Duplication of blaCTX-M-1 and a class 1 integron on the chromosome enhances antimicrobial resistance in Escherichia coli isolated from racehorses in Japan

Escherichia coli is a serious cause of a variety of hospital-acquired infections and commonly contributes to the environment by house flies. Integrons, particularly class 1 integrons, are the genetic elements that play an important role in the horizontal transfer of antimicrobial resistance mechanism. This mechanism is commonly found in Enterobacteriaceae, especially E. coli. In this study, we aim to investigate the occurrence and antimicrobial resistance patterns of E. coli isolated from the house flies in Phayao hospital and to determine the gene expression of class 1 integrons in those isolates of E. coli. Totally, 70 isolates of E. coli were isolated from 60 house flies collected from the hospital. Fifty-seven of the isolates (81.43 %) were multidrug resistance (MDR) and highly resistant to b-lactams, tetracyclines, and sulfonamides. Of 57 isolates of MDR-E. coli, 20 isolates (35 %) were found to carry class 1 integron genes. Fifteen patterns of antimicrobial resistance occurred in the isolates of integron-positive E. coli. Most integron-positive E. coli isolates were resistant to 7 antimicrobials. Two isolates of these bacteria (10 %) were able to resist 13 out of 14 tested antimicrobials. Using PCR and sequencing analysis, an investigation showed that dfrA17-aadA5, dfrA12-aadA2 gene cassette was the most prevalent cassette (n = 10; 50 %) among the integron-positive E. coli isolates. Our results indicated that the presences of multidrug resistance and class 1 integrons were common in E. coli isolated from the houseflies in hospital. Therefore, screening for integron-positive E. coli from the hospital environment might be necessary for prevention of nosocomial infections.

Download Full-text

Molecular characterisation of antimicrobial resistance and virulence genes in Escherichia coli strains isolated from diarrhoeic and healthy rabbits in Tunisia

World Rabbit Science ◽

10.4995/wrs.2020.10879 ◽

2020 ◽

Vol 28 (2) ◽

pp. 81

Author(s):

Raouia Ben Rhouma ◽

Ahlem Jouini ◽

Amira Klibi ◽

Safa Hamrouni ◽

Aziza Boubaker ◽

...

Keyword(s):

Escherichia Coli ◽

Antimicrobial Resistance ◽

Virulence Genes ◽

Antimicrobial Agents ◽

Antibiotic Resistance Genes ◽

Virulence Gene ◽

Diffusion Method ◽

Class 1 Integron ◽

E Coli ◽

Class 1

The purpose of this study was to identify Escherichia coli isolates in diarrhoeic and healthy rabbits in Tunisia and characterise their virulence and antibiotic resistance genes. In the 2014-2015 period, 60 faecal samples from diarrhoeic and healthy rabbits were collected from different breeding farms in Tunisia. Susceptibility to 14 antimicrobial agents was tested by disc diffusion method and the mechanisms of gene resistance were evaluated using polymerase chain reaction and sequencing methods. Forty E. coli isolates were recovered in selective media. High frequency of resistance to tetracycline (95%) was detected, followed by different levels of resistance to sulphonamide (72.5%), streptomycin (62.5%), trimethoprim-sulfamethoxazole (60%), nalidixic acid (32.5%), ampicillin (37.5%) and ticarcillin (35%). E. coli strains were susceptible to cefotaxime, ceftazidime and imipenem. Different variants of blaTEM, tet, sul genes were detected in most of the strains resistant to ampicillin, tetracycline and sulphonamide, respectively. The presence of class 1 integron was studied in 29 sulphonamide-resistant E. coli strains from which 15 harboured class 1 integron with four different arrangements of gene cassettes, dfrA17+aadA5 (n=9), dfrA1 + aadA1 (n=4), dfrA12 + addA2 (n=1), dfrA12+orf+addA2 (n=1). The qnrB gene was detected in six strains out of 13 quinolone-resistant E. coli strains. Seventeen E. coli isolates from diarrhoeic rabbits harboured the enteropathogenic eae genes associated with different virulence genes tested (fimA, cnf1, aer), and affiliated to B2 (n=8) and D (n=9) phylogroups. Isolated E. coli strains from healthy rabbit were harbouring fim A and/or cnf1 genes and affiliated to A and B1 phylogroups. This study showed that E. coli strains from the intestinal tract of rabbits are resistant to the widely prescribed antibiotics in medicine. Therefore, they constitute a reservoir of antimicrobial-resistant genes, which may play a significant role in the spread of antimicrobial resistance. In addition, the eae virulence gene seemed to be implicated in diarrhoea in breeder rabbits in Tunisia.

