Repeated Isolation of Extended-Spectrum-β-Lactamase-Positive Escherichia coli Sequence Types 648 and 131 from Community Wastewater Indicates that Sewage Systems Are Important Sources of Emerging Clones of Antibiotic-Resistant Bacteria

ABSTRACT Antibiotic resistance in bacteria is an emerging problem globally. Resistant bacteria are found in human and animal microbiota, as well as in the environment. Wastewater receives bacteria from all these sources and thus can provide a measurement of abundance and diversity of antibiotic-resistant bacteria circulating in communities. In this study, water samples were collected from a wastewater pump station in a Norwegian suburban community over a period of 15 months. A total of 45 daily samples were cultured and analyzed for the presence of Escherichia coli. Eighty E. coli-like colonies were collected from each daily sample and then phenotyped and analyzed for antibiotic resistance using the PhenePlate-AREB system. During the sampling period, two unique E. coli phenotypes with resistance to cefotaxime and cefpodoxime indicating carriage of extended-spectrum β-lactamases (ESBL) were observed repeatedly. Whole-genome sequencing of 15 representative isolates from the two phenotypes identified these as two distinct clones belonging to the two globally spread E. coli multilocus sequence types (STs) ST131 and ST648 and carrying blaCTX-M-15. The number of ESBL-positive E. coli strains in the community wastewater pump station was 314 of 3,123 (10%) analyzed E. coli strains. Of the ESBL-positive isolates, 37% belonged to ST648, and 7% belonged to ST131. Repeated findings of CTX-M-15-positive ST648 and ST131 over time indicate that these STs are resident in the analyzed wastewater systems and/or circulate abundantly in the community.

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Prevalence and Characterization of Extended-Spectrum β-Lactamase-Producing Antibiotic-Resistant Escherichia coli and Klebsiella pneumoniae in Ready-to-Eat Street Foods

Antibiotics ◽

10.3390/antibiotics10070850 ◽

2021 ◽

Vol 10 (7) ◽

pp. 850

Author(s):

Shobha Giri ◽

Vaishnavi Kudva ◽

Kalidas Shetty ◽

Veena Shetty

Keyword(s):

Escherichia Coli ◽

Food Safety ◽

Klebsiella Pneumoniae ◽

Rural Areas ◽

Significant Risk ◽

Microbiological Quality ◽

Resistant Bacteria ◽

Antibiotic Resistant ◽

E Coli ◽

Extended Spectrum

As the global urban populations increase with rapid migration from rural areas, ready-to-eat (RTE) street foods are posing food safety challenges where street foods are prepared with less structured food safety guidelines in small and roadside outlets. The increased presence of extended-spectrum-β-lactamase (ESBL) producing bacteria in street foods is a significant risk for human health because of its epidemiological significance. Escherichia coli and Klebsiella pneumoniae have become important and dangerous foodborne pathogens globally for their relevance to antibiotic resistance. The present study was undertaken to evaluate the potential burden of antibiotic-resistant E. coli and K. pneumoniae contaminating RTE street foods and to assess the microbiological quality of foods in a typical emerging and growing urban suburb of India where RTE street foods are rapidly establishing with public health implications. A total of 100 RTE food samples were collected of which, 22.88% were E. coli and 27.12% K. pneumoniae. The prevalence of ESBL-producing E. coli and K. pneumoniae was 25.42%, isolated mostly from chutneys, salads, paani puri, and chicken. Antimicrobial resistance was observed towards cefepime (72.9%), imipenem (55.9%), cefotaxime (52.5%), and meropenem (16.9%) with 86.44% of the isolates with MAR index above 0.22. Among β-lactamase encoding genes, blaTEM (40.68%) was the most prevalent followed by blaCTX (32.20%) and blaSHV (10.17%). blaNDM gene was detected in 20.34% of the isolates. This study indicated that contaminated RTE street foods present health risks to consumers and there is a high potential of transferring multi-drug-resistant bacteria from foods to humans and from person to person as pathogens or as commensal residents of the human gut leading to challenges for subsequent therapeutic treatments.

