scholarly journals Complete assembly ofEscherichia coliST131 genomes using long reads demonstrates antibiotic resistance gene variation within diverse plasmid and chromosomal contexts

2019 ◽  
Author(s):  
Arun Decano ◽  
Catherine Ludden ◽  
Theresa Feltwell ◽  
Kim Judge ◽  
Julian Parkhill ◽  
...  

AbstractThe incidence of infections caused by extraintestinalEscherichia coli(ExPEC) is rising globally, which is a major public health concern. ExPEC strains that are resistant to antimicrobials have been associated with excess mortality, prolonged hospital stays and higher healthcare costs.E. coliST131 is a major ExPEC clonal group worldwide with variable plasmid composition, and has an array of genes enabling antimicrobial resistance (AMR). ST131 isolates frequently encode the AMR genesblaCTX-M-14/15/27, which are often rearranged, amplified and translocated by mobile genetic elements (MGEs). Short DNA reads do not fully resolve the architecture of repetitive elements on plasmids to allow MGE structures encodingblaCTX-Mgenes to be fully determined. Here, we performed long read sequencing to decipher the genome structures of sixE. coliST131 isolated from six patients. Most long read assemblies generated entire chromosomes and plasmids as single contigs, contrasting with more fragmented assemblies created with short reads alone. The long read assemblies highlighted diverse accessory genomes withblaCTX-M-15,blaCTX-M-14andblaCTX-M-27genes identified in three, one and one isolates, respectively. One sample had noblaCTX-Mgene. Two samples had chromosomalblaCTX-M-14andblaCTX-M-15genes, and the latter was at three distinct locations, likely transposed by the adjacent MGEs: ISEcp1, IS903Band Tn2. This study showed that AMR genes exist in multiple different chromosomal and plasmid contexts even between closely-related isolates within a clonal group such asE. coliST131.ImportanceDrug-resistant bacteria are a major cause of illness worldwide and a specific subtype calledEscherichia coliST131 cause a significant amount of these infections. ST131 become resistant to treatment by modifying their DNA and by transferring genes among one another via large packages of genes called plasmids, like a game of pass-the-parcel. Tackling infections more effectively requires a better understanding of what plasmids are being exchanged and their exact contents. To achieve this, we applied new high-resolution DNA sequencing technology to six ST131 samples from infected patients and compared the output to an existing approach. A combination of methods shows that drug-resistance genes on plasmids are highly mobile because they can jump into ST131’s chromosomes. We found that the plasmids are very elastic and undergo extensive rearrangements even in closely related samples. This application of DNA sequencing technologies illustrates at a new level the highly dynamic nature of ST131 genomes.

mSphere ◽  
2019 ◽  
Vol 4 (3) ◽  
Author(s):  
Arun Gonzales Decano ◽  
Catherine Ludden ◽  
Theresa Feltwell ◽  
Kim Judge ◽  
Julian Parkhill ◽  
...  

ABSTRACTThe incidence of infections caused by extraintestinalEscherichia coli(ExPEC) is rising globally, which is a major public health concern. ExPEC strains that are resistant to antimicrobials have been associated with excess mortality, prolonged hospital stays, and higher health care costs.E. colisequence type 131 (ST131) is a major ExPEC clonal group worldwide, with variable plasmid composition, and has an array of genes enabling antimicrobial resistance (AMR). ST131 isolates frequently encode the AMR genesblaCTX-M-14,blaCTX-M-15, andblaCTX-M-27, which are often rearranged, amplified, and translocated by mobile genetic elements (MGEs). Short DNA reads do not fully resolve the architecture of repetitive elements on plasmids to allow MGE structures encodingblaCTX-Mgenes to be fully determined. Here, we performed long-read sequencing to decipher the genome structures of sixE. coliST131 isolates from six patients. Most long-read assemblies generated entire chromosomes and plasmids as single contigs, in contrast to more fragmented assemblies created with short reads alone. The long-read assemblies highlighted diverse accessory genomes withblaCTX-M-15,blaCTX-M-14, andblaCTX-M-27genes identified in three, one, and one isolates, respectively. One sample had noblaCTX-Mgene. Two samples had chromosomalblaCTX-M-14andblaCTX-M-15genes, and the latter was at three distinct locations, likely transposed by the adjacent MGEs: ISEcp1, IS903B,and Tn2. This study showed that AMR genes exist in multiple different chromosomal and plasmid contexts, even between closely related isolates within a clonal group such asE. coliST131.IMPORTANCEDrug-resistant bacteria are a major cause of illness worldwide, and a specific subtype calledEscherichia coliST131 causes a significant number of these infections. ST131 bacteria become resistant to treatments by modifying their DNA and by transferring genes among one another via large packages of genes called plasmids, like a game of pass-the-parcel. Tackling infections more effectively requires a better understanding of what plasmids are being exchanged and their exact contents. To achieve this, we applied new high-resolution DNA sequencing technology to six ST131 samples from infected patients and compared the output to that of an existing approach. A combination of methods shows that drug resistance genes on plasmids are highly mobile because they can jump into ST131’s chromosomes. We found that the plasmids are very elastic and undergo extensive rearrangements even in closely related samples. This application of DNA sequencing technologies illustrates at a new level the highly dynamic nature of ST131 genomes.


