scholarly journals Frequency of Antimicrobial Resistance Genes in Salmonella From Brazil by in silico Whole-Genome Sequencing Analysis: An Overview of the Last Four Decades

2020 ◽  
Vol 11 ◽  
Author(s):  
Grazielle Lima Rodrigues ◽  
Pedro Panzenhagen ◽  
Rafaela Gomes Ferrari ◽  
Anamaria dos Santos ◽  
Vania Margaret Flosi Paschoalin ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Resistance Genes ◽  
In Silico ◽  
Whole Genome ◽  
Sequencing Analysis ◽  
Antimicrobial Resistance Genes

scholarly journals Whole-Genome Sequencing Analysis Accurately Predicts Antimicrobial Resistance Phenotypes in Campylobacter spp.

2015 ◽  
Vol 82 (2) ◽  
pp. 459-466 ◽  
Author(s):  
S. Zhao ◽  
G. H. Tyson ◽  
Y. Chen ◽  
C. Li ◽  
S. Mukherjee ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Resistance Genes ◽  
Antimicrobial Agents ◽  
The United States ◽  
23S Rrna ◽  
Whole Genome ◽  
Sequencing Analysis ◽  
Content Type

ABSTRACTThe objectives of this study were to identify antimicrobial resistance genotypes forCampylobacterand to evaluate the correlation between resistance phenotypes and genotypes usingin vitroantimicrobial susceptibility testing and whole-genome sequencing (WGS). A total of 114Campylobacterspecies isolates (82C. coliand 32C. jejuni) obtained from 2000 to 2013 from humans, retail meats, and cecal samples from food production animals in the United States as part of the National Antimicrobial Resistance Monitoring System were selected for study. Resistance phenotypes were determined using broth microdilution of nine antimicrobials. Genomic DNA was sequenced using the Illumina MiSeq platform, and resistance genotypes were identified using assembled WGS sequences through blastx analysis. Eighteen resistance genes, includingtet(O),blaOXA-61,catA,lnu(C),aph(2″)-Ib,aph(2″)-Ic,aph(2′)-If,aph(2″)-Ig,aph(2″)-Ih,aac(6′)-Ie-aph(2″)-Ia,aac(6′)-Ie-aph(2″)-If,aac(6′)-Im,aadE,sat4,ant(6′),aad9,aph(3′)-Ic, andaph(3′)-IIIa, and mutations in two housekeeping genes (gyrAand 23S rRNA) were identified. There was a high degree of correlation between phenotypic resistance to a given drug and the presence of one or more corresponding resistance genes. Phenotypic and genotypic correlation was 100% for tetracycline, ciprofloxacin/nalidixic acid, and erythromycin, and correlations ranged from 95.4% to 98.7% for gentamicin, azithromycin, clindamycin, and telithromycin. All isolates were susceptible to florfenicol, and no genes associated with florfenicol resistance were detected. There was a strong correlation (99.2%) between resistance genotypes and phenotypes, suggesting that WGS is a reliable indicator of resistance to the nine antimicrobial agents assayed in this study. WGS has the potential to be a powerful tool for antimicrobial resistance surveillance programs.

scholarly journals Genomic Characterization of Fluoroquinolone-Resistant Thermophilic Campylobacter Strains Isolated from Layer Chicken Feces in Gangneung, South Korea by Whole-Genome Sequencing

Genes ◽  
2021 ◽  
Vol 12 (8) ◽  
pp. 1131
Author(s):  
Noel Gahamanyi ◽  
Dae-Geun Song ◽  
Kye-Yoon Yoon ◽  
Leonard E. G. Mboera ◽  
Mecky I. Matee ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Resistance Genes ◽  
Virulence Genes ◽  
Coli Strain ◽  
Whole Genome ◽  
Thermophilic Campylobacter ◽  
Antimicrobial Resistance Genes ◽  
Campylobacter Species

Thermophilic Campylobacter species of poultry origin have been associated with up to 80% of human campylobacteriosis cases. Layer chickens have received less attention as possible reservoirs of Campylobacter species. Initially, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of two archived Campylobacter isolates (Campylobacter jejuni strain 200605 and Campylobacter coli strain 200606) from layer chickens to five antimicrobials (ciprofloxacin, nalidixic acid, erythromycin, tetracycline, and gentamicin) were determined using broth microdilution while the presence of selected antimicrobial resistance genes was performed by polymerase chain reaction (PCR) using specific primers. Whole-genome sequencing (WGS) was performed by the Illumina HiSeq X platform. The analysis involved antimicrobial resistance genes, virulome, multilocus sequence typing (MLST), and phylogeny. Both isolates were phenotypically resistant to ciprofloxacin (MIC: 32 vs. 32 µg/mL), nalidixic acid (MIC: 128 vs. 64 µg/mL), and tetracycline (MIC: 64 vs. 64 µg/mL), but sensitive to erythromycin (MIC: 1 vs. 2 µg/mL) and gentamicin (MIC: 0.25 vs. 1 µg/mL) for C. jejuni strain 200605 and C. coli strain 200606, respectively. WGS confirmed C257T mutation in the gyrA gene and the presence of cmeABC complex conferring resistance to FQs in both strains. Both strains also exhibited tet(O) genes associated with tetracycline resistance. Various virulence genes associated with motility, chemotaxis, and capsule formation were found in both isolates. However, the analysis of virulence genes showed that C. jejuni strain 200605 is more virulent than C. coli strain 200606. The MLST showed that C. jejuni strain 200605 belongs to sequence type ST-5229 while C. coli strain 200606 belongs to ST-5935, and both STs are less common. The phylogenetic analysis clustered C. jejuni strain 200605 along with other strains reported in Korea (CP028933 from chicken and CP014344 from human) while C. coli strain 200606 formed a separate cluster with C. coli (CP007181) from turkey. The WGS confirmed FQ-resistance in both strains and showed potential virulence of both strains. Further studies are recommended to understand the reasons behind the regional distribution (Korea, China, and Vietnam) of such rare STs.

