scholarly journals Tracking of antibiotic resistance transfer and rapid plasmid evolution in a hospital setting by Nanopore sequencing

2019 ◽  
Author(s):  
Silke Peter ◽  
Mattia Bosio ◽  
Caspar Gross ◽  
Daniela Bezdan ◽  
Javier Gutierrez ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Multidrug Resistance ◽  
Rapid Evolution ◽  
Hospital Setting ◽  
Gene Cassette ◽  
Multidrug Resistant ◽  
Nanopore Sequencing ◽  
Multidrug Resistance Gene ◽  
Plasmid Evolution ◽  
Resistance Gene Cassette

AbstractBackgroundInfection of patients with multidrug-resistant (MDR) bacteria often leave very limited or no treatment options. The transfer of antimicrobial resistance genes (ARG) carrying plasmids between bacterial species by horizontal gene transfer represents an important mode of expansion of ARGs. Here, we evaluated the application of Nanopore sequencing technology in a hospital setting for monitoring the transfer and rapid evolution of antibiotic resistance plasmids within and across multiple species.ResultsIn 2009 we experienced an outbreak with an extensively multidrug resistant P. aeruginosa harboring the carbapenemase enzyme blaIMP-8, and in 2012 the first Citrobacter freundii and Citrobacter werkmanii harboring the same enzyme were detected. Using Nanopore and Illumina sequencing we conducted a comparative analysis of all blaIMP-8 bacteria isolated in our hospital over a 6-year period (n = 54). We developed the computational platforms pathoLogic and plasmIDent for Nanopore-based characterization of clinical isolates and monitoring of ARG transfer, comprising de-novo assembly of genomes and plasmids, polishing, QC, plasmid circularization, ARG annotation, comparative genome analysis of multiple isolates and visualization of results. Using plasmIDent we identified a 40 kb plasmid carrying blaIMP-8 in P. aeruginosa and C. freundii, verifying that plasmid transfer had occurred. Within C. freundii the plasmid underwent further evolution and plasmid fusion, resulting in a 164 kb mega-plasmid, which was transferred to C. werkmanii. Moreover, multiple rearrangements of the multidrug resistance gene cassette were detected in P. aeruginosa, including deletions and translocations of complete ARGs.ConclusionPlasmid transfer, plasmid fusion and rearrangement of the multidrug resistance gene cassette mediated the rapid evolution of opportunistic pathogens in our hospital. We demonstrated the feasibility of tracking plasmid evolution dynamics and ARG transfer in clinical settings in a timely manner. The approach will allow for successful countermeasures to contain not only clonal, but also plasmid mediated outbreaks.

Surveillance and Antimicrobial Resistance of Salmonella Strains Isolated from Slaughtered Pigs in Spain

2007 ◽  
Vol 70 (6) ◽  
pp. 1502-1506 ◽  
Author(s):  
RAFAEL JESÚS ASTORGA MÁRQUEZ ◽  
AURORA ECHEITA SALABERRIA ◽  
ALFONSO MALDONADO GARCÍA ◽  
SILVIA VALDEZATE JIMENEZ ◽  
ALFONSO CARBONERO MARTINEZ ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Multidrug Resistance ◽  
Antimicrobial Resistance ◽  
Antimicrobial Agents ◽  
Phage Type ◽  
Multidrug Resistant ◽  
Gold Coast ◽  
Resistance Patterns ◽  
Salmonella Serotypes ◽  
Slaughtered Pigs