2021 ◽  
Vol 16 (1) ◽  
Author(s):  
Neha Giri ◽  
Anchal Lodhi ◽  
Devendra Singh Bisht ◽  
Suvarna Bhoj ◽  
Deepak Kumar Arya

Researchers have encountered new challenges with the discovery of multiple drug resistance in microbes. Currently, multidrug resistant bacteria are considered a major public health concern and an emerging global epidemic. Presence of Escherichia coli in water is used as a faecal pollution measure. In this study E. coli isolates were collected from 20 sample collection sites at Lake Nainital. 20 E. coli isolates, 1 from each sample collection sites, were examined for their antibiotic response patterns against a panel of widely used 15 antibiotics. The result of this study showed 100% resistance to Penicillin G followed by Erythromycin (80%). All isolates (100%) were found susceptible for Gentamycin. The susceptibilities for Chloramphenicol and Co-trimoxazaole were found next to Gentamycin as 90 and 85% respectively. Multiple antibiotic resistance (MAR) index was also determined. 0.73 MAR index was observed as highest in 1 isolate. 13 out of 20 isolates had more than 0.2 MAR indices. The result reveals the origin of E. coli isolates from an area of high antibiotics use.


2010 ◽  
Vol 76 (21) ◽  
pp. 6991-6997 ◽  
Author(s):  
Azucena Mora ◽  
Alexandra Herrera ◽  
Rosalia Mamani ◽  
Cecilia López ◽  
María Pilar Alonso ◽  
...  

ABSTRACT To discern the possible spread of the Escherichia coli O25b:H4-ST131 clonal group in poultry and the zoonotic potential of avian strains, we made a retrospective search of our strain collection and compared the findings for those strains with the findings for current strains. Thus, we have characterized a collection of 19 avian O25b:H4-ST131 E. coli strains isolated from 1995 to 2010 which, interestingly, harbored the ibeA gene. Using this virulence gene as a criterion for selection, we compared those 19 avian strains with 33 human O25b:H4-ST131 ibeA-positive E. coli strains obtained from patients with extraintestinal infections (1993 to 2009). All 52 O25b:H4-ST131 ibeA-positive E. coli strains shared the fimH, kpsMII, malX, and usp genes but showed statistically significant differences in nine virulence factors, namely, papGIII, cdtB, sat, and kpsMII K5, which were associated with human strains, and iroN, kpsMII K1, cvaC, iss, and tsh, which were associated with strains of avian origin. The XbaI macrorestriction profiles of the 52 E. coli O25b:H4-ST131 ibeA-positive strains revealed 11 clusters (clusters I to XI) of >85% similarity, with four clusters including strains of human and avian origin. Cluster VII (90.9% similarity) grouped 10 strains (7 avian and 3 human strains) that mostly produced CTX-M-9 and that also shared the same virulence profile. Finally, we compared the macrorestriction profiles of the 12 CTX-M-9-producing O25b:H4-ST131 ibeA strains (7 avian and 5 human strains) identified among the 52 strains with those of 15 human O25b:H4-ST131 CTX-M-14-, CTX-M-15-, and CTX-M-32-producing strains that proved to be negative for ibeA and showed that they clearly differed in the level of similarity from the CTX-M-9-producing strains. In conclusion, E. coli clonal group O25b:H4-ST131 ibeA has recently emerged among avian isolates with the new acquisition of the K1 capsule antigen and includes CTX-M-9-producing strains. This clonal group represents a real zoonotic risk that has crossed the barrier between human and avian hosts.


