scholarly journals Mobilome-driven segregation of the resistome in biological wastewater treatment

2021 ◽  
Author(s):  
Laura de Nies ◽  
Susheel Bhanu Busi ◽  
Benoit Josef Kunath ◽  
Patrick May ◽  
Paul Wilmes
Keyword(s):  
Wastewater Treatment ◽  
Antimicrobial Resistance ◽  
Resistance Genes ◽  
Essential Elements ◽  
Biological Wastewater Treatment ◽  
Longitudinal Assessment ◽  
Aminoglycoside Resistance ◽  
Antimicrobial Resistance Genes ◽  
Sulfonamide Resistance ◽  
Sulfonamide Resistance Genes

Biological wastewater treatment plants (BWWTP) are considered to be hotspots of evolution and subsequent spread of antimicrobial resistance (AMR). Mobile genetic elements (MGEs) promote the mobilization and dissemination of antimicrobial resistance genes (ARGs) and are thereby critical mediators of AMR within the BWWTP microbial community. At present, it is unclear whether specific AMR categories are differentially disseminated via bacteriophages (phages) or plasmids. To understand the segregation of AMR in relation to MGEs, we analyzed meta-omic (metagenomic, metatranscriptomic and metaproteomic) data systematically collected over 1.5 years from a BWWTP. Our results showed a core group of fifteen AMR categories which were found across all timepoints. Some of these AMR categories were disseminated exclusively (bacitracin) or primarily (aminoglycoside, MLS, sulfonamide) via plasmids or phages (fosfomycin and peptide), whereas others were disseminated equally by both MGEs. Subsequent expression- and protein-level analyses further demonstrated that aminoglycoside, bacitracin and sulfonamide resistance genes were expressed more by plasmids, in contrast to fosfomycin and peptide AMR expression by phages, thereby validating our genomic findings. Longitudinal assessment further underlined these findings whereby the log2-fold changes of aminoglycoside, bacitracin and sulfonamide resistance genes were increased in plasmids, while fosfomycin and peptide resistance showed similar trends in phages. In the analyzed communities, the dominant taxon Candidatus Microthrix parvicella was a major contributor to several AMR categories whereby its plasmids primarily mediated aminoglycoside resistance. Importantly, we also found AMR associated with ESKAPEE pathogens within the BWWTP, for which MGEs also contributed differentially to the dissemination of ARGs. Collectively our findings pave the way towards understanding the segmentation of AMR within MGEs, thereby shedding new light on resistome populations and their mediators, essential elements that are of immediate relevance to human health.

scholarly journals Quantifying and predicting antimicrobials and antimicrobial resistance genes in waterbodies through a holistic approach: a study in Minnesota, United States

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Irene Bueno ◽  
Amanda Beaudoin ◽  
William A. Arnold ◽  
Taegyu Kim ◽  
Lara E. Frankson ◽  
...  
Keyword(s):  
Wastewater Treatment ◽  
Antimicrobial Resistance ◽  
Resistance Genes ◽  
Wastewater Treatment Plants ◽  
Spatial Scales ◽  
Holistic Approach ◽  
Antimicrobial Resistance Genes ◽  
Study Power ◽  
High Uncertainty

AbstractThe environment plays a key role in the spread and persistence of antimicrobial resistance (AMR). Antimicrobials and antimicrobial resistance genes (ARG) are released into the environment from sources such as wastewater treatment plants, and animal farms. This study describes an approach guided by spatial mapping to quantify and predict antimicrobials and ARG in Minnesota’s waterbodies in water and sediment at two spatial scales: macro, throughout the state, and micro, in specific waterbodies. At the macroscale, the highest concentrations across all antimicrobial classes were found near populated areas. Kernel interpolation provided an approximation of antimicrobial concentrations and ARG abundance at unsampled locations. However, there was high uncertainty in these predictions, due in part to low study power and large distances between sites. At the microscale, wastewater treatment plants had an effect on ARG abundance (sul1 and sul2 in water; blaSHV, intl1, mexB, and sul2 in sediment), but not on antimicrobial concentrations. Results from sediment reflected a long-term history, while water reflected a more transient record of antimicrobials and ARG. This study highlights the value of using spatial analyses, different spatial scales, and sampling matrices, to design an environmental monitoring approach to advance our understanding of AMR persistence and dissemination.

