The Abundance of Genes Encoding ESBL, pAmpC and Non-β-Lactam Resistance in Multidrug-Resistant Enterobacteriaceae Recovered From Wastewater Effluents

The aquatic environments play a critical function in the widespread of antibiotic-resistant bacteria, ultimately impacting human health. We evaluated the abundance of Enterobacteriaceae and the various resistance gene determinants in final effluents of wastewater treatment plants (WWTPs) in the Eastern Cape Province, Republic of South Africa. A total of 44 presumptive Enterobacteriaceae was recovered following standard isolation methods. Upon molecular confirmation through Matrix-Assisted Laser Desorption/Ionization Time of flight (MALDI TOF), a high predominance of Klebsiella pneumoniae (24%) was noted. All the confirmed isolates (n = 29) subjected to a panel of eighteen antibiotics categorized into eleven different classes were multidrug-resistant (MDR). They displayed resistance against antibiotics in more than three different classes. Notably, one E. cloacae exhibited resistance against all the antibiotics assayed. The multiple antibiotic resistance indices (MARI) ranged from 0.22 to 1.0, indicating an environment with high pressure of antibiotics. Conventional Polymerase Chain Reaction (PCR) analysis showed that 72.4% of the isolates harboured at least one β-lactamase genetic determinant. The most predominant extended-spectrum β-lactamases were blaCTX-M-group (21.5%), blaTEM (20%), and blaSHV (16.9%) while the most predominant plasmid-mediated AmpC-type β-lactamases were blaCIT and blaACC (25.9%) and blaEBC (22.2%). A total of 86.2% of the Enterobacteriaceae harboured a minimum of one non-β-lactam resistance gene determinant with predominance observed in catII (25%), sulII (15.8%), and sulI (14.5%). These results demonstrate an abundance of multidrug-resistant (MDR) Enterobacteriaceae in WWTPs’ effluents. The study confirms the need to optimize current WWTPs’ processes to improve the quality of effluents and ultimately reduce the potential risks of using such water when discharged into the environment.

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

Microorganisms ◽

10.3390/microorganisms8091425 ◽

2020 ◽

Vol 8 (9) ◽

pp. 1425

Author(s):

Lara Pérez-Etayo ◽

David González ◽

José Leiva ◽

Ana Isabel Vitas

Keyword(s):

Wastewater Treatment Plants ◽

Multidrug Resistant ◽

World Health ◽

Resistant Bacteria ◽

Aquatic Environments ◽

Multidrug Resistant Bacteria ◽

Antibiotic Resistant ◽

The North ◽

New Antibiotics ◽

Health Organization

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.

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Microbiota restoration reduces antibiotic-resistant bacteria gut colonization in patients with recurrent Clostridioides difficile infection from the open-label PUNCH CD study

Genome Medicine ◽

10.1186/s13073-021-00843-9 ◽

2021 ◽

Vol 13 (1) ◽

Cited By ~ 1

Author(s):

Amy Langdon ◽

◽

Drew J. Schwartz ◽

Christopher Bulow ◽

Xiaoqing Sun ◽

...

Keyword(s):