2019 ◽  
Vol 2019 ◽  
pp. 1-8 ◽  
Author(s):  
Mourouge Saadi Alwash ◽  
Hawraa Mohammed Al-Rafyai

Surface water contamination remains a major worldwide public health concern and may contribute to the dissemination of antibiotic-resistant bacteria. The Al-Hillah River in the city of Babylon Province, Iraq, diverts flows from the Euphrates River. Because of its importance in irrigation and population density, it faces several forced and unforced changes due to anthropogenic activities. To evaluate water quality, water samples were collected from three sites with different anthropogenic pressures along the Al-Hillah River. These samples were subjected to bacteriological analyses, i.e., total coliforms, Escherichia coli, and faecal enterococci. The phylogenetic groups of the E. coli isolates (n = 61) were typed by rapid PCR-based analyses. Representatives of each isolate were tested phenotypically for resistance to six classes of antibiotics and characterized according to their phylogenetic groups. The results demonstrated the highest resistance levels were to β-lactam antibiotics, followed by fosfomycin and aminoglycosides. Escherichia coli isolates belonging to phylogenetic groups A and B2 were the most common and were characterized by a higher prevalence of antibiotic resistance. This study is important for understanding the current conditions of the Al-Hillah River, as the data reveal a high prevalence of multiresistance among E. coli isolates circulating at the three sampling sites.


Author(s):  
Asinamai Athliamai Bitrus ◽  
Peter Anjili Mshelia ◽  
Iliya Dauda Kwoji ◽  
Mohammed Dauda Goni ◽  
Saleh Mohammed Jajere

Antimicrobial resistance has gained global notoriety due to its public health concern, the emergence of multiple drug-resistant bacteria, and lack of new antimicrobials. Extended-spectrum beta-lactamase (ESBL)/ampicillin Class C (AmpC)- producing Escherichia coli and other zoonotic pathogens can be transmitted to humans from animals either through the food chain, direct contact or contamination of shared environments. There is a surge in the rate of resistance to medically important antibiotics such as carbapenem, ESBL, aminoglycosides, and fluoroquinolones among bacteria of zoonotic importance. Factors that may facilitate the occurrence, persistence and dissemination of ESBL/AmpC-Producing E. coli in humans and animal includes; 1). o ral administration of antimicrobials to humans primarily (by physician and health care providers) and secondarily to animals, 2). importation of parent stock and day-old chickens, 3). farm management practice and lack of water acidification in poultry, 4). contamination of feed, water and environment, 5). contamination of plants with feces of animals. Understanding these key factors will help reduce the level of resistance, thereby boosting the therapeutic effectiveness of antimicrobial agents in the treatment of animal and human infections. This review highlights the occurrence, risk factors, and public health importance of ESBL/AmpC-beta-lactamase producing E. coli isolated from livestock.


2020 ◽  
Vol 7 (2) ◽  
Author(s):  
Tim Flerlage ◽  
Jessica N Brazelton de Cardenas ◽  
Cherilyn D Garner ◽  
Nur A Hasan ◽  
Hiren Karathia ◽  
...  

