The Resistome of ESKAPEE Pathogens in Untreated and Treated Wastewater: A Polish Case Study. - Related Documents




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275501.0000The Resistome of ESKAPEE Pathogens in Untreated and Treated Wastewater: A Polish Case Study. The aim of this study was to quantify ESKAPEE bacteria, genes encoding resistance to antibiotics targeting this group of pathogens, as well as integrase genes in municipal wastewater and river water. Environmental DNA was extracted from the collected samples and used in deep sequencing with the Illumina TruSeq kit. The abundance of bacterial genera and species belonging to the ESKAPEE group, 400 ARGs associated with this microbial group, and three classes of integrase genes were determined. A taxonomic analysis revealed that Acinetobacter was the dominant bacterial genus, whereas Acinetobacter baumannii and Escherichia coli were the dominant bacterial species. The analyzed samples were characterized by the highest concentrations of the following ARGs: bla(GES), bla(OXA-58), bla(TEM), qnrB, and qnrS. Acinetobacter baumannii, E. coli, and genes encoding resistance to β-lactams (bla(VEB-1), bla(IMP-1), bla(GES), bla(OXA-58), bla(CTX-M), and bla(TEM)) and fluoroquinolones (qnrS) were detected in samples of river water collected downstream from the wastewater discharge point. The correlation analysis revealed a strong relationship between A. baumannii (bacterial species regarded as an emerging human pathogen) and genes encoding resistance to all tested groups of antimicrobials. The transmission of the studied bacteria (in particular A. baumannii) and ARGs to the aquatic environment poses a public health risk.202236009054
275610.9999Characterization of carbapenem resistance in environmental samples and Acinetobacter spp. isolates from wastewater and river water in Poland. The aim of this study was to analyze the prevalence of carbapenem resistance genes in Acinetobacter spp. isolated from wastewater in a municipal WWTP and to determine their spread from treated wastewater to river water with the use of conventional and molecular microbiology methods (qualitative and quantitative PCR and metagenomic analysis). Samples of untreated and treated wastewater and samples of river water obtained upstream and downstream from the wastewater discharge point were collected in 3 seasons (February, June, and September) of 2019. Acinetobacter spp. isolates were obtained by the culture method on the CHROMagar™ Acinetobacter medium. Additionally, environmental DNA was extracted from the samples for metagenomic and qPCR analyses. The presence of beta-lactam resistance genes (Ambler class B and D), insertion sequence ISAba1, and class I, II, and III integron-integrase genes was determined, and the bacterial taxonomic structure and wastewater and river samples was analyzed. Out of the 301 isolates obtained on the CHROMagar™ Acinetobacter medium, 258 belonged to the genus Acinetobacter, including 21 isolates that were identified as Acinetobacter baumannii. The highest number of Acinetobacter spp. and A. baumannii isolates were obtained from wastewater and river water samples collected in June and September. The ISAba1/bla(OXA-51) complex was identified in 13 isolates, which confirms the occurrence of carbapenem-resistance isolates in the analyzed samples. The number of Acinetobacter isolates carrying antibiotic resistance genes (ARGs) increased in river water samples collected downstream from the wastewater discharge point (48 out of 258 isolates - 18.6%) compared to river water samples collected upstream from the wastewater discharge point (34 out of 258 isolates - 13.2%), which suggests that WWTP is a source of pollution in the natural environment. The conducted research provides evidence that bacteria of the genus Acinetobacter may spread alarming beta-lactam resistance in the environment and, therefore, pose a serious epidemiological threat.202235122847
