antimicrobials. We compared phenotypic with genotypic results of 20 MDR E. coli isolates, ESBL producers, and quinolone-resistant strains and found 80% harbored blaCTX-M, 15% aac(6)-lb-cr, 10% qnrB, and 5% qepA. None harbored TEM, SHV, qnrA, qnrS, qnrC, or qnrD. The observed pattern and level of resistance render this portfolio of antibiotics ineffective for their intended use.

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Comparative analysis of genetic characterization of β-lactam-resistant Escherichia coli from bulk tank milk in Korea

Irish Veterinary Journal ◽

10.1186/s13620-021-00203-4 ◽

2021 ◽

Vol 74 (1) ◽

Author(s):

Hye-Ri Jung ◽

Koeun Kim ◽

Young Ju Lee

Keyword(s):

Escherichia Coli ◽

Clinical Mastitis ◽

Phylogenetic Groups ◽

Bulk Tank Milk ◽

Genetic Characteristics ◽

E Coli ◽

Intestinal Infections ◽

Antimicrobial Resistance Genes ◽

Bulk Tank ◽

Genotypic Characteristics

Abstract Background This study was conducted to analyze the genetic characteristics of 41 β-lactam-resistant Escherichia coli isolates, which are one of the common causes of environmental mastitis, isolated from the bulk tank milk of 290 dairy farms in five factories operated by three dairy companies in Korea. Results Analysis of the phenotypic and genotypic characteristics of β-lactam-resistant E. coli isolates revealed differences between factories even within the same company. Isolates from factory A1 and C1 showed high resistance to cephalothin (76.9 and 100%, respectively), which is a first-generation cephalosporins, whereas resistance to tetracycline was showed by only the isolates from factories B1 (60.0%), C2 (66.7%), and C3 (100%). Although all the 41 β-lactam-resistant E. coli isolates were positive for blaOXA-1, blaTEM-1 was highly prevalent in isolates from factories C2 (100%) and C3 (100%). Among 17 isolates resistant to both β-lactams and aminoglycosides, the most common multilocus sequence type was ST399 (13isolates, 76.5%). Furthermore, 2 (11.8%) and 12 (70.6%) isolates belonged to the phylogenetic groups B2 and D, respectively, which are invasive strains that cause intestinal infections, respectively. The predominant serogroup was O15 (70.6%), which is a globally distributed extraintestinal pathogen. Interestingly, one isolate from factory A1 belonged to O157 and carried six virulence genes, simultaneously. Conclusions Although E. coli isolates were isolated from bulk tank milk, and not the clinical mastitis samples, the presence of the phylogenetic groups B2 and D, and the serogroups O15 and O157, which harbor antimicrobial resistance genes and virulence factors, can pose a threat to public health.

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Prevalence of extended-spectrum beta-lactamase-producing Escherichia coli strains in dairy farm wastewater in Chiang Mai

Veterinary Integrative Sciences ◽

10.12982/vis.2021.030 ◽

2021 ◽

Vol 19 (3) ◽

pp. 349-362

Author(s):

Prayuth Saekhow ◽

◽

Chayaphon Sriphannam ◽

◽

Keyword(s):

Escherichia Coli ◽

Dairy Farm ◽

Pcr Amplification ◽

Beta Lactamase ◽

Chiang Mai ◽

Phylogenetic Groups ◽

Extended Spectrum Beta Lactamase ◽

E Coli ◽

Extended Spectrum ◽

Farm Wastewater

We investigated the prevalence of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli strains in dairy farm wastewater in Chiang Mai, Thailand. We analyzed wastewater samples collected from 150 dairy farms and found that 88.7% of the farms (n = 133) were positive for ESBL-producing E. coli. Multiplex polymerase chain reaction (PCR) amplification was performed to characterize the presence of bla CTX-M, bla TEM, and blaSHV in ESBL-producing isolates. blaCTX-M was found in all isolates (n = 133), followed by blaTEM (80/133, 60.2%), whereas blaSHV was not detected in any isolate. blaCTX-M and blaTEM were present in 60.2% (80/133) of the isolates, and 39.8% (53/133) isolates carried bla CTX-M alone. Subgroup analysis showed that CTX-M-1 was the most prevalent subgroup among the isolates (129/133, 97.0%), followed by CTX-M-8 (2/133, 1.5%) and CTX-M-9 (2/133, 1.5%). The distribution of the phylogenetic groups was as follows: group A (100/133, 75.2%), followed by B1 (14/133, 10.5%), D (6/133, 4.5%), F (6/133, 4.5%), B2 (4/133, 3.0%), and E (3/133, 2.3%). Based on enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) and dendrogram analysis, 24 isolates were classified into clades I (n = 21), II (n =1), and III (n =2). Minor genetic differences were found in all clade I isolates. Our data suggest that the circulating of ESBL-producing E. coli carried at least one bla gene strain distributed in dairy farm wastewater in Chiang Mai.