Abstract Background Genes conferring carbapenem resistance have disseminated worldwide among Gram-negative bacteria. Here we present longitudinal changes in clinically obtained Escherichia coli isolates from 1 immunocompromised pediatric patient. This report demonstrates potential for antibiotic resistance genes and plasmids to emerge over time in clinical isolates from patients receiving intensive anticancer chemotherapy and broad-spectrum antibiotics. Methods Thirty-three isolates obtained over 7 months from 1 patient were included. Clinical data were abstracted from the medical record. For each isolate, studies included phenotypic antibacterial resistance patterns, sequence typing, bacterial isolate sequencing, plasmid identification, and antibiotic resistance gene identification. Results Sites of isolation included blood, wound culture, and culture for surveillance purposes from the perianal area. Isolates were of 5 sequence types (STs). All were resistant to multiple classes of antibiotics; 23 (69.6%) were phenotypically resistant to all carbapenems. The blaNDM-5 gene was identified in 22 (67%) isolates, all of ST-167 and ST-940, and appeared to coincide with the presence of the IncFII and IncX3 plasmid. Conclusions We present unique microbiologic data from 33 multidrug-resistant E. coli isolates obtained over the course of 7 months from an individual patient in the United States. Two E. coli sequence types causing invasive infection in the same patient and harboring the blaNDM-5 gene, encoded on the IncX3 plasmid and the IncFII plasmid, were identified. This study highlights the emergence of multidrug-resistant bacteria on antibiotic therapy and the necessity of adequate neutrophil number and function in the clearance of bacteremia.


2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Kayode Fashae ◽  
Ines Engelmann ◽  
Stefan Monecke ◽  
Sascha D. Braun ◽  
Ralf Ehricht

Abstract Background Antimicrobial resistance (AMR) is an increasing global health concern reducing options for therapy of infections and also for perioperative prophylaxis. Many Enterobacteriaceae cannot be treated anymore with third generation cephalosporins (3GC) due to the production of certain 3GC hydrolysing enzymes (extended spectrum beta-lactamases, ESBLs). The role of animals as carriers and vectors of multi-resistant bacteria in different geographical regions is poorly understood. Therefore, we investigated the occurrence and molecular characteristics of ESBL-producing Escherichia coli (E. coli) in wild birds and slaughtered cattle in Ibadan, Nigeria. Cattle faecal samples (n = 250) and wild bird pooled faecal samples (cattle egrets, Bubulcus ibis, n = 28; white-faced whistling duck, Dendrocygna viduata, n = 24) were collected and cultured on cefotaxime-eosin methylene blue agar. Antimicrobial susceptibility was determined by agar diffusion assays and all 3GC resistant isolates were genotypically characterised for AMR genes, virulence associated genes (VAGs) and serotypes using DNA microarray-based assays. Results All 3GC resistant isolates were E. coli: cattle (n = 53), egrets (n = 87) and whistling duck (n = 4); cultured from 32/250 (12.8%), 26/28 (92.9%), 2/24(8.3%), cattle, egrets and whistling duck faecal samples, respectively. blaCTX-M gene family was prevalent; blaCTX-M15 (83.3%) predominated over blaCTX-M9 (11.8%). All were susceptible to carbapenems. The majority of isolates were resistant to at least one of the other tested antimicrobials; multidrug resistance was highest in the isolates recovered from egrets. The isolates harboured diverse repositories of other AMR genes (including strB and sul2), integrons (predominantly class 1) and VAGs. The isolates recovered from egrets harboured more AMR genes; eight were unique to these isolates including tetG, gepA, and floR. The prevalent VAGs included hemL and iss; while 14 (including sepA) were unique to certain animal isolates. E. coli serotypes O9:H9, O9:H30 and O9:H4 predominated. An identical phenotypic microarray profile was detected in three isolates from egrets and cattle, indicative of a clonal relationship amongst these isolates. Conclusion Wild birds and cattle harbour diverse ESBL-producing E. coli populations with potential of inter-species dissemination and virulence. Recommended guidelines to balance public health and habitat conservation should be implemented with continuous surveillance.


2018 ◽  
Vol 3 (2) ◽  
pp. 26
Author(s):  
Asman Sadino ◽  
Idin Sahidin ◽  
Wahyuni Wahyuni

The emergence of resistant bacteria strain has become a global health concern. It encourages the exploration of potential antibacterial agents, particularly from natural sources. The aim of this study was to investigate the antibacterial activity of ethanol extract of root, stems, leaves, and flowers of Polygonum pulchrum Blume against Staphylococcus aureus and Escherichia coli, through disc diffusion method using cup-plate method. Inhibition zone against S. aureus from roots, stems, leaves, and flowers ethanol extract were 3.5 mm, 2.5 mm, 2.25 mm, and 2.62 mm, respectively, while the inhibition zone against E. coli were 2.25 mm, 2.12 mm, 1.62 mm, and 1.75 mm, respectively. In conclusion, ethanol extract of root, stem, leaves, and flower of P. pulchrum Bl possessed weak antibacterial activity against S. aureus and E. coli.Keywords: P. pulchrum Bl, antibacterial, E. coli, S. aureus, cup-plate technique


2021 ◽  
Author(s):  
Tatsuya Nakayama ◽  
Le Thi Hien ◽  
Ngo Thanh Phong ◽  
Doan Nguyen Minh Tran ◽  
Oanh Thi Hoang Nguyen ◽  
...  