275320.9999Extended-spectrum beta-lactamase-producing Escherichia coli and antimicrobial resistance in municipal and hospital wastewaters in Czech Republic: Culture-based and metagenomic approaches. Wastewaters serve as important hot spots for antimicrobial resistance and monitoring can be used to analyse the abundance and diversity of antimicrobial resistance genes at the level of large bacterial and human populations. In this study, whole genome sequencing of beta-lactamase-producing Escherichia coli and metagenomic analysis of whole-community DNA were used to characterize the occurrence of antimicrobial resistance in hospital, municipal and river waters in the city of Brno (Czech Republic). Cefotaxime-resistant E. coli were mainly extended-spectrum beta-lactamase (ESBL) producers (95.6%, n = 158), of which the majority carried bla(CTX-M) (98.7%; n = 151) and were detected in all water samples except the outflow from hospital wastewater treatment plant. A wide phylogenetic diversity was observed among the sequenced E. coli (n = 78) based on the detection of 40 sequence types and single nucleotide polymorphisms (average number 34,666 ± 15,710) between strains. The metagenomic analysis revealed a high occurrence of bacterial genera with potentially pathogenic members, including Pseudomonas, Escherichia, Klebsiella, Aeromonas, Enterobacter and Arcobacter (relative abundance >50%) in untreated hospital and municipal wastewaters and predominance of environmental bacteria in treated and river waters. Genes encoding resistance to aminoglycosides, beta-lactams, quinolones and macrolides were frequently detected, however bla(CTX-M) was not found in this dataset which may be affected by insufficient sequencing depth of the samples. The study pointed out municipal treated wastewater as a possible source of multi-drug resistant E. coli and antimicrobial resistance genes for surface waters. Moreover, the combination of two different approaches provided a more holistic view on antimicrobial resistance in water environments. The culture-based approach facilitated insight into the dynamics of ESBL-producing E. coli and the metagenomics shows abundance and diversity of bacteria and antimicrobial resistance genes vary across water sites.202133232750
531230.9999Presence and Persistence of ESKAPEE Bacteria before and after Hospital Wastewater Treatment. The metagenomic surveillance of antimicrobial resistance in wastewater has been suggested as a methodological tool to characterize the distribution, status, and trends of antibiotic-resistant bacteria. In this study, a cross-sectional collection of samples of hospital-associated raw and treated wastewater were obtained from February to March 2020. Shotgun metagenomic sequencing and bioinformatic analysis were performed to characterize bacterial abundance and antimicrobial resistance gene analysis. The main bacterial phyla found in all the samples were as follows: Proteobacteria, Bacteroides, Firmicutes, and Actinobacteria. At the species level, ESKAPEE bacteria such as E. coli relative abundance decreased between raw and treated wastewater, but S. aureus, A. baumannii, and P. aeruginosa increased, as did the persistence of K. pneumoniae in both raw and treated wastewater. A total of 172 different ARGs were detected; bla(OXA), bla(VEB), bla(KPC), bla(GES), mphE, mef, erm, msrE, AAC(6'), ant(3″), aadS, lnu, PBP-2, dfrA, vanA-G, tet, and sul were found at the highest abundance and persistence. This study demonstrates the ability of ESKAPEE bacteria to survive tertiary treatment processes of hospital wastewater, as well as the persistence of clinically important antimicrobial resistance genes that are spreading in the environment.202438930614
275440.9999Antimicrobial resistance in Aeromonas species isolated from aquatic environments in Brazil. AIM: The current study was conducted to determine the antimicrobial resistance profile and genetic relatedness of Aeromonas sp. isolated from healthcare and urban effluents, wastewater treatment plant (WWTP) and river water. METHODS AND RESULTS: We detected the presence of genes conferring resistance to β-lactam, quinolone and aminoglycoside. Multilocus sequence typing was carried out to differentiate the strains, and multilocus phylogenetic analysis was used to identify the species. A total of 28 cefotaxime-resistant Aeromonas sp. strains were identified, harbouring uncommon Guiana-extended-spectrum (GES)-type β-lactamases (GES-1, GES-5, GES-7 and GES-16). Multidrug-resistant Aeromonas sp. were found in hospital wastewater, WWTP and sanitary effluent, and A. caviae was identified as the most prevalent species (85·7%). CONCLUSION: The release of untreated healthcare effluents, presence of antimicrobials in the environment, in addition to multidrug-resistant Aeromonas sp., are all potential factors for the spread of resistance. SIGNIFICANCE AND IMPACT OF THE STUDY: We identified a vast repertoire of antimicrobial resistance genes (ARG) in Aeromonas sp. from diverse aquatic ecosystems, including those that encode enzymes degrading broad-spectrum antimicrobials widely used to treat healthcare-associated infections. Hospital and sanitary effluents serve as potential sources of bacteria harbouring ARG and are a threat to public health.202133306232