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Characterization of commensal Escherichia coli isolates from slaughtered sheep in Mexico

The Journal of Infection in Developing Countries ◽

10.3855/jidc.14001 ◽

2021 ◽

Vol 15 (11) ◽

pp. 1755-1760

Author(s):

Jorge Acosta-Dibarrat ◽

Edgar Enriquez-Gómez ◽

Martín Talavera-Rojas ◽

Edgardo Soriano-Vargas ◽

Armando Navarro ◽

...

Keyword(s):

Escherichia Coli ◽

Antimicrobial Resistance ◽

Multidrug Resistant ◽

Phylogenetic Group ◽

Phylogenetic Groups ◽

E Coli ◽

New Information ◽

Antimicrobial Resistance Genes ◽

Uremic Syndrome

Introduction: Commensal Escherichia coli is defined as bacteria without known virulence factors that could be playing a specific role in some diseases; however, they could be responsible to disseminate antimicrobial resistance genes to other microorganisms. This study aimed to characterize the commensal E. coli isolates obtained from slaughtered sheep in the central region of Mexico. Methodology: Isolates were classified as commensal E. coli when distinctive genes related to diarrheagenic pathotypes (stx1, stx2, eae, bfp, LT, stp, ipaH, and aggR) were discarded by PCR. Identification of serotype, phylogenetic group, and antimicrobial resistance was also performed. Results: A total of 41 isolates were characterized. The phylogenetic groups found were B1 in 37 isolates (90.2%), A in 2 (4.8%), and 1 isolate (2.4%) for C and D groups. Serotypes associated with diarrhea in humans (O104:H2 and O154:NM) and hemolytic uremic syndrome (O8:NM) were detected. Thirty-three isolates (80%) were resistant to ceftazidime, 23 (56%), to tetracycline 8 (19.5%) to ampicillin, and 1 to amikacin. Six isolates (14.6%) were multidrug-resistant. Conclusions: This study provides new information about commensal E. coli in slaughtered sheep, high percentages of resistance to antibiotics, and different profiles of antimicrobial resistance were found, their dissemination constitute a risk factor towards the consuming population.

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Microbial and Parasitic Contamination of Fresh Raw Vegetable Samples and Detection of the BlaTEM and BlaCTX-M Genes from E. coli Isolates

Agriculture ◽

10.3390/agriculture10080341 ◽

2020 ◽

Vol 10 (8) ◽

pp. 341

Author(s):

Alina Ghimire ◽

Jitendra Upadhyaya ◽

Tulsi Nayaju ◽

Binod Lekhak ◽

Dhiraj Kumar Chaudhary ◽

...

Keyword(s):

Escherichia Coli ◽

High Rate ◽

Beta Lactamase ◽

Extended Spectrum Beta Lactamase ◽

E Coli ◽

Vegetable Samples ◽

Vegetable Sample ◽

Amoxicillin Clavulanate ◽

Polymerase Chain ◽

Esbl Producers

A total of 100 fresh-raw vegetable samples were collected from the Kathmandu, Lalitpur and Bhaktapur districts of Nepal to evaluate microbial and parasitic contamination, presence of Extended Spectrum Beta Lactamase (ESBL)-producing Escherichia coli and detect the blaTEM and blaCTX-M genes among the Escherichia coli isolates. This study revealed that the prevalence of Giardia cysts was highest (100%) and Hookworm and Entamoeba coli were lowest (1% each). Coliforms were isolated from every raw vegetable sample. A total of 178 bacterial isolates were isolated among which 57 isolates were identified as E. coli, out of which 33 were Multi-drug resistant (MDR) isolates. The high rate of resistance was found towards amoxicillin/clavulanate, tetracycline and cotrimoxazole. The 10 E. coli isolates tested positive in an ESBL screening, out of which 4 were confirmed as ESBL producers by a combined disc test. Out of these 4 confirmed ESBL E. coli, one was found to carry both the blaTEM gene and blaCTX-M genes by the Polymerase Chain Reaction (PCR) technique. One isolate has only the blaTEM gene, while other isolate harboured only blaCTX-M genes.

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Molecular detection of multidrug and colistin-resistant Escherichia coli isolated from house flies in various environmental settings

Future Microbiology ◽

10.2217/fmb-2019-0053 ◽

2019 ◽

Vol 14 (10) ◽

pp. 847-858 ◽

Cited By ~ 7

Author(s):

Abdus Sobur ◽

Zobayda F Haque ◽

Abdullah AM Sabuj ◽

Samina Ievy ◽

AMM Taufiqur Rahman ◽

...

Keyword(s):

Escherichia Coli ◽

Multidrug Resistant ◽

Diffusion Test ◽

Isolation And Identification ◽

Antibiotic Resistant ◽

Disk Diffusion Test ◽

House Flies ◽

E Coli ◽

Pathogenic Escherichia Coli ◽

Antimicrobial Resistance Genes

Aim: To detect antibiotic-resistant pathogenic Escherichia coli in house flies captured from various environmental settings. Materials & methods: Isolation and identification of E. coli was performed based on culture and PCR. Antimicrobial susceptibility testing was conducted using disk diffusion test. Detection of virulence and antimicrobial resistance genes was carried out using PCR methods. Results: The prevalence of E. coli in flies was 61% with highest in dairy farms (70.67%) followed by food centers (65.33%), dustbins (64%) and area close to hospital (44%). The prevalence rates of tetA, tetB, ereA, CITM, SHV, mcr-3 and catA1 resistance determinants were 88.75, 48.52, 41.67, 37.17, 27.77, 18.37 and 14.81%, respectively. Conclusion: Multidrug-resistant pathogenic E. coli are widely distributed in house flies, which requires a one-health approach to delineate their control.