The prevalence of and the antibiotic resistance shown by Salmonella isolated from pigs in Andalusia (southern Spain) is reported. Salmonella enterica was recovered from 40 (33%) of 121 sampled herds, and a total of 65 isolates were serotyped. The most common Salmonella serotypes were Typhimurium and Rissen (30.7% each); others included Derby (9.2%), Brandenburg (9.2%), Newport (7.7%), Bredeney (4.6%), Anatum (3.0%), Hadar (1.5%), and Goldcoast (1.5%). One strain (1.5%) belonging to the monophasic variant of the Typhimurium serotype (Salmonella 4,5,12:i:−) was also detected. Definitive phage type (DT) 104b was the most common Typhimurium phage type isolated. These Salmonella strains were resistant to various antimicrobial agents, including tetracycline (84.6%), streptomycin (69.2%), neomycin (63.0%), sulfonamides (61.5%), ampicillin (53.8%), and amoxicillin (53.8%). All isolates were fully susceptible to ceftriaxone, ciprofloxacin, and colistin. Thirty-nine strains (64%) resistant to four or more antimicrobial agents were defined as multidrug resistant. Multidrug resistance profiles were observed in Salmonella serotypes Typhimurium, Rissen, Brandenburg, Bredeney, a monophasic variant, Gold-coast, Hadar, and Anatum, with serotypes Typhimurium and Brandenburg showing the most complicated resistance patterns (resistant to ≥11 drugs).

scholarly journals Complete Nucleotide Sequence of Two Multidrug-Resistant IncR Plasmids from Klebsiella pneumoniae

2014 ◽  
Vol 58 (7) ◽  
pp. 4207-4210 ◽  
Author(s):  
Fabrice Compain ◽  
Lionel Frangeul ◽  
Laurence Drieux ◽  
Charlotte Verdet ◽  
Sylvain Brisse ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Multidrug Resistance ◽  
Nucleotide Sequence ◽  
Resistance Genes ◽  
Complete Nucleotide Sequence ◽  
Clinical Importance ◽  
Antibiotic Resistance Genes ◽  
Multidrug Resistant ◽  
Resistance Determinants ◽  
Gene Carriers

ABSTRACTWe report here the complete nucleotide sequence of two IncR replicons encoding multidrug resistance determinants, including β-lactam (blaDHA-1,blaSHV-12), aminoglycoside (aphA1,strA,strB), and fluoroquinolone (qnrB4,aac6′-1b-cr) resistance genes. The plasmids have backbones that are similar to each other, including the replication and stability systems, and contain a wide variety of transposable elements carrying known antibiotic resistance genes. This study confirms the increasing clinical importance of IncR replicons as resistance gene carriers.

scholarly journals Multidrug Resistant Enteric Bacterial Pathogens in a Psychiatric Hospital in Ghana: Implications for Control of Nosocomial Infections

2017 ◽  
Vol 2017 ◽  
pp. 1-6 ◽  
Author(s):  
Kwabena O. Duedu ◽  
George Offei ◽  
Francis S. Codjoe ◽  
Eric S. Donkor
Keyword(s):  
Antibiotic Resistance ◽  
Multidrug Resistance ◽  
Psychiatric Hospital ◽  
Nosocomial Infections ◽  
Public Health Problem ◽  
Enteric Bacteria ◽  
Multidrug Resistant ◽  
Diffusion Method ◽  
Important Public Health Problem ◽  
Microbiological Methods

Enteric bacteria are commonly implicated in hospital-acquired or nosocomial infections. In Ghana, these infections constitute an important public health problem but little is known about their contribution to antibiotic resistance. The aim of the study was to determine the extent and pattern of antibiotic resistance of enteric bacteria isolated from patients and environmental sources at the Accra Psychiatric Hospital. A total of 265 samples were collected from the study site including 142 stool and 82 urine samples from patients, 7 swab samples of door handle, and 3 samples of drinking water. Enteric bacteria were isolated using standard microbiological methods. Antibiograms of the isolates were determined using the disc diffusion method. Overall, 232 enteric bacteria were isolated. Escherichia coli was the most common (38.3%), followed by Proteus (19.8%), Klebsiella (17.7%), Citrobacter (14.7%), Morganella (8.2%), and Pseudomonas (1.3%). All isolates were resistant to ampicillin but sensitive to cefotaxime. The resistance ranged from 15.5% to 84.5%. Multidrug resistance was most prevalent (100%) among isolates of Proteus and Morganella and least prevalent among isolates of Pseudomonas (33.3%). Multidrug resistance among enteric bacteria at the study hospital is high and hence there is a need for screening before therapy to ensure prudent use of antibiotics.

