Molecular Detection of Colistin Resistance mcr-1 Gene in Multidrug-Resistant Escherichia coli Isolated from Chicken

Zoonotic and antimicrobial-resistant Escherichia coli (hereafter, E. coli) is a global public health threat which can lead to detrimental effects on human health. Here, we aim to investigate the antimicrobial resistance and the presence of mcr-1 gene in E. coli isolated from chicken feces. Ninety-four E. coli isolates were obtained from samples collected from different locations in Bangladesh, and the isolates were identified using conventional microbiological tests. Phenotypic disk diffusion tests using 20 antimicrobial agents were performed according to CLSI-EUCAST guidelines, and minimum inhibitory concentrations (MICs) were determined for a subset of samples. E. coli isolates showed high resistance to colistin (88.30%), ciprofloxacin (77.66%), trimethoprim/sulfamethoxazole (76.60%), tigecycline (75.53%), and enrofloxacin (71.28%). Additionally, the pathotype eaeA gene was confirmed in ten randomly selected E. coli isolates using primer-specific polymerase chain reaction (PCR). The presence of mcr-1 gene was confirmed using PCR and sequencing analysis in six out of ten E. coli isolates. Furthermore, sequencing and phylogenetic analyses revealed a similarity between the catalytic domain of Neisseria meningitidis lipooligosaccharide phosphoethanolamine transferase A (LptA) and MCR proteins, indicating that the six tested isolates were colistin resistant. Finally, the findings of the present study showed that E. coli isolated from chicken harbored mcr-1 gene, and multidrug and colistin resistance. These findings accentuate the need to implement strict measures to limit the imprudent use of antibiotics, particularly colistin, in agriculture and poultry farms.

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Emergence of colistin resistance in multidrug-resistant Klebsiella pneumoniae and Escherichia coli strains isolated from cancer patients

Annals of Clinical Microbiology and Antimicrobials ◽

10.1186/s12941-019-0339-4 ◽

2019 ◽

Vol 18 (1) ◽

Cited By ~ 8

Author(s):

Mai M. Zafer ◽

Hadir A. El-Mahallawy ◽

Asmaa Abdulhak ◽

Magdy A. Amin ◽

Mohamed H. Al-Agamy ◽

...

Keyword(s):

Escherichia Coli ◽

Klebsiella Pneumoniae ◽

Cancer Patients ◽

Clonal Diversity ◽

Multidrug Resistant ◽

Resistance Mechanisms ◽

Global Public Health ◽

Colistin Resistance ◽

E Coli ◽

Recent Emergence

Abstract Background Colistin resistance is mainly driven by alterations in the Gram-negative outer membrane lipopolysaccharides and is caused, in most cases, by mutations in mgrB gene. However, the recent emergence of plasmid-encoded colistin resistance among Enterobacteriaceae strains represents a serious threat to global public health. In this paper we have investigated the rates of colistin resistance and the underlying mechanisms in 450 Klebsiella pneumoniae and Escherichia coli isolates obtained from cancer patients in Egypt. Methods Colistin susceptibility and minimum inhibitory concentrations were determined according to the European Committee on Antimicrobial Susceptibility Testing, by broth microdilution, and by E-test. The mcr-1, mcr-2 and mgrB genes were detected by PCR and then sequenced. Clonal diversity in colistin-resistant K. pneumoniae was evaluated by multilocus sequence typing. Results Forty (8.8%) colistin-resistant isolates, including 22 K. pneumoniae and 18 E. coli, were isolated over 18 months. Of these, 50% were carbapenem-resistant, out of which nine were blaOXA-48 and seven blaNDM-1 positive. The mechanisms of colistin resistance could be revealed only in three of the 40 resistant strains, being represented by mcr-1 in one blaNDM-1-positive E. coli strain and in one K. pneumoniae ST11 and by mgrB mutations, detected in one K. pneumoniae isolate. None of the studied isolates harbored mcr-2. Conclusions Our results demonstrate a high frequency of colistin resistance in enterobacterial strains isolated from cancer patients, but a low prevalence of the most well known resistance mechanisms.