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Comparison of Antibiotic Resistance Profile of Escherichia coli between Pristine and Human-Impacted Sites in a River

Antibiotics ◽

10.3390/antibiotics10050575 ◽

2021 ◽

Vol 10 (5) ◽

pp. 575

Author(s):

Emi Nishimura ◽

Masateru Nishiyama ◽

Kei Nukazawa ◽

Yoshihiro Suzuki

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Resistant Bacteria ◽

Downstream Site ◽

Antibiotic Resistant Bacteria ◽

Antibiotic Resistant ◽

Upstream Site ◽

Resistance Profile ◽

Antibiotic Resistance Profile ◽

Significant Difference

Information on the actual existence of antibiotic-resistant bacteria in rivers where sewage, urban wastewater, and livestock wastewater do not load is essential to prevent the spread of antibiotic-resistant bacteria in water environments. This study compared the antibiotic resistance profile of Escherichia coli upstream and downstream of human habitation. The survey was conducted in the summer, winter, and spring seasons. Resistance to one or more antibiotics at upstream and downstream sites was on average 18% and 20%, respectively, and no significant difference was observed between the survey sites. The resistance rates at the upstream site (total of 98 isolated strains) to each antibiotic were cefazolin 17%, tetracycline 12%, and ampicillin 8%, in descending order. Conversely, for the downstream site (total of 89 isolated strains), the rates were ampicillin 16%, cefazolin 16%, and tetracycline 1% in descending order. The resistance rate of tetracycline in the downstream site was significantly lower than that of the upstream site. Furthermore, phylogenetic analysis revealed that many strains showed different resistance profiles even in the same cluster of the Pulsed-Field Gel Electrophoresis (PFGE) pattern. Moreover, the resistance profiles differed in the same cluster of the upstream and the downstream sites. In flowing from the upstream to the downstream site, it is plausible that E. coli transmitted or lacked the antibiotic resistance gene.

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ESKAPE Bacteria and Extended-Spectrum-β-Lactamase-Producing Escherichia coli Isolated from Wastewater and Process Water from German Poultry Slaughterhouses

Applied and Environmental Microbiology ◽

10.1128/aem.02748-19 ◽

2020 ◽

Vol 86 (8) ◽

Cited By ~ 7

Author(s):

Mykhailo Savin ◽

Gabriele Bierbaum ◽

Jens Andre Hammerl ◽

Céline Heinemann ◽

Marijo Parcina ◽

...

Keyword(s):

Escherichia Coli ◽

Wastewater Treatment ◽

Resistance Genes ◽

Wastewater Treatment Plants ◽

Process Water ◽

Resistant Bacteria ◽

Content Type ◽

E Coli ◽

Extended Spectrum ◽

Clinically Relevant Bacteria

ABSTRACT The wastewater of livestock slaughterhouses is considered a source of antimicrobial-resistant bacteria with clinical relevance and may thus be important for their dissemination into the environment. To get an overview of their occurrence and characteristics, we investigated process water (n = 50) from delivery and unclean areas as well as wastewater (n = 32) from the in-house wastewater treatment plants (WWTPs) of two German poultry slaughterhouses (slaughterhouses S1 and S2). The samples were screened for ESKAPE bacteria (Enterococcus spp., Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter spp.) and Escherichia coli. Their antimicrobial resistance phenotypes and the presence of extended-spectrum-β-lactamase (ESBL), carbapenemase, and mobilizable colistin resistance genes were determined. Selected ESKAPE bacteria were epidemiologically classified using different molecular typing techniques. At least one of the target species was detected in 87.5% (n = 28/32) of the wastewater samples and 86.0% (n = 43/50) of the process water samples. The vast majority of the recovered isolates (94.9%, n = 448/472) was represented by E. coli (39.4%), the A. calcoaceticus-A. baumannii (ACB) complex (32.4%), S. aureus (12.3%), and K. pneumoniae (10.8%), which were widely distributed in the delivery and unclean areas of the individual slaughterhouses, including their wastewater effluents. Enterobacter spp., Enterococcus spp., and P. aeruginosa were less abundant and made up 5.1% of the isolates. Phenotypic and genotypic analyses revealed that the recovered isolates exhibited diverse resistance phenotypes and β-lactamase genes. In conclusion, wastewater effluents from the investigated poultry slaughterhouses exhibited clinically relevant bacteria (E. coli, methicillin-resistant S. aureus, K. pneumoniae, and species of the ACB and Enterobacter cloacae complexes) that contribute to the dissemination of clinically relevant resistances (i.e., blaCTX-M or blaSHV and mcr-1) in the environment. IMPORTANCE Bacteria from livestock may be opportunistic pathogens and carriers of clinically relevant resistance genes, as many antimicrobials are used in both veterinary and human medicine. They may be released into the environment from wastewater treatment plants (WWTPs), which are influenced by wastewater from slaughterhouses, thereby endangering public health. Moreover, process water that accumulates during the slaughtering of poultry is an important reservoir for livestock-associated multidrug-resistant bacteria and may serve as a vector of transmission to occupationally exposed slaughterhouse employees. Mitigation solutions aimed at the reduction of the bacterial discharge into the production water circuit as well as interventions against their further transmission and dissemination need to be elaborated. Furthermore, the efficacy of in-house WWTPs needs to be questioned. Reliable data on the occurrence and diversity of clinically relevant bacteria within the slaughtering production chain and in the WWTP effluents in Germany will help to assess their impact on public and environmental health.