Antibiotic Resistance ◽

Gut Microbiota ◽

Resistance Gene ◽

Resistance Genes ◽

Multidrug Resistant ◽

Resistant Bacteria ◽

Antibiotic Resistant Bacteria ◽

Open Label ◽

Antibiotic Resistant ◽

Clostridioides Difficile

Abstract Background Once antibiotic-resistant bacteria become established within the gut microbiota, they can cause infections in the host and be transmitted to other people and the environment. Currently, there are no effective modalities for decreasing or preventing colonization by antibiotic-resistant bacteria. Intestinal microbiota restoration can prevent Clostridioides difficile infection (CDI) recurrences. Another potential application of microbiota restoration is suppression of non-C. difficile multidrug-resistant bacteria and overall decrease in the abundance of antibiotic resistance genes (the resistome) within the gut microbiota. This study characterizes the effects of RBX2660, a microbiota-based investigational therapeutic, on the composition and abundance of the gut microbiota and resistome, as well as multidrug-resistant organism carriage, after delivery to patients suffering from recurrent CDI. Methods An open-label, multi-center clinical trial in 11 centers in the USA for the safety and efficacy of RBX2660 on recurrent CDI was conducted. Fecal specimens from 29 of these subjects with recurrent CDI who received either one (N = 16) or two doses of RBX2660 (N = 13) were analyzed secondarily. Stool samples were collected prior to and at intervals up to 6 months post-therapy and analyzed in three ways: (1) 16S rRNA gene sequencing for microbiota taxonomic composition, (2) whole metagenome shotgun sequencing for functional pathways and antibiotic resistome content, and (3) selective and differential bacterial culturing followed by isolate genome sequencing to longitudinally track multidrug-resistant organisms. Results Successful prevention of CDI recurrence with RBX2660 correlated with taxonomic convergence of patient microbiota to the donor microbiota as measured by weighted UniFrac distance. RBX2660 dramatically reduced the abundance of antibiotic-resistant Enterobacteriaceae in the 2 months after administration. Fecal antibiotic resistance gene carriage decreased in direct relationship to the degree to which donor microbiota engrafted. Conclusions Microbiota-based therapeutics reduce resistance gene abundance and resistant organisms in the recipient gut microbiome. This approach could potentially reduce the risk of infections caused by resistant organisms within the patient and the transfer of resistance genes or pathogens to others. Trial registration ClinicalTrials.gov, NCT01925417; registered on August 19, 2013.

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Multidrug-Resistant Salmonella Isolates from Swine in the Eastern Cape Province, South Africa

Journal of Food Protection ◽

10.4315/0362-028x.jfp-15-224 ◽

2016 ◽

Vol 79 (7) ◽

pp. 1234-1239 ◽

Cited By ~ 16

Author(s):

CHINWE JULIANA IWU ◽

BENSON CHUKS IWERIEBOR ◽

LARRY CHIKWELU OBI ◽

ALBERTUS KOTZE BASSON ◽

ANTHONY IFEANYI OKOH

Keyword(s):

South Africa ◽

Growth Promotion ◽

Resistance Index ◽

Multidrug Resistant ◽

Farm Animals ◽

Diffusion Method ◽

Resistant Bacteria ◽

Eastern Cape ◽

Disk Diffusion Method ◽

Genes Encoding

ABSTRACT The exposure of farm animals to antimicrobials for treatment, prophylaxis, or growth promotion can select for resistant bacteria that can be transmitted to humans, and Salmonella as an important zoonotic pathogen can act as a potential reservoir of antimicrobial resistance determinants. We assessed the antibiogram profiles of Salmonella species isolated from pig herds in two commercial farms in South Africa. Two hundred fifty-eight presumptive Salmonella isolates were recovered from the fecal samples of 500 adult pigs. Specific primers targeting Salmonella serogroups A, B, C1, C2, and D were used to determine the prevalence of different serogroups. Only serogroup A (n =48) was detected, while others were not. Antimicrobial susceptibility of the confirmed Salmonella serogroup A isolates was performed by using the disk diffusion method against a panel of 18 antibiotics. All the 48 isolates were resistant to tetracycline and oxytetracycline, while 75% were resistant to ampicillin, sulphamethoxazole-trimethoprim, nalidixic acid, and streptomycin. All the isolates exhibited multidrug resistance, with the predominant phenotype being against 11 antibiotics, and multiple antibiotic resistance index ranged between 0.3 and 0.6. The incidence of genes encoding resistance against ampicillin (ampC), tetracycline (tetA), and streptomycin (strA) were 54, 61, and 44%, respectively. We conclude that healthy pigs are potential reservoirs of multidrug-resistant Salmonella that could be transmitted to humans through the food chain and, hence, a significant public health threat.

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Hospital Wastewater—Important Source of Multidrug Resistant Coliform Bacteria with ESBL-Production

International Journal of Environmental Research and Public Health ◽

10.3390/ijerph17217827 ◽

2020 ◽

Vol 17 (21) ◽

pp. 7827

Author(s):

Kristína Lépesová ◽

Petra Olejníková ◽

Tomáš Mackuľak ◽

Klára Cverenkárová ◽

Monika Krahulcová ◽

...