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Molecular survey of mcr1 and mcr2 plasmid mediated colistin resistance genes in Escherichia coli isolates of animal origin in Iran

BMC Research Notes ◽

10.1186/s13104-021-05519-6 ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

Kayhan Ilbeigi ◽

Mahdi Askari Badouei ◽

Hossein Vaezi ◽

Hassan Zaheri ◽

Sina Aghasharif ◽

...

Keyword(s):

Escherichia Coli ◽

Resistance Genes ◽

Companion Animals ◽

Multidrug Resistant ◽

Animal Origin ◽

Colistin Resistance ◽

E Coli ◽

High Level ◽

Farming Industry

Abstract Objectives The emergence of colistin-resistant Enterobacteriaceae from human and animal sources is one of the major public health concerns as colistin is the last-resort antibiotic for treating infections caused by multidrug-resistant Gram-negative bacteria. We aimed to determine the prevalence of the prototype widespread colistin resistance genes (mcr-1 and mcr-2) among commensal and pathogenic Escherichia coli strains isolated from food-producing and companion animals in Iran. Results A total of 607 E. coli isolates which were previously collected from different animal sources between 2008 and 2016 used to uncover the possible presence of plasmid-mediated colistin resistance genes (mcr-1 and mcr-2) by PCR. Overall, our results could not confirm the presence of any mcr-1 or mcr-2 positive E. coli among the studied isolates. It is concluded that despite the important role of food-producing animals in transferring the antibiotic resistance, they were not the main source for carriage of mcr-1 and mcr-2 in Iran until 2016. This study suggests that the other mcr variants (mcr-3 to mcr-9) might be responsible for conferring colistin resistance in animal isolates in Iran. The possible linkage between pig farming industry and high level of mcr carriage in some countries needs to be clarified in future prospective studies.

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Heterogeneous and Flexible Transmission ofmcr-1in Hospital-AssociatedEscherichia coli

mBio ◽

10.1128/mbio.00943-18 ◽

2018 ◽

Vol 9 (4) ◽

Cited By ~ 23

Author(s):

Yingbo Shen ◽

Zuowei Wu ◽

Yang Wang ◽

Rong Zhang ◽

Hong-Wei Zhou ◽

...

Keyword(s):

Escherichia Coli ◽

Genomic Analysis ◽

Multidrug Resistant ◽

Fluoroquinolone Resistance ◽

Gram Negative Bacteria ◽

Colistin Resistance ◽

Gram Negative ◽

Content Type ◽

E Coli ◽

Genes Encoding

ABSTRACTThe recent emergence of a transferable colistin resistance mechanism, MCR-1, has gained global attention because of its threat to clinical treatment of infections caused by multidrug-resistant Gram-negative bacteria. However, the possible transmission route ofmcr-1amongEnterobacteriaceaespecies in clinical settings is largely unknown. Here, we present a comprehensive genomic analysis ofEscherichia coliisolates collected in a hospital in Hangzhou, China. We found thatmcr-1-carrying isolates from clinical infections and feces of inpatients and healthy volunteers were genetically diverse and were not closely related phylogenetically, suggesting that clonal expansion is not involved in the spread ofmcr-1. Themcr-1gene was found on either chromosomes or plasmids, but in most of theE. coliisolates,mcr-1was carried on plasmids. The genetic context of the plasmids showed considerable diversity as evidenced by the different functional insertion sequence (IS) elements, toxin-antitoxin (TA) systems, heavy metal resistance determinants, and Rep proteins of broad-host-range plasmids. Additionally, the genomic analysis revealed nosocomial transmission ofmcr-1and the coexistence ofmcr-1with other genes encoding β-lactamases and fluoroquinolone resistance in theE. coliisolates. These findings indicate thatmcr-1is heterogeneously disseminated in both commensal and pathogenic strains ofE. coli, suggest the high flexibility of this gene in its association with diverse genetic backgrounds of the hosts, and provide new insights into the genome epidemiology ofmcr-1among hospital-associatedE. colistrains.IMPORTANCEColistin represents one of the very few available drugs for treating infections caused by extensively multidrug-resistant Gram-negative bacteria. The recently emergentmcr-1colistin resistance gene threatens the clinical utility of colistin and has gained global attention. Howmcr-1spreads in hospital settings remains unknown and was investigated by whole-genome sequencing ofmcr-1-carryingEscherichia coliin this study. The findings revealed extraordinary flexibility ofmcr-1in its spread among genetically diverseE. colihosts and plasmids, nosocomial transmission ofmcr-1-carryingE. coli, and the continuous emergence of novel Inc types of plasmids carryingmcr-1and newmcr-1variants. Additionally,mcr-1was found to be frequently associated with other genes encoding β-lactams and fluoroquinolone resistance. These findings provide important information on the transmission and epidemiology ofmcr-1and are of significant public health importance as the information is expected to facilitate the control of this significant antibiotic resistance threat.