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Multiple NDM-5-Expressing Escherichia Coli Isolates From an Immunocompromised Pediatric Host

Open Forum Infectious Diseases ◽

10.1093/ofid/ofaa018 ◽

2020 ◽

Vol 7 (2) ◽

Cited By ~ 2

Author(s):

Tim Flerlage ◽

Jessica N Brazelton de Cardenas ◽

Cherilyn D Garner ◽

Nur A Hasan ◽

Hiren Karathia ◽

...

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Antibiotic Resistance Gene ◽

The United States ◽

Multidrug Resistant ◽

Resistant Bacteria ◽

Invasive Infection ◽

Anticancer Chemotherapy ◽

E Coli ◽

Sequence Types

Abstract Background Genes conferring carbapenem resistance have disseminated worldwide among Gram-negative bacteria. Here we present longitudinal changes in clinically obtained Escherichia coli isolates from 1 immunocompromised pediatric patient. This report demonstrates potential for antibiotic resistance genes and plasmids to emerge over time in clinical isolates from patients receiving intensive anticancer chemotherapy and broad-spectrum antibiotics. Methods Thirty-three isolates obtained over 7 months from 1 patient were included. Clinical data were abstracted from the medical record. For each isolate, studies included phenotypic antibacterial resistance patterns, sequence typing, bacterial isolate sequencing, plasmid identification, and antibiotic resistance gene identification. Results Sites of isolation included blood, wound culture, and culture for surveillance purposes from the perianal area. Isolates were of 5 sequence types (STs). All were resistant to multiple classes of antibiotics; 23 (69.6%) were phenotypically resistant to all carbapenems. The blaNDM-5 gene was identified in 22 (67%) isolates, all of ST-167 and ST-940, and appeared to coincide with the presence of the IncFII and IncX3 plasmid. Conclusions We present unique microbiologic data from 33 multidrug-resistant E. coli isolates obtained over the course of 7 months from an individual patient in the United States. Two E. coli sequence types causing invasive infection in the same patient and harboring the blaNDM-5 gene, encoded on the IncX3 plasmid and the IncFII plasmid, were identified. This study highlights the emergence of multidrug-resistant bacteria on antibiotic therapy and the necessity of adequate neutrophil number and function in the clearance of bacteremia.

