Metagenomic analysis of β-lactamase and carbapenemase genes in the wastewater resistome

Makowska, Nicoletta; Philips, Anna; Dabert, Mirosława; Nowis, Katarzyna; Trzebny, Artur; Koczura, Ryszard; Mokracka, Joanna

doi:10.1016/j.watres.2019.115277

Cited by 48 publications

(35 citation statements)

References 89 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In agreement with our results, previous studies reported an increase of ESBL and carbapenemaseproducing Enterobacteriaceae (CPE) in rivers and WWTPs, with high percentages of clinically relevant multidrug resistant bacteria and related genes (intI1, sul1, bla OXA , mcr-1, bla CTX-M15 , bla KPC and bla VIM , among others) that were still present in effluent samples, indicating an insufficient reduction during conventional wastewater treatment [47]. In this line, our results are in agreement with previous work published by Ojer-Usoz in the same region of Navarra [8], with a similar prevalence of ESBL after 6 years.…”

Section: Discussionsupporting

confidence: 93%

“…It is known that some ARB can be removed through conventional wastewater treatment processes [6], but still large numbers that survive in the effluent. Therefore, tertiary treatment methods or advanced treatment technologies are those that manage to eliminate some bacterial load and genes [47]. In this sense, UV and ozone-treatment have been investigated for a long time with the aim of reducing these microbial loads.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Multidrug-Resistant Bacteria Isolated from Different Aquatic Environments in the North of Spain and South of France

et al. 2020

View full text Add to dashboard Cite

Due to the global progress of antimicrobial resistance, the World Health Organization (WHO) published the list of the antibiotic-resistant “priority pathogens” in order to promote research and development of new antibiotics to the families of bacteria that cause severe and often deadly infections. In the framework of the One Health approach, the surveillance of these pathogens in different environments should be implemented in order to analyze their spread and the potential risk of transmission of antibiotic resistances by food and water. Therefore, the objective of this work was to determine the presence of high and critical priority pathogens included in the aforementioned list in different aquatic environments in the POCTEFA area (North Spain–South France). In addition to these pathogens, detection of colistin-resistant Enterobacteriaceae was included due its relevance as being the antibiotic of choice to treat infections caused by multidrug resistant bacteria (MDR). From the total of 80 analyzed samples, 100% of the wastewater treatment plants (WWTPs) and collectors (from hospitals and slaughterhouses) and 96.4% of the rivers, carried antibiotic resistant bacteria (ARB) against the tested antibiotics. Fifty-five (17.7%) of the isolates were identified as target microorganisms (high and critical priority pathogens of WHO list) and 58.2% (n = 32) of them came from WWTPs and collectors. Phenotypic and genotypic characterization showed that 96.4% were MDR and resistance to penicillins/cephalosporins was the most widespread. The presence of bla genes, KPC-type carbapenemases, mcr-1 and vanB genes has been confirmed. In summary, the presence of clinically relevant MDR bacteria in the studied aquatic environments demonstrates the need to improve surveillance and treatments of wastewaters from slaughterhouses, hospitals and WWTPs, in order to minimize the dispersion of resistance through the effluents of these areas.

show abstract

Section: Discussionsupporting

confidence: 93%

Section: Discussionmentioning

confidence: 99%

Multidrug-Resistant Bacteria Isolated from Different Aquatic Environments in the North of Spain and South of France

et al. 2020

View full text Add to dashboard Cite

show abstract

“…The eight capsular serotyping genes (K1, K2, K5, K20, K54, K57, K47, and K64) and pLVPKrelated genetic loci (rmpA2, terW, iutA, silS) were also determined by PCR and sequencing as previously described. [11][12][13][14][15][16] Outer Membrane Protein Isolation and SDS-PAGE Outer membrane proteins (OMPs) were isolated according to the rapid procedure of Carlone et al and separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) as previously described. 17 The reference strains K. pneumoniae ATCC 700603 and K. pneumoniae NUTH-K2044 were also used as quality control.…”

Section: Capsular Serotyping Resistance Genes and Plvpk-related Genmentioning

confidence: 99%

<p>Emergence of Hypervirulent Ceftazidime/Avibactam-Resistant <em>Klebsiella pneumoniae</em> Isolates in a Chinese Tertiary Hospital</p>