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T4-Like Phage Bp7, a Potential Antimicrobial Agent for Controlling Drug-Resistant Escherichia coli in Chickens

Applied and Environmental Microbiology ◽

10.1128/aem.01505-13 ◽

2013 ◽

Vol 79 (18) ◽

pp. 5559-5565 ◽

Cited By ~ 21

Author(s):

Can Zhang ◽

Wenli Li ◽

Wenhua Liu ◽

Ling Zou ◽

Chen Yan ◽

...

Keyword(s):

Escherichia Coli ◽

Burst Size ◽

Multidrug Resistant ◽

Open Reading Frames ◽

Attractive Alternative ◽

Drug Resistant ◽

Animal Cells ◽

Content Type ◽

E Coli ◽

Chicken Feces

ABSTRACTChicken-pathogenicEscherichia coliis severely endangering the poultry industry in China and worldwide, and antibiotic therapy is facing an increasing problem of antibiotic resistance. Bacteriophages can kill bacteria with no known activity in human or animal cells, making them an attractive alternative to antibiotics. In this study, we present the characteristics of a novel virulent bacteriophage, Bp7, specifically infecting pathogenic multidrug-resistantE. coli. Phage Bp7 was isolated from chicken feces. Bp7 belongs to the familyMyoviridae, possessing an elongated icosahedral head and contractile sheathed tail. It has a 168-kb double-stranded DNA genome. For larger yields, its optimal multiplicity of infection (MOI) to infectE. coliwas about 0.001. The latent period was 10 to 15 min, and the burst size was 90 PFU/infected cell. It was stable both at pH 5.0 to 10.0 and at 40°C or 50°C for at least 1 h. Bp7 could infect 46% of pathogenic clinicalE. colistrains. Bp7 harbored 791 open reading frames (ORFs) and 263 possible genes. Among the 263 genes, 199 possessed amino acid sequence identities with ORFs of phage T4, 62 had identities with other T4-like phages, and only one lacked any database match. The genome of Bp7 manifested obvious division and rearrangement compared to phages T4, JS98, and IME08. Bp7 is a new member of the “T4-like” genus, familyMyoviridae. Its wide host range, strong cell-killing activity, and high stability to pH make it an alternative to antimicrobials for controlling drug-resistantE. coliin chickens.

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Occurrence of Multi-Drug-Resistant Escherichia coli in Chickens, Humans, Rodents and Household Soil in Karatu, Northern Tanzania

Antibiotics ◽

10.3390/antibiotics10091137 ◽

2021 ◽

Vol 10 (9) ◽

pp. 1137

Author(s):

Valery S. Sonola ◽

Abdul S. Katakweba ◽

Gerald Misinzo ◽

Mecky I. N. Matee

Keyword(s):