Abstract Although the spread of plasmid-mediated antibiotic-resistant bacteria is a public health concern, food contamination with plasmid-mediated antibiotic-resistant Escherichia coli has not been well investigated in Vietnam. The aim of this study was to describe the prevalence of colistin-resistant, carbapenem-resistant and endemic blaCTX−M in extended-spectrum β-lactamase (ESBL)-producing E. coli isolates. Colistin- and carbapenem-resistant ESBL-producing E. coli were isolated from chickens in Vietnam and Japan. The results showed that 52% and 93% of Vietnamese chicken was isolated with colistin-resistant and AmpC/ESBL-producing E. coli, respectively, while 52.7% of Japanese chickens were isolated with AmpC/ESBL-producing E. coli. Carbapenem-resistant E. coli has not been isolated in Vietnam or Japan. Genotyping revealed that colistin-resistant E. coli harboured mcr-1, and most of the AmpC/ESBL-related genes were blaCTX−M−55 and blaCTX−M−65 together with blaTEM in Vietnamese chickens, and blaCMY−2 in Japanese chickens. Multidrug resistance analysis showed that ESBL-producing E. coli isolates were more resistant to quinolones, streptomycin, and chloramphenicol compared with colistin-resistant E. coli isolates from Vietnam, suggesting selection in ESBL-producing E. coli for multiple antibiotic resistance genes. In conclusion, colistin-resistant E. coli was detected in about half of the chicken meat samples, the majority of which were found to harbour mcr-1. The high prevalence of ESBL-producing E. coli has remained constant across the last five years, and the predominant blaCTX−M for ESBL-producing E. coli was found to be blaCTX−M−55 or blaCTX−M−65, with the coexistence of blaTEM in Vietnam. Our results can be implemented in monitoring systems to combat the development of antimicrobial resistance.


2021 ◽  
Author(s):  
Abimbola Olumide Adekanmbi ◽  
Olabisi C. Akinlabi ◽  
Adedolapo V. Olaposi

Abstract There is a rapid rise in the incident of quinolone resistant bacteria in Nigeria. Most studies in Nigeria have focused on isolates from the clinical settings, with few focusing on isolates of environmental origin. This study aimed to investigate the antibiogram and carriage of plasmid-mediated quinolone resistance (PMQR) genes by quinolone-resistant isolates obtained from a pool of cefotaxime-resistant Escherichia coli (E. coli) recovered from sewage leaking out of some broken sanitary sewers in a University community in Nigeria. Isolation of E. coli from the sewage samples was done on CHROMagar E. coli after enrichment of the samples was done in Brain Heart Infusion amended with 6µg/mL of cefotaxime. Identification of presumptive E. coli was done using molecular methods (detection of uidA gene), while susceptibility to antibiotics was carried out using disc-diffusion method. Detection of PMQR genes (qnrA, qnrB, qnrS, aac(6')-lb-cr, qepA and oqxAB) was done using primer-specific PCR. A total of 32 non-repetitive cefotaxime-resistant E. coli were obtained from the sewage, with 21 being quinolone-resistant. The quinolone-resistant isolates showed varying level of resistance to the tested antibiotics, with imipenem being the only exception with 0% resistance. The PMQR genes: aac(6')-lb-cr, qnrA, qnrB, qnrS and qepA and oqxAB were detected in 90.5%, 61.9%, 47.6%, 38.1%, 4.8% and 0% respectively of the isolates. The findings of this study showed a high level of resistance to antibiotics and carriage of PMQR genes by quinolone-resistant E. coli obtained from the leaking sanitary sewers, suggesting a potential environmental and public health concern.


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