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The occurrence of antibiotic-resistant bacteria, including Escherichia coli, in municipal wastewater and river water

E3S Web of Conferences ◽

10.1051/e3sconf/201910000061 ◽

2019 ◽

Vol 100 ◽

pp. 00061 ◽

Cited By ~ 1

Author(s):

Adriana Osińska ◽

Ewa Korzeniewska ◽

Monika Harnisz ◽

Sebastian Niestępski ◽

Piotr Jachimowicz

Keyword(s):

Escherichia Coli ◽

Wastewater Treatment ◽

River Water ◽

Municipal Wastewater ◽

High Efficiency ◽

Treated Wastewater ◽

Resistant Bacteria ◽

Antibiotic Resistant Bacteria ◽

Antibiotic Resistant ◽

E Coli

Wastewater treatment plants (WWTPs) are major reservoirs of antibiotic-resistant bacteria (ARB) which are transported to the natural environment with discharged effluents. Samples of untreated wastewater (UWW) and treated wastewater (TWW) from four municipal WWTPs and samples of river water collected upstream (URW) and downstream (DRW) from the effluent discharge point were analyzed in the study. The total counts of bacteria resistant to β-lactams and tetracyclines and the counts of antibiotic-resistant Escherichia coli were determined. Antibiotic-resistant bacteria, including antibiotic-resistant E. coli, were removed with up to 99.9% efficiency in the evaluated WWTPs. Despite the above, ARB counts in TWW samples were high at up to 1.25x105 CFU/mL in winter and 1.25x103 CFU/mL in summer. Antibiotic-resistant bacteria were also abundant (up to 103 CFU/ml) in URW and DRW samples collected in winter and summer. In both UWW and TWW samples, the counts of ARB and antibiotic-resistant E. coli were at least one order of magnitude lower in summer than in winter. The study revealed that despite the high efficiency of bacterial removal in the wastewater treatment processes, considerable amounts of ARB are released into the environment with TWW and that the percentage of ARB in total bacterial counts increases after wastewater treatment.

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Fate of two strains of extended-spectrum beta-lactamase producing Escherichia coli in constructed wetland microcosm sediments: survival and change in antibiotic resistance profiles

Water Science & Technology ◽

10.2166/wst.2019.153 ◽

2019 ◽

Vol 79 (8) ◽

pp. 1550-1560 ◽

Cited By ~ 1

Author(s):

Anne-Laure Vivant ◽

Catherine Boutin ◽

Stéphanie Prost-Boucle ◽

Sandrine Papias ◽

Christine Ziebal ◽

...

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Antibiotic Resistance Genes ◽

Free Water ◽

Resistant Bacteria ◽

Rapid Decline ◽

Beta Lactamase ◽

Antibiotic Resistant ◽

Extended Spectrum Beta Lactamase ◽

Extended Spectrum

Abstract Free water surface constructed wetlands (FWS CW) are efficient technologies to limit the transfer of antibiotic resistant bacteria (ARB) originating from urban effluents into the aquatic environment. However, the decrease in ARB from inflow to outflow through the FWS CW may be explained by their transfer from the water body to the sediment. To investigate the behavior of ARB in the sediment of a FWS CW, we inoculated three microcosms with two strains of extended-spectrum beta-lactamase producing Escherichia coli (ESBL E. coli) belonging to two genotypes. Microcosms were composed of two sediments collected at two locations of an FWS CW from which the strains were isolated. Phragmites were planted in one of the microcosms. The survival curves of the two strains were close regardless of the genotype and the type of sediment. After a rapid decline, both strains were able to survive at low level in the sediments for 50 days. Their fate was not affected by the presence of phragmites. Changes in the bla content and antibiotic resistance of the inoculated strains were observed after three weeks of incubation, indicating that FWS CW sediments are favorable environments for spread of antibiotic resistance genes and for the acquisition of new antibiotic resistance.

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Limited Transmission ofblaCTX-M-9-Type-Positive Escherichia coli between Humans and Poultry in Vietnam

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00517-15 ◽

2015 ◽

Vol 59 (6) ◽

pp. 3574-3577 ◽

Cited By ~ 12

Author(s):

Shuhei Ueda ◽

Bui Thi Kim Ngan ◽

Bui Thi Mai Huong ◽

Itaru Hirai ◽

Le Danh Tuyen ◽

...