Download Full-text

Antimicrobial Resistance, Extended-Spectrum β-Lactamase Productivity, and Class 1 Integrons in Escherichia coli from Healthy Swine

Journal of Food Protection ◽

10.4315/0362-028x.jfp-14-445 ◽

2015 ◽

Vol 78 (8) ◽

pp. 1442-1450 ◽

Cited By ~ 17

Author(s):

KANJANA CHANGKAEW ◽

APIRADEE INTARAPUK ◽

FUANGFA UTRARACHKIJ ◽

CHIE NAKAJIMA ◽

ORASA SUTHIENKUL ◽

...

Keyword(s):

Escherichia Coli ◽

Antimicrobial Resistance ◽

Intestinal Flora ◽

Class 1 Integron ◽

Potential Health ◽

Gene Cassettes ◽

E Coli ◽

Class 1 Integrons ◽

Extended Spectrum ◽

Class 1

Administration of antimicrobials to food-producing animals increases the risk of higher antimicrobial resistance in the normal intestinal flora of these animals. The present cross-sectional study was conducted to investigate antimicrobial susceptibility and extended-spectrum β-lactamase (ESBL)–producing strains and to characterize class 1 integrons in Escherichia coli in healthy swine in Thailand. All 122 of the tested isolates had drug-resistant phenotypes. High resistance was found to ampicillin (98.4% of isolates), chloramphenicol (95.9%), gentamicin (78.7%), streptomycin (77.9%), tetracycline (74.6%), and cefotaxime (72.1%). Fifty-four (44.3%) of the E. coli isolates were confirmed as ESBL-producing strains. Among them, blaCTX-M (45 isolates) and blaTEM (41 isolates) were detected. Of the blaCTX-M-positive E. coli isolates, 37 carried the blaCTX-M-1 cluster, 12 carried the blaCTX-M-9 cluster, and 5 carried both clusters. Sequence analysis revealed blaTEM-1, blaTEM-135, and blaTEM-175 in 38, 2, and 1 isolate, respectively. Eighty-seven (71%) of the 122isolates carried class 1 integrons, and eight distinct drug-resistance gene cassettes with seven different integron profiles were identified in 43 of these isolates. Gene cassettes were associated with resistance to aminoglycosides (aadA1, aadA2, aadA22, or aadA23), trimethoprim (dfrA5, dfrA12, or dfrA17), and lincosamide (linF). Genes encoding β-lactamases were not found in class 1 integrons. This study is the first to report ESBL-producing E. coli with a class 1 integron carrying the linF gene cassette in swine in Thailand. Our findings confirm that swine can be a reservoir of ESBL-producing E. coli harboring class 1 integrons, which may become a potential health risk if these integrons are transmitted to humans. Intensive analyses of animal, human, and environmental isolates are needed to control the spread of ESBL-producing E. coli strains.

Download Full-text

Snapshot Study of Whole Genome Sequences of Escherichia coli from Healthy Companion Animals, Livestock, Wildlife, Humans and Food in Italy

Antibiotics ◽

10.3390/antibiotics9110782 ◽

2020 ◽

Vol 9 (11) ◽

pp. 782

Author(s):

Elisa Massella ◽

Cameron J. Reid ◽

Max L. Cummins ◽

Kay Anantanawat ◽

Tiziana Zingali ◽

...