scholarly journals Functional and genetic characterization of an incF-type multidrug resistance plasmid isolated from fresh spinach

2018 ◽  
Author(s):  
Adeyinka O. Ajayi ◽  
Benjamin J. Perry ◽  
Christopher K. Yost
Keyword(s):  
Antibiotic Resistance ◽  
Multidrug Resistance ◽  
Food Safety ◽  
Sequence Analysis ◽  
Dna Sequence ◽  
Resistance Genes ◽  
Plasmid Dna ◽  
Antibiotic Resistance Genes ◽  
Multidrug Resistant ◽  
Leafy Greens

AbstractThe presence of antibiotic-resistant bacteria and clinically-relevant antibiotic resistance genes within raw foods is an on-going food safety concern. It is particularly important to be aware of the microbial quality of fresh produce because foods such as leafy greens including lettuce and spinach are minimally processed and often consumed raw therefore they often lack a microbial inactivation step. This study characterizes the genetic and functional aspects of a mobile, multidrug resistance plasmid, pLGP4, isolated from fresh spinach bought from a farmers’ market. pLGP4 was isolated using a bacterial conjugation approach. The functional characteristics of the plasmid were determined using multidrug resistance profiling and plasmid stability assays. pLGP4 was resistant to six of the eight antibiotics tested and included ciprofloxacin and meropenem. The plasmid was stably maintained within host strains in the absence of an antibiotic selection. The plasmid DNA was sequenced using an Illumina MiSeq high throughput sequencing approach and assembled into contigs using SPAdes. PCR mapping and Sanger DNA sequencing of PCR amplicons was used to complete the plasmid DNA sequence. Comparative sequence analysis determined that the plasmid was similar to plasmids that have been frequently associated with multidrug resistant clinical isolates of Klebsiella spp. DNA sequence analysis showed pLGP4 harboured qnrB1 and several other antibiotic resistance genes including three β-lactamases: blaTEM-1, blaCTX-M-15 and blaOXA-1. The detection of a multidrug-resistant, clinically-relevant plasmid on fresh spinach emphasizes the importance for vegetable producers to implement evidence-based food safety approaches into their production practises to ensure the food safety of leafy greens.

scholarly journals Epidemiology of Staphylococcus pseudintermedius in cats in Poland

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
K. Bierowiec ◽  
M. Miszczak ◽  
A. Korzeniowska-Kowal ◽  
A. Wzorek ◽  
D. Płókarz ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Multidrug Resistance ◽  
Biofilm Formation ◽  
Clinical Signs ◽  
Multidrug Resistant ◽  
Health Condition ◽  
Staphylococcus Pseudintermedius ◽  
First Time ◽  
Sequence Types ◽  
Antibiotic Resistance Properties