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Antibiotic Resistance Genes ◽

Phylogenetic Group ◽

Content Type ◽

E Coli ◽

Plasmid Replicon ◽

Vietnamese Community ◽

Group Compositions ◽

Sequence Types

ABSTRACTWe examined whetherEscherichia coliisolates that produce CTX-M-9-type extended-spectrum β-lactamases (ESBL) are transferred between humans and chickens in a Vietnamese community. The phylogenetic group compositions, sequence types, antimicrobial resistance profiles, the prevalence of plasmid antibiotic resistance genes, and the plasmid replicon types generally differed between the human and chickenE. coliisolates. Our results suggest that transmission of theblaCTX-M-9-positiveE. colibetween humans and poultry was limited.

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Clinical Epidemiology and Molecular Analysis of Extended-Spectrum-β-Lactamase-Producing Escherichia coli in Nepal: Characteristics of Sequence Types 131 and 648

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00270-15 ◽

2015 ◽

Vol 59 (6) ◽

pp. 3424-3432 ◽

Cited By ~ 24

Author(s):

Jatan Bahadur Sherchan ◽

Kayoko Hayakawa ◽

Tohru Miyoshi-Akiyama ◽

Norio Ohmagari ◽

Teruo Kirikae ◽

...

Keyword(s):

Escherichia Coli ◽

Virulence Genes ◽

Clinical Epidemiology ◽

University Teaching ◽

Drug Resistant ◽

Content Type ◽

Acquired Drug Resistance ◽

E Coli ◽

Extended Spectrum ◽

Sequence Types

ABSTRACTRecently, CTX-M-type extended-spectrum-β-lactamase (ESBL)-producingEscherichia colistrains have emerged worldwide. In particular,E. coliwith O antigen type 25 (O25) and sequence type 131 (ST131), which is often associated with the CTX-M-15 ESBL, has been increasingly reported globally; however, epidemiology reports on ESBL-producingE. coliin Asia are limited. Patients with clinical isolates of ESBL-producingE. coliin the Tribhuvan University teaching hospital in Kathmandu, Nepal, were included in this study. Whole-genome sequencing of the isolates was conducted to analyze multilocus sequence types, phylotypes, virulence genotypes, O25b-ST131 clones, and distribution of acquired drug resistance genes. During the study period, 105 patients with ESBL-producingE. coliisolation were identified, and the majority (90%) of these isolates were CTX-M-15 positive. The most dominant ST was ST131 (n= 54; 51.4%), followed by ST648 (n= 15; 14.3%). All ST131 isolates were identified as O25b-ST131 clones, subclone H30-Rx. Three ST groups (ST131, ST648, and non-ST131/648) were compared in further analyses. ST648 isolates had a proportionally higher resistance to non-β-lactam antibiotics and featured drug-resistant genes more frequently than ST131 or non-ST131/648 isolates. ST131 possessed the most virulence genes, followed by ST648. The clinical characteristics were similar among groups. More than 38% of ESBL-producingE. coliisolates were from the outpatient clinic, and pregnant patients comprised 24% of ESBL-producingE. colicases. We revealed that the high resistance of ESBL-producingE. colito multiple classes of antibiotics in Nepal is driven mainly by CTX-M-producing ST131 and ST648. Their immense prevalence in the communities is a matter of great concern.

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Escherichia coli Antibiotic Resistance Patterns from Co-Grazing and Non-Co-Grazing Livestock and Wildlife Species from Two Farms in the Western Cape, South Africa

Antibiotics ◽

10.3390/antibiotics10060618 ◽

2021 ◽

Vol 10 (6) ◽

pp. 618

Author(s):

Michaela Sannettha van den Honert ◽

Pieter Andries Gouws ◽

Louwrens Christiaan Hoffman

Keyword(s):