275250.9999Antibiotic Resistance in Enterobacteriaceae from Surface Waters in Urban Brazil Highlights the Risks of Poor Sanitation. Surface waters are an unappreciated reservoir of antimicrobial resistance (AMR). Poor sanitation brings different species of environmental bacteria into contact, facilitating horizontal gene transfer. To investigate the role of surface waters as potential reservoirs of AMR, we studied the point prevalence of fecal contamination, AMR genes, and Enterobacteriaceae in an urban lake and rural river system in Northeast Brazil in comparison with a lake and sewer system in Northeast Ohio in the United States. Surface water samples were examined for evidence of human fecal contamination using microbial source tracking and screened for plasmid-mediated fluoroquinolone resistance and carbapenemase genes. Enterobacteriaceae were detected using selective agar followed by antimicrobial susceptibility testing and detection of AMR genes by microarray, and classified by repetitive sequence-based polymerase chain reaction and multilocus sequence typing. Concentrations of human fecal bacteria in the Brazilian urban lake and sewage in Northeast Ohio were similarly high. Filtered water samples from the Brazilian urban lake, however, showed the presence of bla (OXA-48), bla (KPC), bla (VIM-2), qnrS, and aac(6')-lb-cr, whereas only bla (VIM-2) was identified in raw sewage from Northeast Ohio. From the Brazilian urban lake, 85% of the Enterobacteriaceae (n = 40) cultured were resistant to at least one clinically important antibiotic, including ST131 Escherichia coli harboring the extended-spectrum beta-lactamase CTX-M. Although two isolates demonstrated polymyxin resistance, mcr-1/2 was not detected. Our findings indicate that surface waters in an urban Brazilian site can serve as an environmental reservoir of AMR and that improving wastewater treatment and sanitation generally may ameliorate AMR dissemination.201930994094
275160.9999Recovery of clinically relevant multidrug-resistant Klebsiella pneumoniae lineages from wastewater in Kumasi Metropolis, Ghana. Antimicrobial resistance (AMR) is under-monitored in Africa, with few reports characterizing resistant bacteria from the environment. This study examined physicochemical parameters, chemical contaminants and antibiotic-resistant bacteria in waste stabilization pond effluents, hospital wastewater and domestic wastewater from four sewerage sites in Kumasi. The bacteria isolates were sequenced. Three sites exceeded national guidelines for total suspended solids, biochemical oxygen demand, chemical oxygen demand and electrical conductivity. Although sulfamethoxazole levels were low, the antibiotic was detected at all sites. Multi-drug-resistant Klebsiella pneumoniae and Pseudomonas aeruginosa were isolated with multi-locus sequence typing identifying K. pneumoniae strains as ST18 and ST147, and P. aeruginosa as ST235, all of clinical relevance. A comparison of ST147 genomes with isolates from human infections in Africa showed remarkable similarity and shared AMR profiles. Thirteen of the twenty-one plasmids from ST147 harbored at least one AMR gene, including blaCTX-M-15 linked to copper-resistance genes. Our study demonstrated high bacterial counts and organic matter in the analysed wastewater. The recovery of clinically significant isolates with multiple antibiotic and heavy metal resistance genes from the wastewater samples raises public health concerns.202439516432