Role of wastewater treatment plants on environmental abundance of Antimicrobial Resistance Genes in Chilean rivers

2020 ◽  
Vol 223 (1) ◽  
pp. 56-64 ◽  
Author(s):  
Irene Bueno ◽  
Claudio Verdugo ◽  
Omar Jimenez-Lopez ◽  
Pedro Pablo Alvarez ◽  
Gerardo Gonzalez-Rocha ◽  
...  
Keyword(s):  
Wastewater Treatment ◽  
Antimicrobial Resistance ◽  
Resistance Genes ◽  
Wastewater Treatment Plants ◽  
Antimicrobial Resistance Genes

scholarly journals Associations between Antimicrobial Resistance Genes in Fecal Generic Escherichia coli Isolates from Cow-Calf Herds in Western Canada

2008 ◽  
Vol 74 (12) ◽  
pp. 3658-3666 ◽  
Author(s):  
Sheryl P. Gow ◽  
Cheryl L. Waldner ◽  
Josee Harel ◽  
Patrick Boerlin
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Resistance ◽  
Resistance Genes ◽  
Gene Selection ◽  
Western Canada ◽  
Genetic Determinants ◽  
Aminoglycoside Resistance ◽  
Antimicrobial Resistance Genes ◽  
Cow Calf ◽  
Aminoglycoside Resistance Genes

ABSTRACT The objective of this study was to examine associations among the genetic determinants of antimicrobial resistance (AMR) in 207 fecal generic Escherichia coli isolates obtained from 77 cow-calf herds in western Canada. Twenty-three resistance genes corresponding to six different antimicrobial families were assessed using DNA hybridization and PCR. The most common resistance genes in the study sample (207 isolates) were sul2 (48.3%), tet(B) (45.4%), and ant(3″)-Ia (aadA1) (19.3%). Several statistically significant associations between the examined resistance genes were detected. The strongest associations observed were those between genes for resistance to chloramphenicol (catI) and trimethoprim (dhfrI) (odds ratio [OR] = 214; P = 0.0001), sulfonamide (sul1) and chloramphenicol (catI) (OR = 96.9; P = 0.0001), streptomycin [ant(3″)-Ia (aadA1)] and trimethoprim (dhfrI) (OR = 96.2; P = 0.0001), sulfonamide (sul1) and streptomycin [ant(3″)-Ia (aadA1)] (OR = 79.3; P = 0.0001), and tetracycline [tet(B)] and sulfonamides (sul2) (OR = 25.7; P = 0.0001). At least one of the resistance genes corresponding to each nonaminoglycoside family of antimicrobials examined in this study was associated with the two aminoglycoside resistance genes ant(3″)-Ia (aadA1) and aph(3′)-Ia. The multiple, strong associations between genes and the diverse nature of the associations described in this study demonstrate the complexity of resistance gene selection in cow-calf herds and should be considered in the planning of AMR control practices for cow-calf operations.

scholarly journals Extensive antimicrobial resistance mobilization via multicopy plasmid encapsidation mediated by temperate phages

2020 ◽  
Vol 75 (11) ◽  
pp. 3173-3180
Author(s):  
Lorena Rodríguez-Rubio ◽  
Carlos Serna ◽  
Manuel Ares-Arroyo ◽  
Bosco R Matamoros ◽  
Jose F Delgado-Blas ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Resistance Genes ◽  
High Efficiency ◽  
Marker Gene ◽  
Multicopy Plasmid ◽  
Aminoglycoside Resistance ◽  
Plasmid Copy ◽  
Antimicrobial Resistance Genes ◽  
Complete Sequences ◽  
High Level