scholarly journals Comparative Genomic Analysis and Phenotypic Characterization of Bronchoscope-Associated Klebsiella Aerogenes

2020 ◽  
Author(s):  
Fang Huang ◽  
Shuang Li ◽  
Xiaohui Chi ◽  
Peipei Wen ◽  
Hao Fu ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Resistance Genes ◽  
Antimicrobial Susceptibility ◽  
Susceptibility Testing ◽  
Antimicrobial Susceptibility Testing ◽  
Klebsiella Aerogenes ◽  
Whole Genome ◽  
Antimicrobial Resistance Genes

Abstract Background: Bronchoscopes has been linked to the outbreaks of nosocomial infections. We aim to investigate the phenotypic and genomic profiles of bronchoscope-associated Klebsiella aerogenes isolates, and their association with genome public available isolates from human and environment.Methods: We performed a prospective single-center study sampling echoendoscopes after clinical use and after normal decontamination procedures. Bacterial screening was conducted by culturing the sample on Mueller-Hinton agar plates. Antimicrobial susceptibility testing was performed using the broth microdilution method. Whole-genome sequencing of K. aerogenes isolates was performed using an Illumina HiSeq system and comparative genomics analysis were conducted.Results: Over the 5-month period, a total of 358 isolates and 13 isolates were recovered from samples after clinical procedures and samples after decontamination procedures, respectively. Antimicrobial susceptibility testing found 7 K. aerogenes isolates to exhibit low-level resistance to antimicrobial agents. Among 7 K. aerogenes isolates, we found 5 sequence types (STs). Whole genome sequencing and comparison analysis observed the genetic diversity in our bacterial collection, which clustered into three main clades. Furthermore, we identified a total of 43 antimicrobial resistance genes in the K. aerogenes core genomes. As expected, human isolates encoded more antimicrobial resistance genes than that environmental isolates. Conclusions: This study first described the phenotypic and genomics characteristics of bronchoscope-associated K. aerogenes. The present observations demonstrated that broadly investigation of specific pathogens using publicly available global genomes offered the opportunity to identify prevalent clones associated with various hosts, sources, and geographical locations.

scholarly journals Comparison of Antimicrobial Resistance and Pan-Genome of Clinical and Non-Clinical Enterococcus cecorum from Poultry Using Whole-Genome Sequencing

Foods ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 686
Author(s):  
Poonam Sharma ◽  
Sushim K. Gupta ◽  
John B. Barrett ◽  
Lari M. Hiott ◽  
Tiffanie A. Woodley ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Resistance Genes ◽  
Gene Families ◽  
Clinical Isolates ◽  
Whole Genome ◽  
Pan Genome ◽  
Antimicrobial Resistance Genes ◽  
Enterococcus Cecorum

Enterococcus cecorum is an emerging avian pathogen, particularly in chickens, but can be found in both diseased (clinical) and healthy (non-clinical) poultry. To better define differences between E. cecorum from the two groups, whole-genome sequencing (WGS) was used to identify and compare antimicrobial resistance genes as well as the pan-genome among the isolates. Eighteen strains selected from our previous study were subjected to WGS using Illumina MiSeq and comparatively analyzed. Assembled contigs were analyzed for resistance genes using ARG-ANNOT. Resistance to erythromycin was mediated by ermB, ermG, and mefA, in clinical isolates and ermB and mefA, in non-clinical isolates. Lincomycin resistance genes were identified as linB, lnuB, lnuC, and lnuD with lnuD found only in non-clinical E. cecorum; however, lnuB and linB were found in only one clinical isolate. For both groups of isolates, kanamycin resistance was mediated by aph3-III, while tetracycline resistance was conferred by tetM, tetO, and tetL. No mutations or known resistance genes were found for isolates resistant to either linezolid or chloramphenicol, suggesting possible new mechanisms of resistance to these drugs. A comparison of WGS results confirmed that non-clinical isolates contained more resistance genes than clinical isolates. The pan-genome of clinical and non-clinical isolates resulted in 3651 and 4950 gene families, respectively, whereas the core gene sets were comprised of 1559 and 1534 gene families in clinical and non-clinical isolates, respectively. Unique genes were found more frequently in non-clinical isolates than clinical. Phylogenetic analysis of the isolates and all the available complete and draft genomes showed no correlation between healthy and diseased poultry. Additional genomic comparison is required to elucidate genetic factors in E. cecorum that contribute to disease in poultry.