273470.9999High Frequency of Antibiotic Resistance Genes (ARGs) in the Lerma River Basin, Mexico. The spread of beta-lactamase-producing bacteria is of great concern and the environment has been found to be a main source of contamination. Herein, it was proposed to determine the frequency of antimicrobial-resistant-Gram-negative bacteria throughout the Lerma River basin using phenotypic and molecular methods. Resistant bacteria were isolated with chromogenic media and antimicrobial susceptibility tests were used to characterize their resistance. ARGs for beta-lactams, aminoglycosides, and quinolones were detected by PCR. Species were identified by Sanger sequencing the 16S rRNA gene and the representative genomes of MDR strains were sequenced by NGS. A high variation in the number of isolates was observed in the 20 sampled sites, while observing a low diversity among the resistant bacteria. Of the 12 identified bacterial groups, C. freundii, E. coli, and S. marcescens were more predominant. A high frequency of resistance to beta-lactams, quinolones, and aminoglycosides was evidenced, where the bla(CTX,)qnrB, qnrS y, and aac(6')lb-cr genes were the most prevalent. C. freundii showed the highest frequency of MDR strains. Whole genome sequencing revealed that S. marcescens and K. pneumoniae showed a high number of shared virulence and antimicrobial resistance genes, while E. coli showed the highest number of unique genes. The contamination of the Lerma River with MDR strains carrying various ARGs should raise awareness among environmental authorities to assess the risks and regulations regarding the optimal hygienic and sanitary conditions for this important river that supports economic activities in the different communities in Mexico.202236360888
277980.9998Antibiotic resistance determinants among carbapenemase producing bacteria isolated from wastewaters of Kathmandu, Nepal. The emergence of carbapenem resistant bacteria (CRB) possesses a remarkable threat to the health of humans. CRB and carbapenem resistance genes (CRGs) have frequently been reported in clinical isolates from hospitals, however, their occurrence and distribution in wastewaters from various sources and river water have not been emphasized in Nepal. So, this study aimed to detect carbapenem resistant bacterial isolates and their resistance determinants in river water and different types of wastewaters. River water and both untreated and treated wastewater samples from hospitals, pharmaceutical industries, and municipal sewage were collected in summer and winter seasons. From 68 grab wastewater samples, CRB were detected only in 16 samples, which included eight hospital wastewater, and four each from untreated municipal sewage and river water. A total of 25 CRB isolates were detected with dominance of E. coli (44.0%) and K. pneumoniae (24.0%). The majority of the isolates harbored bla(NDM-1) (76.0%), followed by bla(OXA) (36.0%) and bla(KPC) (20.0%) genes. Hospital wastewater majorly contributed to the presence of bla(NDM-1), bla(KPC), and bla(OXA) along with intI1 genes compared to river water and untreated municipal sewage, especially during the winter season. However, CRB were not detected in treated effluents of hospitals and municipal sewage, and both influents and effluents from pharmaceutical industries. The combined presence of each bla(NDM-1) & bla(OXA) and bla(KPC) & bla(OXA) occurred in 16.0% of the bacterial isolates. The increased minimum inhibitory concentration (MIC) of meropenem was significantly associated with the presence of CRGs. The results of this study highlight the significance of carbapenem resistance in bacteria isolated from wastewater and river water, and underscore the necessity for efficient monitoring and control strategies to prevent the dispersion of carbapenem resistance in the environment and its potential consequences on human health.202438114055
273690.9998Characterization of Bacterial Communities and Their Antibiotic Resistance Profiles in Wastewaters Obtained from Pharmaceutical Facilities in Lagos and Ogun States, Nigeria. In Nigeria, pharmaceutical wastewaters are routinely disseminated in river waters; this could be associated with public health risk to humans and animals. In this study, we characterized antibiotic resistant bacteria (ARB) and their antibiotic resistance profile as well as screening for sul1 and sul2 genes in pharmaceutical wastewater effluents. Bacterial composition of the wastewater sources was isolated on non-selective media and characterized by the polymerase chain reaction (PCR) amplification of the 16S rRNA genes, with subsequent grouping using restriction fragment length polymorphism (RFLP) and sequencing. The antibiotics sensitivity profiles were