Abstract Objectives To investigate the relevance of multicopy plasmids in antimicrobial resistance and assess their mobilization mediated by phage particles Methods Several databases with complete sequences of plasmids and annotated genes were analysed. The 16S methyltransferase gene armA conferring high-level aminoglycoside resistance was used as a marker in eight different plasmids, from different incompatibility groups, and with differing sizes and plasmid copy numbers. All plasmids were transformed into Escherichia coli bearing one of four different lysogenic phages. Upon induction, encapsidation of armA in phage particles was evaluated using qRT–PCR and Southern blotting. Results Multicopy plasmids carry a vast set of emerging clinically important antimicrobial resistance genes. However, 60% of these plasmids do not bear mobility (MOB) genes. When carried on these multicopy plasmids, mobilization of a marker gene armA into phage capsids was up to 10000 times more frequent than when it was encoded by a large plasmid with a low copy number. Conclusions Multicopy plasmids and phages, two major mobile genetic elements (MGE) in bacteria, represent a novel high-efficiency transmission route of antimicrobial resistance genes that deserves further investigation.

scholarly journals Draft Genome Sequence of an Escherichia coli Strain Harboring blaCTX-M-115, blaCMY-2, Aminoglycoside, Tetracycline, and Sulfonamide Resistance Genes, Isolated from a Costa Rican Wastewater Treatment Plant

2020 ◽  
Vol 9 (1) ◽  
Author(s):  
Kenia Barrantes ◽  
Luz María Chacón ◽  
Eric Morales ◽  
Lisbeth Ramírez-Carvajal
Keyword(s):  
Escherichia Coli ◽  
Wastewater Treatment ◽  
Genome Sequence ◽  
Resistance Genes ◽  
Wastewater Treatment Plant ◽  
Draft Genome ◽  
Treatment Plant ◽  
Draft Genome Sequence ◽  
Sulfonamide Resistance ◽  
Sulfonamide Resistance Genes

We report the draft genome sequence of the multidrug-resistant Escherichia coli strain PTA A1517-5, isolated from a wastewater treatment plant in Costa Rica. The genome consists of 4,927,375 bp with a GC content of 50.57% and a total of 4,853 genes. This strain harbors bla CTX-M-115, bla CMY-2, aminoglycoside, tetracycline, and sulfonamide resistance genes.

scholarly journals Culture-Independent Genotyping, Virulence and Antimicrobial Resistance Gene Identification of Staphylococcus aureus from Orthopaedic Implant-Associated Infections

2021 ◽  
Vol 9 (4) ◽  
pp. 707
Author(s):  
J. Christopher Noone ◽  
Fabienne Antunes Ferreira ◽  
Hege Vangstein Aamot
Keyword(s):  
Staphylococcus Aureus ◽  
Antimicrobial Resistance ◽  
Resistance Genes ◽  
Virulence Gene ◽  
Orthopaedic Implant ◽  
Metagenomic Sequencing ◽  
Gene Detection ◽  
Shotgun Metagenomic Sequencing ◽  
Antimicrobial Resistance Genes ◽  
Culture Independent

Our culture-independent nanopore shotgun metagenomic sequencing protocol on biopsies has the potential for same-day diagnostics of orthopaedic implant-associated infections (OIAI). As OIAI are frequently caused by Staphylococcus aureus, we included S. aureus genotyping and virulence gene detection to exploit the protocol to its fullest. The aim was to evaluate S. aureus genotyping, virulence and antimicrobial resistance genes detection using the shotgun metagenomic sequencing protocol. This proof of concept study included six patients with S. aureus-associated OIAI at Akershus University Hospital, Norway. Five tissue biopsies from each patient were divided in two: (1) conventional microbiological diagnostics and genotyping, and whole genome sequencing (WGS) of S. aureus isolates; (2) shotgun metagenomic sequencing of DNA from the biopsies. Consensus sequences were analysed using spaTyper, MLST, VirulenceFinder, and ResFinder from the Center for Genomic Epidemiology (CGE). MLST was also compared using krocus. All spa-types, one CGE and four krocus MLST results matched Sanger sequencing results. Virulence gene detection matched between WGS and shotgun metagenomic sequencing. ResFinder results corresponded to resistance phenotype. S. aureus spa-typing, and identification of virulence and antimicrobial resistance genes are possible using our shotgun metagenomics protocol. MLST requires further optimization. The protocol has potential application to other species and infection types.

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