Keyword(s):

Escherichia Coli ◽

Antimicrobial Resistance ◽

Companion Animals ◽

Poultry Meat ◽

Whole Genome ◽

Class 1 Integron ◽

Antimicrobial Resistance Genes ◽

Class 1 ◽

Insertion Elements ◽

Chromosomal Mutations

Animals, humans and food are all interconnected sources of antimicrobial resistance (AMR), allowing extensive and rapid exchange of AMR bacteria and genes. Whole genome sequencing (WGS) was used to characterize 279 Escherichia coli isolates obtained from animals (livestock, companion animals, wildlife), food and humans in Italy. E. coli predominantly belonged to commensal phylogroups B1 (46.6%) and A (29%) using the original Clermont criteria. One hundred and thirty-six sequence types (STs) were observed, including different pandemic (ST69, ST95, ST131) and emerging (ST10, ST23, ST58, ST117, ST405, ST648) extraintestinal pathogenic Escherichia coli (ExPEC) lineages. Eight antimicrobial resistance genes (ARGs) and five chromosomal mutations conferring resistance to highest priority critically important antimicrobials (HP-CIAs) were identified (qnrS1, qnrB19, mcr-1, blaCTX-M1,15,55, blaCMY-2, gyrA/parC/parE, ampC and pmrB). Twenty-two class 1 integron arrangements in 34 strains were characterized and 11 ARGs were designated as intI1 related gene cassettes (aadA1, aadA2, aadA5, aad23, ant2_Ia, dfrA1, dfrA7, dfrA14, dfrA12, dfrA17, cmlA1). Notably, most intI1 positive strains belonged to rabbit (38%) and poultry (24%) sources. Three rabbit samples carried the mcr-1 colistin resistance gene in association with IS6 family insertion elements. Poultry meat harbored some of the most prominent ExPEC STs, including ST131, ST69, ST10, ST23, and ST117. Wildlife showed a high average number of virulence-associated genes (VAGs) (mean = 10), mostly associated with an ExPEC pathotype and some predominant ExPEC lineages (ST23, ST117, ST648) were identified.

Download Full-text

Antimicrobial Resistance in Class 1 Integron-Positive Shiga Toxin-Producing Escherichia coli Isolated from Cattle, Pigs, Food and Farm Environment

Microorganisms ◽

10.3390/microorganisms6040099 ◽

2018 ◽

Vol 6 (4) ◽

pp. 99 ◽

Cited By ~ 2

Author(s):

Rocío Colello ◽

Alejandra Krüger ◽

José Conza ◽

John Rossen ◽

Alexander Friedrich ◽

...

Keyword(s):

Escherichia Coli ◽

Antimicrobial Resistance ◽

Shiga Toxin ◽

Variable Region ◽

Diffusion Method ◽

Class 1 Integron ◽

Integrase Gene ◽

Class 1 Integrons ◽

Antimicrobial Resistance Genes ◽

Class 1

The aim of this study was to investigate the presence of class 1 integrons in a collection of Shiga toxin-producing Escherichia coli (STEC) from different origins and to characterize pheno- and genotypically the antimicrobial resistance associated to them. A collection of 649 isolates were screened for the class 1 integrase gene (intI1) by Polymerase chain reaction The variable region of class 1 integrons was amplified and sequenced. Positive strains were evaluated for the presence of antimicrobial resistance genes with microarray and for antimicrobial susceptibility by the disk diffusion method. Seven out of 649 STEC strains some to serogroups, O26, O103 and O130 isolated from cattle, chicken burger, farm environment and pigs were identified as positive for intl1. Different arrangements of gene cassettes were detected in the variable region of class 1 integron: dfrA16, aadA23 and dfrA1-aadA1. In almost all strains, phenotypic resistance to streptomycin, tetracycline, trimethoprim/sulfamethoxazole, and sulfisoxazole was observed. Microarray analyses showed that most of the isolates carried four or more antimicrobial resistance markers and STEC strains were categorized as Multridrug-resistant. Although antimicrobials are not usually used in the treatment of STEC infections, the presence of Multridrug-resistant in isolates collected from farm and food represents a risk for animal and human health.

Download Full-text

Truncated Class 1 Integron Gene Cassette Arrays Contribute to Antimicrobial Resistance of Diarrheagenic Escherichia coli

BioMed Research International ◽

10.1155/2020/4908189 ◽

2020 ◽

Vol 2020 ◽

pp. 1-9

Author(s):

Akiko Kubomura ◽

Tsuyoshi Sekizuka ◽

Daisuke Onozuka ◽

Koichi Murakami ◽

Hirokazu Kimura ◽

...