AbstractStaphylococcus pseudintermedius is a well-known coagulase-positive staphylococcus that is mainly associated with the asymptomatic colonization of the skin of pets and mucous membranes. Little is still known about the occurrence of S. pseudintermedius in cats. The current study aimed to characterize the isolates of S. pseudintermedius from sick and healthy cats. This was achieved by examining their antibiotic resistance properties, biofilm formation, and genotype differences. Six hundred and seventy-six cats were swabbed (595 healthy and 81 sick cats). Thirty-five distinct S. pseudintermedius isolates from 27 cats were isolated. The prevalence of S. pseudintermedius in healthy and sick cats was 2.49% and 7.61%, respectively. In comparison, MRSP (methicillin-resistant Staphylococcus pseudintermedius) prevalence was 0.12% and 2.98%, respectively. Cats were more frequently colonized with S. pseudintermedius when kept with dogs, regardless of their health condition, with this result being statistically significant. Multidrug resistance was detected in 50%, and 38.46% of S. pseudintermedius isolates from healthy and sick cats, respectively. In contrast, genetic multidrug resistance was detected in 59% and 46.15% cases, respectively. Seven from eight isolated MRSPs were multidrug-resistant. Multi-locus sequence typing (MLST) assigned isolates to 19 types, of which 16 types submitted for the first time to the PubMLST database. The most frequently detected STs (sequence types) were 551 and 71. ST71 and ST551 were mainly isolated from cats with clinical signs of infection. All were MRSPs, regardless of cats’ health. These isolates were characterized with the most frequent antibiotic resistance at the phenotypic and genotypic level.

Antibiotic Resistance Conferred by a Conjugative Plasmid and a Class I Integron in Vibrio cholerae O1 El Tor Strains Isolated in Albania and Italy

1999 ◽  
Vol 43 (3) ◽  
pp. 693-696 ◽  
Author(s):  
Vincenzo Falbo ◽  
Alessandra Carattoli ◽  
Fabio Tosini ◽  
Cristina Pezzella ◽  
Anna Maria Dionisi ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Vibrio Cholerae ◽  
Molecular Basis ◽  
Gene Cassette ◽  
Multidrug Resistant ◽  
Conjugative Plasmid ◽  
Class 1 Integron ◽  
Class 1 ◽  
Vibrio Cholerae O1 ◽  
El Tor

ABSTRACT Multidrug-resistant Vibrio cholerae O1 El Tor strains isolated during the 1994 outbreak of cholera in Albania and Italy were characterized for the molecular basis of antibiotic resistance. All strains were found to be resistant to tetracycline, streptomycin, spectinomycin, trimethoprim, sulfathiazole, and the vibriostatic compound O/129 (2,4-diamino-6,7-diisopropylteridine). Resistance genes were self-transferable by a conjugative plasmid of about 60 MDa, with the exception of spectinomycin resistance, which was conferred by theaadA1 gene cassette located in the bacterial chromosome within a class 1 integron. The resistance to trimethoprim and O/129 was conferred by the dfrA1 gene, which was present on the plasmid. Although the dfrA1 gene is known to be borne on an integron cassette, class 1, 2, or 3 intI genes were not detected as part of the plasmid DNA from the strains studied.

Salmonella in Raw Chicken Meat from the Romanian Seaside: Frequency of Isolation and Antibiotic Resistance

2015 ◽  
Vol 78 (5) ◽  
pp. 1003-1006 ◽  
Author(s):  
EMIL TÎRZIU ◽  
ROMEU LAZĂR ◽  
CLAUDIA SALA ◽  
ILEANA NICHITA ◽  
ADRIANA MORAR ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Multidrug Resistance ◽  
Antimicrobial Susceptibility ◽  
Multidrug Resistant ◽  
Chicken Meat ◽  
Contamination Rate ◽  
Fresh Meat ◽  
Retail Markets ◽  
Resistance Patterns ◽  
Meat Samples

The purpose of this study was to investigate the occurrence of Salmonella in raw chicken meat samples collected at the Romanian seaside and to evaluate the antimicrobial susceptibility of the isolates. In 2012, 317 chicken meat samples from slaughterhouses (n = 289) and retail markets (n = 28) were evaluated. Overall, 13.2% (42) of the samples contained Salmonella; 12.8% (37) and 17.8% (5) from the chicken carcasses at slaughterhouses and the fresh meat from retail markets, respectively. Eight serotypes of Salmonella enterica subsp. enterica were identified: Infantis (18 isolates), Bredeney (7), Virchow (6), Djugu (4), Grampian (4), Brandenburg (1), Derby (1), and Ruzizi (1). The isolates were resistant to tetracycline (66.6% of isolates), nalidixic acid (64.3%), sulfamethoxazole (64.3%), ciprofloxacin (61.9%), streptomycin (59.5%), trimethoprim (33.3%), ampicillin (9.5%), chloramphenicol (7.1%), and gentamicin (2.4%). No resistance was found to cefotaxime and ceftazidime. Thirty (71.4%) of the 42 tested isolates had multidrug resistance patterns to at least two antimicrobials. This survey highlighted a multidrug-resistant Salmonella contamination rate in raw chicken meat in this area of Romania, which can seriously threaten human health.