investigated using the standard disk diffusion plate method and the minimum inhibitory concentrations (MICs) of selected antibiotics on the bacterial isolates. A total of 254 bacterial strains were isolated, and majority of the isolates were identified as Acinetobacter sp., Klebsiella pneumonia, Proteus mirabilis, Enterobacter sp. and Bacillus sp. A total of 218 (85.8%) of the bacterial isolates were multidrug resistant. High MICs values were observed for all antibiotics used in the study. The result showed that 31.7%, 21.7% and 43.3% of the bacterial isolates harbored sul1, sul2, and Intl1 genes, respectively. Pharmaceuticals wastewaters are potential reservoirs of ARBs which may harbor resistance genes with possible risk to public health.201829966226
3313100.9998The Prevalence and Characterization of Extended-Spectrum β-Lactamase- and Carbapenemase-Producing Bacteria from Hospital Sewage, Treated Effluents and Receiving Rivers. Hospital sewage plays a key role in the dissemination of antibiotic-resistant genes (ARGs) by serving as an environmental antimicrobial resistance reservoir. In this study, we aimed to characterize the cephalosporin- and carbapenem-resistant isolates from hospital sewage and receiving rivers. The results showed that ESBL (bla(CTX-M)) and carbapenemase genes (bla(NDM) and bla(KPC)) were widely detected in a number of different bacterial species. These resistance genes were mainly harbored in Enterobacteriaceae, followed by Acinetobacter and Aeromonas isolates. More attention should be given to these bacteria as important vectors of ARGs in the environment. Furthermore, we showed that the multidrug resistance phenotype was highly prevalent, which was found in 85.5% Enterobacteriaceae and 75% Acinetobacter strains. Notably, the presence of carbapenemase genes in isolates from treated effluents and receiving rivers indicates that the discharges of wastewater treatment plants could be an important source for high-risk resistance genes propagation to the environment. In conclusion, this study shows a high prevalence of ESBL- and carbapenemase-producing bacteria in hospital sewage and receiving rivers in China. These findings have serious implications for human health, and also suggest the need for more efforts to control the dissemination of resistant bacteria from hospital sewage into the environment.202032069792
1805110.9998The prevalence of multidrug-resistant Aeromonas spp. in the municipal wastewater system and their dissemination in the environment. The objective of this study was to identify the determinants of antibiotic resistance and virulence of Aeromonas spp. isolated from treated wastewater (WW) and from samples of river water collected upstream (URW) and downstream (DRW) from the effluent discharge point. The resistance of Aeromonas spp. to antibiotics that are widely used in human and veterinary medicine, including beta-lactams, tetracyclines, glycylcyclines, fluoroquinolones, aminoglycosides and sulfamethoxazole-trimethoprim, was analyzed by disk diffusion method. The prevalence of hemolysins, aerolysins (virulence factors) and integrase genes was determined. A total of 83 Aeromonas spp. strains were selected from the isolates obtained from river water and wastewater samples. The highest percentage (81.8%) of multidrug-resistant isolates was noted in DRW samples. The analyzed isolates were most frequently resistant to beta-lactams, tetracyclines and aminoglycosides, whereas resistance to glycylcyclines was least common. Isolates resistant to beta-lactams most frequently harbored bla(TEM) and bla(OXA) genes. The group of genes encoding resistance to tetracyclines was most frequently represented by tet(E) and tet(O). Genes encoding virulence ahh1 (heat-labile hemolysin) or integrase intI1 occurred more frequently in the strains isolated from DRW than URW. An analysis of genetic relatedness revealed two main clusters - one with closely related WW and DRW isolates and one with less related isolates from all analyzed samples. The results of this study indicate that regardless of the applied treatment, municipal sewage may be a reservoir of antibiotic-resistant bacteria, antibiotic resistance and virulence genes and that treated water can contaminate other environmental domains.201829353783