Escherichia Coli ◽

Antimicrobial Agents ◽

Multidrug Resistant ◽

Soil Samples ◽

Control Measures ◽

Rodent Control ◽

E Coli ◽

Northern Tanzania ◽

Amoxicillin Clavulanate ◽

Resistance Rates

We investigated antibiotic resistance profiles of Escherichia coli among 960 samples obtained from chickens (236), humans (243), rodents (101) and soil (290). E. coli was isolated from 650 (67.7%) samples. Isolation frequency varied significantly between chickens, humans, rodents and soil samples, being 81.6%, 86.5%, 79.2% and 31.0%, respectively (p < 0.001). Resistance rates were particularly higher against imipenem (79.8%), cefotaxime (79.7%) and tetracycline (73.7%) and moderate against amoxicillin-clavulanate (49.4%). Overall, 78.8% of the isolates were multidrug-resistant (MDR) among which, 38.8%, 25.1%, 12.9% and 2.5% exhibited resistance to three, four, five and six different classes of antibiotics, respectively. Multidrug-resistant E. coli were observed in 27.7%, 30.3%, 10.8% and 10.0% of the isolates from chickens, humans, rodents and soil samples, respectively. Our results show high levels of antimicrobial resistance including MDR in E. coli isolated from chickens, humans, rodents and soil samples in Karatu, Northern Tanzania. Comprehensive interventions using a one-health approach are needed and should include improving (i) awareness of the community on judicious use of antimicrobial agents in humans and animals, (ii) house conditions and waste management and (iii) rodent control measures.

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Emergence of high multidrug-resistant Escherichia coli isolates from neonates with infection

10.21203/rs.3.rs-20364/v1 ◽

2020 ◽

Author(s):

Dan Wu ◽

Yijun Ding ◽

Jinjing Zhang ◽

Kaihu Yao ◽

Wei Gao ◽

...

Keyword(s):

Escherichia Coli ◽

Multidrug Resistance ◽

Clavulanic Acid ◽

Antimicrobial Agents ◽

Multidrug Resistant ◽

Resistant Bacteria ◽

E Coli ◽

Mueller Hinton ◽

Mueller Hinton Agar ◽

Amoxicillin Clavulanic Acid

Abstract Background Escherichia coli (E.coli) rank one of the most common pathogens that can cause neonatal infections. The emergence of antibiotic-resistant bacteria is a major cause of treatment failure in newborn with infection. The purpose of this study was to describe the antibiotic resistance and multidrug-resistance of E.coli isolated from neonates with infection.Methods The antimicrobial susceptibility testing of the E. coli strains to selected antibiotics was assessed with the E-test technique on the Mueller-Hinton agar. The antimicrobials tests were included ceftazidime, cefuroxime, cefatriaxone, amoxicillin, amoxicillin-clavulanic acid, cefoperazone - sulbactam, meropenem, gentamicin, ciprofloxacin and sulfonamides. The minimal inhibitory concerntration (MIC) values of the antimicrobial agents selected for this study was determined by an agar dilution technique on Mueller-Hinton agar according to the Clinical and Laboratory Standards Institute recommendations. Results A total of 100 E. coli strains was isolated from phlegm (n = 78), blood (n = 10), cerebrospinal fluid (n = 5), and umbilical discharge (n = 7) of neonates hospitalized at Beijing Children’s Hospital. The highest resistance rate of E.coli was found in amoxicillin at 85%, followed by cefuroxime 65%, and cefatriaxone 60%, respectively. 6% and 5% of all isolates were resistant to amoxicillin/clavulanic acid and cefoperazone -sulbactam merely. The resistance rates to ceftazidime, gentamicin, ciprofloxacin and sulfonamides were 31%, 20%, 33%, 47%, respectively. All the isolates were susceptible to meropenem. Multidrug resistance was defined in E.coli as resistance to at least three antibiotic families. About 26% (26/100) of all the E.coli isolates were multidrug-resistant. The detection rate of ESBL-Producing E. coli was 55%. The rate in E. coli isolates from phlegm was higher than aseptic humoral. The difference was statistically significant (P < 0.05). It is worth noting that the majority of the isolates were also resistant to non-β-lactam antimicrobial agents, but the resistant rates were significantly lower than extended-spectrum β-lactamases.Conclusions: Multi-drug-resistant E.coli has become a thorny problem in clinical treatment. It is necessary to monitor E. coli resistance.

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Characterization of auto-agglutinating and non-typeable uropathogenic Escherichia coli strains

The Journal of Infection in Developing Countries ◽

10.3855/jidc.11098 ◽

2019 ◽

Vol 13 (06) ◽

pp. 465-472

Author(s):

Ulises Hernández-Chiñas ◽

Alejandro Pérez-Ramos ◽

Laura Belmont-Monroy ◽

María E Chávez-Berrocal ◽

Edgar González-Villalobos ◽

...