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Prevalence of Extended Spectrum Beta-Lactamase Producing Escherichia coli and Klebsiella species from Urinary Specimens of Children attending Friendship International Children’s Hospital

Nepal Journal of Biotechnology ◽

10.3126/njb.v5i1.18868 ◽

2017 ◽

Vol 5 (1) ◽

pp. 32-38 ◽

Cited By ~ 1

Author(s):

Ujjwal Rimal ◽

Shovana Thapa ◽

Roshani Maharjan

Keyword(s):

Escherichia Coli ◽

Klebsiella Pneumoniae ◽

Beta Lactamase ◽

Disc Diffusion ◽

Extended Spectrum Beta Lactamase ◽

E Coli ◽

Extended Spectrum ◽

Significant Bacteriuria ◽

Klebsiella Spp ◽

Esbl Producers

Extended-spectrum β-lactamase producing E. coli and K. pneumoniae is a serious threat to the patients. These organisms are major extended spectrum beta lactamase (ESBL) producers. The objective of this study was to determine the prevalence of Extended spectrum β- lactamase producing strains of Escherichia coli and Klebsiella spp isolates from the urine sample of children visiting International Friendship Children Hospital. During the seven months, between June 2016 to December 2016, 1018 mid-stream urine samples(MSU) were collected from patients suspected of having UTI. The samples were investigated by conventional semi-quantitative culture technique and identification of E. coli and Klebsiella spp. was done by microscopy and biochemical test. Antibiotic susceptibility test of isolates was performed by modified Kirby Bauer Disc diffusion test. ESBL screening test was done by using 3rd generation Cephalosporin and confirmation done by combination disc diffusion method. Out of total 1018 MSU samples investigated, 200(19.64%) isolates of E. coli and 28(2.7%) isolates of Klebsiella spp. making a total of 228(22.39%) were found to cause significant bacteriuria. 76(33.33%) isolates, from those causing significant bacteriuria, were Multi-drug resistant organisms. Out of 228 isolates, 54(23.68%) were ESBL producers, that includes 51(25.5%) Escherichia coli and 3(12.5%) Klebsiella pneumoniae. ESBL producers were more common in in-patient (36.17%) than out-patient (20.44%). Most of the ESBL producers were resistance to amoxicillin, followed by Cotrimoxazole and Ciprofloxacin respectively. They were highly sensitive to Imipenem, Tigecycline, Amikacin, Piperacillin-Tazobactam, and Nitrofurantoin. High prevalence of ESBL producing E. coli and Klebsiella pneumoniae was found among children. Regular and routine monitoring of ESBL producing isolates is essential.Nepal Journal of Biotechnology. Dec. 2017 Vol. 5, No. 1: 32-38

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Antibiotic-Resistant Escherichia coli and Salmonella from the Feces of Food Animals in the East Province of Rwanda

Animals ◽

10.3390/ani11041013 ◽

2021 ◽

Vol 11 (4) ◽

pp. 1013

Author(s):

Rosine Manishimwe ◽

Paola M. Moncada ◽

Vestine Musanayire ◽

Anselme Shyaka ◽

H. Morgan Scott ◽

...

Keyword(s):

Escherichia Coli ◽

Culture Media ◽

Future Research ◽

Third Generation ◽

Antibiotic Resistant ◽

Disk Diffusion Test ◽

Extended Spectrum Beta Lactamase ◽

E Coli ◽

Food Animals ◽

Third Generation Cephalosporins

In Rwanda, information on antibiotic resistance in food animals is scarce. This study was conducted to detect and phenotypically characterize antibiotic-resistant Escherichia coli and Salmonella in feces of cattle, goats, pigs, and poultry in the East province of Rwanda. We isolated non-type-specific (NTS) E. coli and Salmonella using plain culture media. In addition, we used MacConkey agar media supplemented with cefotaxime at 1.0 μg/mL and ciprofloxacin at 0.5 μg/mL to increase the probability of detecting E. coli with low susceptibility to third-generation cephalosporins and quinolones, respectively. Antibiotic susceptibility testing was performed using the disk diffusion test. Among 540 NTS E. coli isolates, resistance to tetracycline was the most frequently observed (35.6%), followed by resistance to ampicillin (19.6%) and streptomycin (16.5%). Percentages of NTS E. coli resistant to all three antibiotics and percentages of multidrug-resistant strains were higher in isolates from poultry. All isolated Salmonella were susceptible to all antibiotics. The sample-level prevalence for resistance to third-generation cephalosporins was estimated at 35.6% with all third-generation cephalosporin-resistant E. coli, expressing an extended-spectrum beta-lactamase phenotype. The sample-level prevalence for quinolone resistance was estimated at 48.3%. These results provided a baseline for future research and the development of integrated surveillance initiatives.

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