scholarly journals Comparative Genomic Analysis and Phenotypic Characterization of Bronchoscope-associated Klebsiella aerogenes

2020 ◽  
Author(s):  
Fang Huang ◽  
Shuang Li ◽  
Xiaohui Chi ◽  
Peipei Wen ◽  
Hao Fu ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Resistance Genes ◽  
Antimicrobial Susceptibility ◽  
Susceptibility Testing ◽  
Antimicrobial Susceptibility Testing ◽  
Klebsiella Aerogenes ◽  
Whole Genome ◽  
Antimicrobial Resistance Genes

Abstract Background: Bronchoscopes has been linked to the outbreaks of nosocomial infections. We aim to investigate the phenotypic and genomic profiles of bronchoscope-associated Klebsiella aerogenes isolates, and their association with genome public available isolates from human and environment.Methods: We performed a prospective single-center study sampling echoendoscopes after clinical use and after normal decontamination procedures. Bacterial screening was conducted by culturing the sample on Mueller-Hinton agar plates. Antimicrobial susceptibility testing was performed using the broth microdilution method. Whole-genome sequencing of K. aerogenes isolates was performed using an Illumina HiSeq system and comparative genomics analysis were conducted.Results: Over the 5-month period, a total of 358 isolates and 13 isolates were recovered from samples after clinical procedures and samples after decontamination procedures, respectively. Antimicrobial susceptibility testing found 7 K. aerogenes isolates to exhibit low-level resistance to antimicrobial agents. Among 7 K. aerogenes isolates, we found 5 sequence types (STs). Whole genome sequencing and comparison analysis observed the genetic diversity in our bacterial collection, which clustered into three main clades. Furthermore, we identified a total of 43 antimicrobial resistance genes in the K. aerogenes core genomes. As expected, human isolates encoded more antimicrobial resistance genes than that environmental isolates. Conclusions: This study first described the phenotypic and genomics characteristics of bronchoscope-associated K. aerogenes. The present observations demonstrated that broadly investigation of specific pathogens using publicly available global genomes offered the opportunity to identify prevalent clones associated with various hosts, sources, and geographical locations.

scholarly journals WGS based analysis of acquired antimicrobial resistance in human and non-human Acinetobacter baumannii isolates from a German perspective

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Gamal Wareth ◽  
Christian Brandt ◽  
Lisa D. Sprague ◽  
Heinrich Neubauer ◽  
Mathias W. Pletz
Keyword(s):  
Antimicrobial Resistance ◽  
Acinetobacter Baumannii ◽  
Genome Sequencing ◽  
Resistance Genes ◽  
In Silico ◽  
Acquired Resistance ◽  
Whole Genome ◽  
Chromosomal Gene ◽  
Milk Samples ◽  
Carbapenemase Gene

Abstract Background Acinetobacter baumannii ability to develop and acquire resistance makes it one of the most critical nosocomial pathogens globally. Whole-genome sequencing (WGS) was applied to identify the acquired or mutational variants of antimicrobial resistance (AMR) genes in 85 German A. baumannii strains utilizing Illumina technology. Additionally, the whole genome of 104 German isolates deposited in the NCBI database was investigated. Results In-silico analysis of WGS data revealed wide varieties of acquired AMR genes mediating resistance mostly to aminoglycosides, cephalosporins, carbapenems, sulfonamides, tetracyclines and macrolides. In the 189 analyzed genomes, the ant (3″)-IIa conferring resistance to aminoglycosides was the most frequent (55%), followed by blaADC.25 (38.6%) conferring resistance to cephalosporin, blaOXA-23 (29%) and the blaOXA-66 variant of the intrinsic blaOXA-51-likes (26.5%) conferring resistance to carbapenems, the sul2 (26%) conferring resistance to sulfonamides, the tet. B (19.5%) conferring resistance to tetracycline, and mph. E and msr. E (19%) conferring resistance to macrolides. blaTEM variants conferring resistance to cephalosporins were found in 12% of genomes. Thirteen variants of the intrinsic blaOXA-51 carbapenemase gene, blaOXA-510 and blaADC-25 genes were found in isolates obtained from dried milk samples. Conclusion The presence of strains harboring acquired AMR genes in dried milk raises safety concerns and highlights the need for changes in producing dried milk. Acquired resistance genes and chromosomal gene mutation are successful routes for disseminating AMR determinants among A. baumannii. Identification of chromosomal and plasmid-encoded AMR in the genome of A. baumannii may help understand the mechanism behind the genetic mobilization and spread of AMR genes.

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