Keyword(s):

Escherichia Coli ◽

Virulence Genes ◽

Antimicrobial Agents ◽

Multidrug Resistant ◽

Uropathogenic Escherichia Coli ◽

E Coli ◽

Resistant Strains ◽

Tract Infections ◽

Disk Method

Introduction: Uropathogenic Escherichia coli (UPEC) are the main etiological agent of urinary tract infections (UTIs). Association between different serotypes and UTIs is known, however, some strains are incapable to be serotyped. The aim of this work was to study bthe phenotypical and genotypical characteristics of 113 non-typeable (NT) and auto-agglutinating (AA) E. coli strains, isolated from UTIs in children and adults. Methodology: The 113 UPEC strains were analyzed by PCR assays using specific primers to determine their serogroups, fimH, papC, iutA, sat, hlyCA and cnf1, virulence associated genes, and chuA, yjaA and TSPE4.C2 for phylogroup determination. Additionally, the diffusion disk method was performed to evaluate the antimicrobial resistance to 18 antimicrobial agents. Results: Using the PCR assay, 63% (71) of the strains were genotyped showing O25 and O75 as the most common serogroups. The virulence genes fimH (86%) and iutA (74%) were the most prevalent, in relation to the phylogroups the commensal (A and B1) and virulent (B2 and D) showed similar frequencies (P > 0.05). The antimicrobial susceptibility test showed a high percentage (73%) of multidrug-resistant strains. Conclusions: The genotyping allowed identifying the serogroup in many of the strains that could not be typed by traditional serology. The strains carried virulence genes and were multidrug-resistant in both, commensal and virulent phylogroups. Our findings revealed that, in addition to the classical UPEC serogroups, there are pathogenic serogroups not reported yet.

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611. Fosfomycin Resistance of Multidrug-Resistant Escherichia coli and Mechanisms of Fosfomycin Resistance

Open Forum Infectious Diseases ◽

10.1093/ofid/ofz360.680 ◽

2019 ◽

Vol 6 (Supplement_2) ◽

pp. S285-S285

Author(s):

Hyeri Seok ◽

Ji Hoon Jeon ◽

Hee Kyoung Choi ◽

Won Suk Choi ◽

Dae Won Park ◽

...

Keyword(s):

Escherichia Coli ◽

Antimicrobial Agents ◽

Multidrug Resistant ◽

Resistance Rate ◽

Coli Strain ◽

Resistant Bacteria ◽

E Coli ◽

Broth Microdilution Method ◽

Fosfomycin Resistance

Abstract Background Fosfomycin is one of the antibiotics that may be a candidate for the next-generation antimicrobial agents againt multidrug-resistant bacteria. To date, it is known that the resistance rate is not high for Escherichia coli. However, it is necessary to update the fosfomycin resistance rates in E. coli according to the studies that extended spectrum β-lactamase (ESBL) producing E. coli strains are highly resistance to fosfomycin. We evaluated the resistance rate of fosfomycin, the resistant mechanism of fosfomycin in E. coli, and the activity of fosfomycin against susceptible and resistant strains of E. coli. Methods A total of 283 clinical isolates was collected from patients with Escherichia coli species during the period of January 2018 to June 2018, in three tertiary hospitals of Republic of Korea. In vitro antimicrobial susceptibility tests were performed in all E. coli isolates using the broth microdilution method according to the Clinical and Laboratory Standard Institute (CLSI). Multilocus sequence typing (MLST) of the Oxford scheme was conducted to determine the genotypes of E. coli isolated. Fosfomycin genes were investigated for all fosfomycin-resistant E. coli strains. Results The overall resistance rate to fosfomycin was 10.2%, compared with 53.4%, 46.3%, 41.3%, 31.1%, 10.6%, 2.5%, and 2.1% for ciprofloxacin, cefixime, cefepime, piperacillin/tazobactam, colistin, ertapenem, and amikacin, respectively. The 29 fosfomycin-resistant isolates did not show a clonal pattern on the phylogenetic tree. MurA and glp genes were identified in all strains. FosA3 were identified in two strains and uhp gene were identified in 4 strains. In time-kill curve studies, fosfomycin was more bactericidal than cefixime against all sensitive E. coli strain. Morever, fosfomycin was more bactericidal than piperacillin/tazobactam against ESBL-producing E. coli strain. Conclusion The resistant rate of fosfomycin to E. coli is still low. Fosfomycin was active against E. coli including ESBL producing strains. Disclosures All authors: No reported disclosures.