Escherichia Coli ◽

South Africa ◽

Antibiotic Resistance ◽

Resistant Bacteria ◽

Antibiotic Resistant ◽

E Coli ◽

Wildlife Species ◽

Resistance Patterns ◽

Antibiotic Resistance Patterns ◽

Grazing Livestock

Although limited, studies have found conflicting results on whether co-grazing results in significant antibiotic resistance transfer between species. This type of farming system can act as a vector in the geographical spread of antibiotic-resistant bacteria in the environment. The aim of this study was to determine the antibiotic-resistant patterns between co-grazing and non-co-grazing livestock and wildlife species in South Africa. Escherichia coli was isolated from the faeces of various wildlife and livestock species from two farms in South Africa and was tested for antibiotic resistance using the Kirby–Bauer disk diffusion method against chloramphenicol, nalidixic acid, ampicillin, streptomycin, sulphafurazole, and tetracycline. A selection of some common antibiotic-resistant genes (blaCMY, aadA1, sul1, sul2, tetA, and tetB) were detected using PCR. The E. coli isolates from wildlife and livestock that co-grazed showed no significant differences in antibiotic resistance patterns. However, this was not the case for tetracycline resistance as the livestock isolates were significantly more resistant than the co-grazing wildlife isolates. The E. coli isolates from the non-co-grazing livestock and wildlife had significant differences in their antibiotic susceptibility patterns; the wildlife E. coli isolates were significantly more resistant to sulphafurazole and streptomycin than the livestock isolates, whilst those isolated from the cattle were significantly more resistant to ampicillin than the wildlife and sheep isolates. The results of this study suggest that there could be an exchange of antibiotic-resistant bacteria and genes between livestock and wildlife that co-graze.

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Antibiotic resistant and extended-spectrum β-lactamase producing faecal coliforms in wastewater treatment plant effluent

10.1101/838136 ◽

2019 ◽

Author(s):

Cian Smyth ◽

Aidan O’Flaherty ◽

Fiona Walsh ◽

Thi Thuy Do

Keyword(s):

Antibiotic Resistance ◽

Wastewater Treatment ◽

Control Point ◽

Treated Wastewater ◽

Multi Drug Resistance ◽

Resistant Bacteria ◽

Faecal Coliforms ◽

Antibiotic Resistant Bacteria ◽

Antibiotic Resistant ◽

Extended Spectrum

AbstractWastewater treatment plants (WWTPs) provide optimal conditions for the maintenance and spread of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs). In this work we describe the occurrence of antibiotic resistant faecal coliforms and their mechanisms of antibiotic resistance in the effluent of two urban WWTPs in Ireland. Effluent samples were collected from two WWTPs in Spring and Autumn of 2015 and 2016. The bacterial susceptibility patterns to 13 antibiotics were determined. The phenotypic tests were carried out to identify AmpC or extended-spectrum β-lactamase (ESBL) producers. The presence of ESBL genes were detected by PCR. Plasmids carrying ESBL genes were transformed into Escherichia coli DH5α recipient and underwent plasmid replicon typing to identify incompatibility groups. More than 90% of isolated faecal coliforms were resistant to amoxicillin and ampicillin, followed by tetracycline (up to 39.82%), ciprofloxacin (up to 31.42%) and trimethoprim (up to 37.61%). Faecal coliforms resistant to colistin and imipenem were detected in all effluent samples. Up to 53.98% of isolated faecal coliforms expressed a multi-drug resistance (MRD) phenotype. AmpC production was confirmed in 5.22% of isolates. The ESBL genes were confirmed for 11.84% of isolates (9.2% of isolates carried blaTEM, 1.4% blaSHV-12, 0.2% blaCTX-M-1 and 1% blaCTX-M-15). Plasmids extracted from 52 ESBL isolates were successfully transformed into recipient E. coli. The detected plasmid incompatibility groups included the IncF group, IncI1, IncHI1/2 and IncA/C. These results provide evidence that treated wastewater is polluted with ARB and MDR faecal coliforms and are sources of ESBL-producing, carbapenem and colistin resistant Enterobacteriaceae.ImportanceAntibiotic resistant bacteria (ARB) are an emerging environmental concern with a potential impact on human health. The results provide the evidence that treated wastewater is polluted with antibiotic resistant bacteria containing mobile resistance mechanisms of importance to clinical treatment of pathogens and multi-drug resistant (MDR) faecal coliforms. They are sources of relatively high proportions of ESBL-producing Enterobacteriaceae, and include carbapenem and colistin resistant Enterobacteriaceae. The significance of this study is the identification of the role of WWTPs as a potential control point to reduce or stop the movement of ESBL, MDR and colistin resistant bacteria into the environment from further upstream sources, such as human or animal waste.

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