Keyword(s):

Escherichia Coli ◽

Antimicrobial Resistance ◽

Gene Cassette ◽

Class 1 Integron ◽

Gene Cassettes ◽

E Coli ◽

Antimicrobial Resistance Genes ◽

Class 1 ◽

Stool Samples ◽

Human Stool

Class 1 integrons (c1-integrons) are associated with multidrug resistance in diarrheagenic Escherichia coli (DEC). However, little is known about gene cassettes located within these c1-integrons, particularly truncated c1-integrons, in DEC strains. Therefore, the aims of the present study were to reveal the relationship between antimicrobial resistance and the presence of truncated c1-integrons in DEC isolates derived from human stool samples in Japan. A total of 162 human stool-derived DEC isolates from Japan were examined by antimicrobial susceptibility testing, PCR-based gene detection, and next-generation sequencing analyses. Results showed that 44.4% (12/27) of c1-integrons identified in the DEC isolates harbored only intI1 (an element of c1-integrons) and were truncated by IS26, Tn3, or IS1-group insertion sequences. No difference in the frequency of antimicrobial resistance was recorded between intact and truncated c1-integron-positive DEC isolates. Isolates containing intact/truncated c1-integrons, particularly enteroaggregative E. coli isolates, were resistant to a greater number of antimicrobials than isolates without c1-integrons. aadA and dfrA were the most prevalent antimicrobial resistance genes in the intact/truncated c1-integrons examined in this study. Therefore, gene cassettes located within these intact/truncated c1-integrons may only play a limited role in conferring antimicrobial resistance among DEC. However, DEC harboring truncated c1-integrons may be resistant to a greater number of antimicrobials than c1-integron-negative DEC, similar to strains harboring intact c1-integrons.

Download Full-text

Antimicrobial Resistance and Presence of Class 1 Integrons Among Different Serotypes of Salmonella spp. Recovered From Children with Diarrhea in Tehran, Iran

Infectious Disorders - Drug Targets ◽

10.2174/1871526519666190130171020 ◽

2020 ◽

Vol 20 (2) ◽

pp. 160-166

Author(s):

Seyedeh Hanieh Eshaghi Zadeh ◽

Hossein Fahimi ◽

Fatemeh Fardsanei ◽

Mohammad Mehdi Soltan Dallal

Keyword(s):

Antibiotic Resistance ◽

Antimicrobial Resistance ◽

Multidrug Resistant ◽

Salmonella Spp ◽

Class 1 Integron ◽

Class 1 Integrons ◽

Food Borne ◽

Class 1 ◽

Resistance Patterns ◽

C 30

Background: Salmonellosis is a major food-borne disease worldwide. The increasing prevalence of antimicrobial resistance among food-borne pathogens such as Salmonella spp. is concerning. Objective: The main objective of this study is to identify class 1 integron genes and to determine antibiotic resistance patterns among Salmonella isolates from children with diarrhea. Methods: A total of 30 Salmonella isolates were recovered from children with diarrhea. The isolates were characterized for antimicrobial susceptibility and screened for the presence of class 1 integron genes (i.e. intI1, sulI1, and qacEΔ1). Results: The most prevalent serotype was Enteritidis 36.7%, followed by Paratyphi C (30%), and Typhimurium (16.7%). The highest rates of antibiotic resistance were obtained for nalidixic acid (53.3%), followed by streptomycin (40%), and tetracycline (36.7%). Regarding class 1 integrons, 36.7%, 26.7%, and 33.3% of the isolates carried intI1, SulI, and qacEΔ1, respectively, most of which (81.8%) were multidrug-resistant (MDR). Statistical analysis revealed that the presence of class 1 integron was significantly associated with resistance to streptomycin and tetracycline (p = 0.042). However, there was no association between class 1 integron and other antibiotics used in this study (p > 0.05). Conclusion: The high frequency of integron class 1 gene in MDR Salmonella strains indicates that these mobile genetic elements are versatile among different Salmonella serotypes, and associated with reduced susceptibility to many antimicrobials.

Download Full-text

Impact of Feed Supplementation with Antimicrobial Agents on Growth Performance of Broiler Chickens, Clostridium perfringens and Enterococcus Counts, and Antibiotic Resistance Phenotypes and Distribution of Antimicrobial Resistance Determinants in Escherichia coli Isolates

Applied and Environmental Microbiology ◽

10.1128/aem.01086-07 ◽

2007 ◽

Vol 73 (20) ◽

pp. 6566-6576 ◽

Cited By ~ 103

Author(s):

Moussa S. Diarra ◽

Fred G. Silversides ◽

Fatoumata Diarrassouba ◽

Jane Pritchard ◽

Luke Masson ◽

...