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Activity of Tigecycline or Colistin in Combination with Zidovudine against Escherichia coli Harboring tet(X) and mcr-1

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01172-20 ◽

2020 ◽

Vol 65 (1) ◽

pp. e01172-20 ◽

Cited By ~ 1

Author(s):

Yu-Feng Zhou ◽

Ping Liu ◽

Shu-He Dai ◽

Jian Sun ◽

Ya-Hong Liu ◽

...

Keyword(s):

Escherichia Coli ◽

Multidrug Resistant ◽

Therapeutic Options ◽

Infection Model ◽

Synergistic Activity ◽

Colistin Resistance ◽

Content Type ◽

E Coli ◽

Antiretroviral Drug

ABSTRACTAlternative therapeutic options are urgently needed against multidrug-resistant Escherichia coli infections, especially in situations of preexisting tigecycline and colistin resistance. Here, we investigated synergistic activity of the antiretroviral drug zidovudine in combination with tigecycline or colistin against E. coli harboring tet(X) and mcr-1 in vitro and in a murine thigh infection model. Zidovudine and tigecycline/colistin combinations achieved synergistic killing and significantly decreased bacterial burdens by >2.5-log10 CFU/g in thigh tissues compared to each monotherapy.

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Characterization of multidrug-resistant Escherichia coli isolated from extraintestinal clinical infections in animals

Journal of Medical Microbiology ◽

10.1099/jmm.0.018002-0 ◽

2010 ◽

Vol 59 (5) ◽

pp. 592-598 ◽

Cited By ~ 19

Author(s):

Justine S. Gibson ◽

Rowland N. Cobbold ◽

Darren J. Trott

Keyword(s):

Escherichia Coli ◽

Antimicrobial Susceptibility ◽

Animal Species ◽

Antimicrobial Agents ◽

Multidrug Resistant ◽

Phylogenetic Group ◽

Phylogenetic Groups ◽

Diagnostic Laboratory ◽

E Coli

Multidrug-resistant (MDR) Escherichia coli causes extraintestinal infections in both humans and animals. This study aimed to determine whether MDR E. coli isolates cultured from extraintestinal infections in several animal species were clonal and crossed host-species boundaries, as suggested by initial characterization of a subset of canine and human isolates, or whether they represented a diverse group of host-specific strains. Isolates were obtained either from The University of Queensland Veterinary Diagnostic Laboratory or from an independent diagnostic laboratory between October 1999 and December 2007. Ninety-six MDR E. coli isolates cultured from extraintestinal clinical infections in 55 animals comprising dogs (n=45), cats (n=5), horses (n=4) and a koala (n=1) were analysed by phylogenetic grouping, antimicrobial susceptibility testing and PFGE. The isolates were cultured from the urinary tract (n=61), reproductive tract (n=11), wounds (n=11), surgical site infections (n=4) and other sites (n=9). Isolates from the same E. coli phylogenetic group with 100 % PFGE similarity and the same antimicrobial susceptibility pattern were considered to be repeat clones and excluded from further analysis. Three of the four E. coli phylogenetic groups (A, n=19; B1, n=8; and D, n=49) were represented. Analysis of PFGE similarity identified clusters of related phylogenetic group A isolates [clonal group (CG) 1] and group D isolates (CG2 and CG3), with the remainder of the isolates demonstrating diversity. The majority of CG2 isolates contained a plasmid-borne AmpC β-lactamase, imparting resistance to cefoxitin and third-generation cephalosporins, and were obtained between 2000 and 2003. CG3 isolates were sensitive to these antimicrobial agents and appeared to replace CG2 isolates as the dominant clones from 2003 to 2007. Apart from several canine and feline isolates that demonstrated clonality, PFGE profiles tended to be divergent across species. Whilst MDR E. coli isolates from extraintestinal infections in different animal species are diverse, some dominant CGs may persist over several years.

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