2738120.9998Diversity of bacteria carrying antibiotic resistance genes in hospital raw sewage in Southeastern Brazil. In recent decades, antibiotic-resistant bacteria (ARB) emerged and spread among humans and animals worldwide. In this study, we evaluated the presence of ARB and antibiotic resistance genes (ARGs) in the raw sewage of two hospitals in Brazil. Sewage aliquots were inoculated in a selective medium with antibiotics. Bacterial identification was performed by MALDI-TOF and ARGs were assessed by polymerase chain reaction (PCR). A total of 208 strains from both hospitals were isolated (H1 = 117; H2 = 91). A wide variety of Enterobacterales and non-Enterobacterales species were isolated and most of them were Enterobacter spp. (13.0%), Proteus mirabilis (10.1%), and Klebsiella pneumoniae (9.6%). blaTEM and blaKPC were the most frequent β-lactamase-encoding genes and the predominant macrolide resistance genes were mph(A) and mel. Many species had the three tetracycline resistance genes (tetD, tetM, tetA) and strB was the prevalent aminoglycoside resistance gene. Two Staphylococcus haemolyticus strains had the mecA gene. Quinolone, colistin, and vancomycin resistance genes were not found. This study showed that hospital raw sewage is a great ARB and ARG disseminator. Strict monitoring of hospital sewage treatment is needed to avoid the spread of these genes among bacteria in the environment.202336640035
2737130.9998Meropenem-resistant bacteria in hospital effluents in Seoul, Korea. This study aimed to understand the prevalence, diversity, antibiotic resistance, β-lactamase gene types, and possibility of environmental survival of meropenem-resistant bacteria present in hospital effluents in Seoul, Korea. Water sampling was performed at five general hospitals in Seoul, Korea, in January 2017. Water samples were plated in triplicate on tryptic soy agar plates with 16 mg/L meropenem. Meropenem-resistant bacteria were selected and subjected to 16S rRNA analysis for species determination and PCR for identification of β-lactamase gene types. Resistant bacteria were cultured in sterilized surface water. Meropenem-resistant bacteria exhibited resistance to more than 12 antibiotics and possessed several β-lactamase genes, such as those encoding OXT-M, NDM-1, AmpC, and OXA. They were able to multiply and survive in sterilized surface water for up to 60 days. Multidrug-resistant bacteria represent an environmental health risk, as they can survive in the environment for an extended period of time. Therefore, these bacteria should be monitored before discharge.201830361772
1810140.9998Characterization of Metagenomes in Urban Aquatic Compartments Reveals High Prevalence of Clinically Relevant Antibiotic Resistance Genes in Wastewaters. The dissemination of antimicrobial resistance (AMR) is an escalating problem and a threat to public health. Comparative metagenomics was used to investigate the occurrence of antibiotic resistant genes (ARGs) in wastewater and urban surface water environments in Singapore. Hospital and municipal wastewater (n = 6) were found to have higher diversity and average abundance of ARGs (303 ARG subtypes, 197,816 x/Gb) compared to treated wastewater effluent (n = 2, 58 ARG subtypes, 2,692 x/Gb) and surface water (n = 5, 35 subtypes, 7,985 x/Gb). A cluster analysis showed that the taxonomic composition of wastewaters was highly similar and had a bacterial community composition enriched in gut bacteria (Bacteroides, Faecalibacterium, Bifidobacterium, Blautia, Roseburia, Ruminococcus), the Enterobacteriaceae group (Klebsiella, Aeromonas, Enterobacter) and opportunistic pathogens (Prevotella, Comamonas, Neisseria). Wastewater, treated effluents and surface waters had a shared resistome of 21 ARGs encoding multidrug resistant efflux pumps or resistance to aminoglycoside, macrolide-lincosamide-streptogramins (MLS), quinolones, sulfonamide, and tetracycline resistance which suggests that these genes are wide spread across different environments. Wastewater had a distinctively higher average abundance of clinically relevant, class A beta-lactamase resistant genes (i.e., bla(KPC), bla(CTX-M), bla(SHV), bla(TEM)). The wastewaters from clinical isolation wards, in particular, had a exceedingly high levels of bla(KPC-2) genes (142,200 x/Gb), encoding for carbapenem resistance. Assembled scaffolds (16 and 30 kbp) from isolation ward wastewater samples indicated this gene was located on a