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Antimicrobial Resistance ◽

Broiler Chickens ◽

Antimicrobial Agents ◽

Growth Promoters ◽

Feed Supplementation ◽

E Coli ◽

Resistance Determinants ◽

Class 1

ABSTRACT The effects of feed supplementation with the approved antimicrobial agents bambermycin, penicillin, salinomycin, and bacitracin or a combination of salinomycin plus bacitracin were evaluated for the incidence and distribution of antibiotic resistance in 197 commensal Escherichia coli isolates from broiler chickens over 35 days. All isolates showed some degree of multiple antibiotic resistance. Resistance to tetracycline (68.5%), amoxicillin (61.4%), ceftiofur (51.3%), spectinomycin (47.2%), and sulfonamides (42%) was most frequent. The levels of resistance to streptomycin, chloramphenicol, and gentamicin were 33.5, 35.5, and 25.3%, respectively. The overall resistance levels decreased from day 7 to day 35 (P < 0.001). Comparing treatments, the levels of resistance to ceftiofur, spectinomycin, and gentamicin (except for resistance to bacitracin treatment) were significantly higher in isolates from chickens receiving feed supplemented with salinomycin than from the other feeds (P < 0.001). Using a DNA microarray analysis capable of detecting commonly found antimicrobial resistance genes, we characterized 104 tetracycline-resistant E. coli isolates from 7- to 28-day-old chickens fed different growth promoters. Results showed a decrease in the incidence of isolates harboring tet(B), bla TEM, sulI, and aadA and class 1 integron from days 7 to 35 (P < 0.01). Of the 84 tetracycline-ceftiofur-resistant E. coli isolates, 76 (90.5%) were positive for bla CMY-2. The proportions of isolates positive for sulI, aadA, and integron class 1 were significantly higher in salinomycin-treated chickens than in the control or other treatment groups (P < 0.05). These data demonstrate that multiantibiotic-resistant E. coli isolates can be found in broiler chickens regardless of the antimicrobial growth promoters used. However, the phenotype and the distribution of resistance determinants in E. coli can be modulated by feed supplementation with some of the antimicrobial agents used in broiler chicken production.

Download Full-text

Characterization of a Novel Plasmid-Mediated Cephalosporinase (CMY-9) and Its Genetic Environment in an Escherichia coli Clinical Isolate

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.46.8.2427-2434.2002 ◽

2002 ◽

Vol 46 (8) ◽

pp. 2427-2434 ◽

Cited By ~ 41

Author(s):

Yohei Doi ◽

Naohiro Shibata ◽

Keigo Shibayama ◽

Kazunari Kamachi ◽

Hiroshi Kurokawa ◽

...

Keyword(s):

Escherichia Coli ◽

Amino Acid ◽

Amino Acid Sequence ◽

Gene Cassette ◽

Single Amino Acid ◽

Common Region ◽

Class 1 Integron ◽

Class 1 ◽

The Common ◽

High Level

ABSTRACT An Escherichia coli strain, HKYM68, which showed resistance to broad-spectrum cephalosporins was isolated from a sputum specimen in Japan. The high-level resistance of the strain to ceftazidime, cefpirome, and moxalactam was carried by a self-transferable plasmid. The β-lactamase gene responsible for the resistance was cloned and sequenced. The deduced amino acid sequence of this gene product, CMY-9, had a single amino acid substitution (E85D), the residue reported to be part of the recognition site for the R1 side chain of β-lactams, compared with the amino acid sequence of CMY-8 and also had 78% identity with the amino acid sequence of CepH, a chromosomal cephalosporinase of Aeromonas hydrophila. A sul1-type class 1 integron containing an aacA1-orfG gene cassette was identified upstream of bla CMY-9 and ended with a truncated 3′ conserved segment. The following 2.1 kb was almost identical to the common region of integrons In6 and In7 and the integron of pSAL-1, except that orf513 encoding a putative transposase was identified instead of orf341 due to addition of a single nucleotide. bla CMY-9 was closely located downstream of the end of the common region. These observations are indicative of the exogenous derivation of bla CMY-9 from some environmental microorganisms such as aeromonads.

Download Full-text