Liao

Huang

et al. 2020

IDR

View full text Add to dashboard Cite

Introduction: Carbapenem-resistant hypervirulent Klebsiella pneumoniae (CR-hvKP) is increasingly reported worldwide, but ceftazidime/avibactam (CAZ/AVI)-resistant hvKP isolates have rarely been observed. We attempted to characterize them in clinical CRKP isolates collected from a university hospital in China from March 2016 to March 2018. Methods: All isolates were analyzed by antimicrobial susceptibility testing, molecular detection of antibiotic resistance determinants, multilocus sequence typing (MLST), SDS-PAGE, and pulsed-field gel electrophoresis (PFGE). The pLVPK-related genetic loci (rmpA2, terW, iutA, and silS) were screened in all CAZ/AVI-resistant CRKP isolates for the presence of virulence plasmids by PCR. Capsule typing, serum killing assay, Galleria mellonella lethality experiments, and mouse lethality assay were conducted to identify CAZ/AVIresistant hvKP among isolates that carried all four virulence genes. Results: A total of 232 CRKP isolates were collected. Overall, CAZ/AVI-resistance was found in 8.2% (19/232) CRKP isolates isolated from patients with no history of previous CAZ/AVI-based treatment. Among these, 63.2% (12/19) were metallo-β-lactamaseproducing K. pneumoniae (MBL-KP), 52.6% (10/19) were Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae (KPC-KP), and 26.3% (5/19) produced both MBL and KPC. The presence of carbapenemase promoted a very high increase in CAZ/AVI minimum inhibitory concentration only when ompk35 and ompk36 were absent. Alarmingly, nine isolates had all four virulence genes for the presence of virulence plasmids. All nine isolates were considered to be CAZ/AVI-resistant hvKP according to the G. mellonella infection model and mouse lethality assay, with ST23 being the most common type (55.6%, 5/9). Conclusion: The newly emerged hypervirulent CAZ/AVI-resistant KP strain might cause a serious threat to public health, suggesting an urgent need for enhanced clinical awareness and epidemiologic surveillance.

show abstract

“…In addition to clinically isolated ARB, comparable ARB/ARGs have been detected from various environments around the world, including the e uent from wastewater treatment plants (WWTP) [4,5], lakes and rivers [6][7][8], and soil [9]. In particular, extended spectrum β-lactamase (ESBL)-producing E. coli or carbapenemase-producing organisms (CPOs) have been detected in rivers, lakes [10][11][12], and WWTP treated water [13][14][15] in several countries, posing a potential risk of infection, and Japan is no exception [16,17]. Moreover, hospital wastewater is the most ARB-contaminated source in WWTP [5,13,[18][19][20][21][22] and there is concern about the relationship between hospital wastewater and environmental pollution [23,24].…”

Section: Introductionmentioning

confidence: 99%

Metagenomic survey and whole genome sequencing of antimicrobial resistant bacteria in the sewage of a Japanese hospital

Katagiri

Kuroda

Sekizuka

et al. 2020

Preprint

View full text Add to dashboard Cite

Background The dissemination of antimicrobial-resistant bacteria (ARB) and the transfer of antimicrobial resistance genes (ARGs) are a threat to public health. Antibiotics are indispensable therapeutic agents essential for the treatment of infectious diseases; however, inappropriate use of antibiotics leads to the emergence of ARB. It is established that hospitals are closely involved in the spread of antimicrobial resistance (AMR), which impedes antibiotic treatment and subsequently increases mortality. In addition, excrement of patients or healthy carriers of ARB are discharged from the hospital sewage through the wastewater treatment plant (WWTP) into the rivers, causing an AMR burden on the environment.Method Metagenomic analysis was performed on the hospital sewage samples, followed by whole genome sequencing of the extended spectrum β-lactamase (ESBL)-producing organisms (EPOs). A comparative genome analysis was also performed between EPO isolates from sewage and clinical isolates.Results Metagenomic analysis showed that the hospital sewage tanks had bacterial flora corresponding to the human gut. During the study period, the hospital was relocated to a newly constructed building with new sewage tanks; however, the presence of ARB/ARGs in the new hospital sewage tanks became markedly equivalent to that of the old hospital within one month. The ESBL blaCTX−M and carbapenemase blaIMP genes were not much detected in the original hospital sewage samples by metagenome analysis, but selection on CHROMagar ESBL increased the sensitivity to detect those β-lactamase genes. Comparative genome analysis between sewage and clinical EPO isolates revealed partial similarity; however, most EPO isolates exhibited a notable difference (≥ 50) in single nucleotide variations based on core-genome phylogeny. This result suggests that only some of the sewage EPO isolates were originated from the clinical patient. Therapeutic agents in the hospital sewage were analyzed and the concentration of levofloxacin and clarithromycin was 0.0325 and 0.0135 µg/mL, respectively.Conclusions Whole genome analysis between sewage and clinical isolates suggested that healthy or asymptomatic carriers may be involved in the contamination of hospital sewage. Moreover, the hospital sewage tank may serve as a hotspot for the horizontal transfer of ARGs under the selective pressure of antimicrobial agents. Therefore, ARB monitoring in hospital sewage is expected to detect the presence of carriers prior to nosocomial ARB outbreaks. In addition, hospital wastewater should be treated suitably to bring ARB below detectable levels to reduce the environmental AMR burden.

show abstract

Metagenomic analysis of β-lactamase and carbapenemase genes in the wastewater resistome

Cited by 48 publications

References 89 publications

Multidrug-Resistant Bacteria Isolated from Different Aquatic Environments in the North of Spain and South of France

Multidrug-Resistant Bacteria Isolated from Different Aquatic Environments in the North of Spain and South of France

<p>Emergence of Hypervirulent Ceftazidime/Avibactam-Resistant <em>Klebsiella pneumoniae</em> Isolates in a Chinese Tertiary Hospital</p>

Metagenomic survey and whole genome sequencing of antimicrobial resistant bacteria in the sewage of a Japanese hospital

Contact Info

Product

Resources

About