scholarly journals Antibiotic Resistance Genes and Salmonella Genomic Island 1 in Salmonella enterica Serovar Typhimurium Isolated in Italy

2002 ◽  
Vol 46 (9) ◽  
pp. 2821-2828 ◽  
Author(s):  
Alessandra Carattoli ◽  
Emma Filetici ◽  
Laura Villa ◽  
Anna Maria Dionisi ◽  
Antonia Ricci ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Salmonella Enterica ◽  
Genomic Island ◽  
Antibiotic Resistance Genes ◽  
Tetracycline Resistance ◽  
Gene Cassette ◽  
Multidrug Resistant ◽  
Phage Types ◽  
Serovar Typhimurium

ABSTRACT Fifty-four epidemiologically unrelated multidrug-resistant Salmonella enterica serovar Typhimurium isolates, collected between 1992 and 2000 in Italy, were analyzed for the presence of integrons. Strains were also tested for Salmonella genomic island 1 (SGI1), carrying antibiotic resistance genes in DT104 strains. A complete SGI1 was found in the majority of the DT104 strains. Two DT104 strains, showing resistance to streptomycin-spectinomycin and sulfonamides, carried a partially deleted SGI1 lacking the flost , tetR, and tetA genes, conferring chloramphenicol-florfenicol and tetracycline resistance, and the integron harboring the pse-1 gene cassette, conferring ampicillin resistance. The presence of SGI1 was also observed in serovar Typhimurium strains belonging to other phage types, suggesting either the potential mobility of this genomic island or changes in the phage-related phenotype of DT104 strains.

scholarly journals AcGI1, a novel genomic island carrying antibiotic resistance integron In687 in multidrug resistant Achromobacter xylosoxidans in a teaching hospital in Thailand

2020 ◽  
Vol 367 (14) ◽  
Author(s):  
Pisut Pongchaikul ◽  
Pitak Santanirand ◽  
Svetlana Antonyuk ◽  
Craig Winstanley ◽  
Alistair C Darby
Keyword(s):  
Antibiotic Resistance ◽  
Genetic Basis ◽  
Genomic Island ◽  
Hospital Setting ◽  
Carbapenem Resistance ◽  
Multidrug Resistant ◽  
Achromobacter Xylosoxidans ◽  
Whole Genome ◽  
Identical Nucleotide ◽  
Identical Nucleotide Sequence

ABSTRACT This study investigated the genetic basis of multidrug resistance in two strains of Achromobacter xylosoxidans isolated from patients attending a hospital in Thailand in 2012. These isolates were highly resistant to cephalosporins, aminoglycosides, fluoroquinolones, co-trimoxazole and carbapenems. Whole genome sequencing revealed that the two isolates were not clonally related and identified a carbapenem resistance gene-habouring integron (In687), residing in a novel genomic island, AcGI1. This In687 shares 100% identical nucleotide sequence with ones found in Acinetobacter baumannii Aci 16, isolated from the same hospital in 2007. We report the first analysis of multidrug-resistant A. xylosoxidans isolated in Thailand, and the first example of this island in A. xylosoxidans. Our data support the idea that resistance has spread in Thailand via horizontal gene transfer between species and suggest the possibility of A. xylosoxidans may serve as a reservoir of antibiotic resistance, especially in hospital setting.

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