Tn3-based transposon (Tn4401), a mobilization element found in Klebsiella pneumonia plasmids. In the longer scaffold, transposable elements were flanked by a toxin-antitoxin (TA) system and other metal resistant genes that likely increase the persistence, fitness and propagation of the plasmid in the bacterial host under conditions of stress. A few bacterial species (Enterobacter cloacae, Klebsiella pneumoniae, Citrobacter freundii, Pseudomonas aeruginosa) that were cultured from the isolation ward wastewaters on CHROMagar media harbored the bla(KPC-2) gene. This suggests that hospital wastewaters derived from clinical specialty wards are hotspots for the spread of AMR. Assembled scaffolds of other mobile genetic elements such as IncQ and IncF plasmids bearing quinolone resistance genes (qnrS1, qnrS2) and the class A beta-lactamase gene (bla(TEM-1)) were recovered in wastewater samples which may aid the transfer of AMR.201729201017
2733150.9998Prevalence and diversity of carbapenem-resistant bacteria in untreated drinking water in Portugal. We examined the prevalence and diversity of carbapenem-resistant bacteria (CRB) in untreated drinking water. Prevalence was estimated in plate count agar (PCA) and R2A media with or without antibiotics. Clonal relatedness of isolates was established by repetitive extragenic palindroitic (REP)-PCR. Phylogeny was based on the 16S rRNA gene. Antimicrobial susceptibility was assessed by disc diffusion methods. Genes encoding beta-lactamases and integrases were inspected by PCR. CRB ranged from 0.02% to 15.9% of cultivable bacteria, while ampicillin-resistant bacteria ranged from 1.5% to 31.4%. Carbapenem-resistant isolates affiliated with genera Stenotrophomonas, Pseudomonas, Janthinobacterium, Chryseobacterium, Sphingobacterium, Acidovorax, Caulobacter, Cupriavidus, and Sphingomonas. CRB were highly resistant to beta-lactams, but mostly susceptible to other classes. Transmissible beta-lactamase genes and integrase genes were not detected. The genus-specific bla(L1) was detected in 61% of the Stenotrophomonas isolates. Contrarily to what has been reported for extensively used antibiotics, low levels of carbapenem resistance were detected in untreated drinking water, often represented by intrinsically resistant genera. Production of chromosomal-encoded carbapenemases was the prevalent carbapenem resistance mechanism. Results suggest that the dissemination of anthropogenic-derived carbapenem resistance is at an early stage. This presents an opportunity to rationally develop monitoring strategies to identify dissemination routes and assess the impact of human actions in the environmental resistome.201222663561
2748160.9998Multiresistance, beta-lactamase-encoding genes and bacterial diversity in hospital wastewater in Rio de Janeiro, Brazil. AIMS: To investigate the bacterial diversity, antimicrobial resistance patterns and types of beta-lactamase genes in Gram-negative bacteria isolated from a hospital sewage treatment plant. METHODS AND RESULTS: Between July and December 2008, we collected samples from influent, clarifier tank effluent and chlorine contact tank effluent from a sewage treatment plant service of a hospital located in the city of Rio de Janeiro, Brazil. Of the 221 isolates identified, 40% were characterized as extended-spectrum beta-lactamase (ESBL) producers. Nonpathogenic micro-organisms and some pathogenic genera were quantified. The most common ESBL-producing isolates were Klebsiella pneumoniae, Enterobacter cloacae and Escherichia coli. The bla(TEM), bla(SHV) and bla(CTX-M) genes were detected in 82, 48 and 67% of bacterial isolates, respectively. CONCLUSIONS: Results showed that hospital wastewater treatment plant is not suitable systems for the removal of all antibiotic-resistant micro-organisms present in hospital wastewaters. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides evidence that bacteria resistant to multiple antibiotics and their resistance genes that are usually present in the hospital can reach the environment, even after the use of hospital wastewater treatment plants.201121672095
3319170.9998Extended-spectrum beta-lactamase (ESBL)-positive Enterobacteriaceae in municipal sewage and their emission to the environment. The spread of Gram-negative bacteria with plasmid-borne extended-spectrum beta-lactamases (ESBLs) has become a worldwide problem. Their prevalence is increasing, both in hospitals and in the environment. The aim of this study was to investigate the presence of ESBL-positive Enterobacteriaceae in municipal sewage and their emission to the ambient air and the river receiving effluent from wastewater treatment plant (WWTP). In the group of 455 isolated strains, up to 19.8% (90 isolates) were phenotypic ESBL-producers. They were detected in the 63 (100%) of sewage samples analyzed, 7 (33.3%) of river water and in 10 (23.8%) of air samples collected at the WWTP area. The plasmid-mediated genes encoding beta-lactams resistance were detected in almost 10% out of bacteria of the WWTP's final effluents and in above 32% out of bacteria of air at the WWTP area. It confirms that those genes are released into the environment, which might facilitate further dissemination among environmental bacteria. Moreover, genes encoding antibiotic resistance were shown to be transferrable to an Escherichia coli recipient strain, which indicates a high possibility of horizontal gene transfer among strains of different genera within the sewage and environmental samples. This study demonstrated that despite the treatment, the municipal sewage may be a reservoir of antibiotic-resistant microorganisms and plasmid-mediated antibiotic resistance genes. This may pose a public health risk, which requires future evaluation and control.201323886578
2735180.9998Insight into the Antibiotic Resistance of Bacteria Isolated from Popular Aquatic Products Collected in Zhejiang, China. The present study was aimed to obtain a close insight into the distribution and diversity of antibiotic-resistant bacteria (ARB) and antibiotic-resistance genes (ARGs) among the aquatic products collected in Zhejiang, China. A total of 136 presumptive ARB picked up from six aquatic samples were classified into 22 genera and 49 species based on the 16S rDNA sequencing. Aeromonas spp., Shewanella spp., Acinetobacter spp., Myroides spp., Pseudomonas spp., and Citrobacter spp. accounted for 80% of the ARB. Among them, 109 isolates (80.15%) exhibited resistance to at least one antibiotic. Most isolates showed resistance to not only the originally selected drug but also to one to three other tested drugs. The diversity of ARB distributed in different aquatic products was significant. Furthermore, the resistance data obtained from genotypic tests were not entirely consistent with the results of the phenotypic evaluation. The genes qnrS, tetA, floR, and cmlA were frequently detected in their corresponding phenotypic resistant isolates. In contrast, the genes sul2, aac(6')-Ib, and bla (PSE) were less frequently found in the corresponding phenotypically resistant strains. The high diversity and detection rate of ARB and ARGs in aquaculture might be a significant threat to the food chains closely related to human health.202336929890
5314190.9998High prevalence of colistin resistance genes in German municipal wastewater. Bacterial resistance against the last-resort antibiotic colistin is of increasing concern on a global scale. Wastewater is suspected to be one of the pathways by which resistant bacteria and the respective genes are disseminated. We employed a metagenomics approach to detect and quantify colistin resistance genes in raw municipal wastewater sampled at 9 locations all over Germany (14 samples in total, collected in 2016/2017). Our data support the findings of earlier studies according to which the prevalence of the colistin resistance gene mcr-1 is still low. However, we were able to demonstrate that the total prevalence of colistin resistance genes is dramatically underestimated if the focus is put on that specific gene alone. In comparison to mcr-1, other gene variants like mcr-3 and mcr-7 proved to be 10 to 100 times more abundant in samples of untreated wastewater. The average relative abundances expressed as copies per 16S rRNA gene copies were 2.3×10(-3) for mcr-3, 2.2×10(-4) for mcr-4, 3.0×10(-4) for mcr-5, and 4.4×10(-4) for mcr-7. While these four gene variants were ubiquitous in all 14 samples, mcr-1 was detected only once at a relative abundance of 1.4×10(-5). Our results suggest a high risk of increasing incidence of colistin resistance as large amounts of mcr genes are continuously disseminated to diverse microbial communities via the wastewater path.201931398645