Keyword(s):

Efflux Pump ◽

Wastewater Treatment Plants ◽

Resistance Mechanism ◽

Multidrug Resistant ◽

Coliform Bacteria ◽

Resistant Bacteria ◽

Hospital Wastewater ◽

Multidrug Resistant Bacteria ◽

Antibiotic Resistant ◽

Extended Spectrum

This work compares the prevalence of antibiotic resistant coliform bacteria in hospital wastewater effluents in Slovak (SR) and Czech Republic (ČR). It also describes selected antibiotic resistant isolates in view of resistance mechanism and virulence factor. The highest number of multidrug resistant bacteria was detected in samples from the hospital in Valašské Meziříčí (ČR). More than half of resistant isolates showed multidrug resistance phenotype as well as strong ability to form biofilm. In 42% of isolates efflux pump overproduction was detected together with tetA and tetE genes. The production of extended-spectrum β-lactamases in coliform isolates was encoded mainly by blaTEM, blaCTX-M-2 and blaCTX-M-8/25 genes. About 62% of resistants contained a combination of two or more extended spectrum beta-lactamases (ESBL) genes. Our results strengthen the fact that hospital effluents are a source of multidrug resistant bacteria which can spread their resistance genes to other bacteria in wastewater treatment plants (WWTPs). Accordingly, hospital wastewater should be better treated before it enters urban sewerage.

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Multiresistant Bacteria Isolated from Intestinal Faeces of Farm Animals in Austria

Antibiotics ◽

10.3390/antibiotics10040466 ◽

2021 ◽

Vol 10 (4) ◽

pp. 466

Author(s):

Herbert Galler ◽

Josefa Luxner ◽

Christian Petternel ◽

Franz F. Reinthaler ◽

Juliana Habib ◽

...

Keyword(s):

Meat Products ◽

Animal Husbandry ◽

Multidrug Resistant ◽

Farm Animals ◽

Resistant Bacteria ◽

Antibiotic Resistant Bacteria ◽

Antibiotic Resistant ◽

E Coli ◽

Multiresistant Bacteria ◽

Vancomycin Resistant

In recent years, antibiotic-resistant bacteria with an impact on human health, such as extended spectrum β-lactamase (ESBL)-containing Enterobacteriaceae, methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci (VRE), have become more common in food. This is due to the use of antibiotics in animal husbandry, which leads to the promotion of antibiotic resistance and thus also makes food a source of such resistant bacteria. Most studies dealing with this issue usually focus on the animals or processed food products to examine the antibiotic resistant bacteria. This study investigated the intestine as another main habitat besides the skin for multiresistant bacteria. For this purpose, faeces samples were taken directly from the intestines of swine (n = 71) and broiler (n = 100) during the slaughter process and analysed. All samples were from animals fed in Austria and slaughtered in Austrian slaughterhouses for food production. The samples were examined for the presence of ESBL-producing Enterobacteriaceae, MRSA, MRCoNS and VRE. The resistance genes of the isolated bacteria were detected and sequenced by PCR. Phenotypic ESBL-producing Escherichia coli could be isolated in 10% of broiler casings (10 out of 100) and 43.6% of swine casings (31 out of 71). In line with previous studies, the results of this study showed that CTX-M-1 was the dominant ESBL produced by E. coli from swine (n = 25, 83.3%) and SHV-12 from broilers (n = 13, 81.3%). Overall, the frequency of positive samples with multidrug-resistant bacteria was lower than in most comparable studies focusing on meat products.

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Antibiotic-Resistant Bacteria in Clams—A Study on Mussels in the River Rhine

Antibiotics ◽

10.3390/antibiotics10050571 ◽

2021 ◽

Vol 10 (5) ◽

pp. 571

Author(s):

Nicole Zacharias ◽

Iris Löckener ◽

Sarah M. Essert ◽

Esther Sib ◽

Gabriele Bierbaum ◽

...

Keyword(s):

Bacterial Infections ◽

Sewage Treatment ◽

Sewage Treatment Plant ◽

Treatment Plant ◽

Multidrug Resistant ◽

Resistant Bacteria ◽

River Rhine ◽

Antibiotic Resistant Bacteria ◽

Surrounding Water ◽

Antibiotic Resistant

Bacterial infections have been treated effectively by antibiotics since the discovery of penicillin in 1928. A worldwide increase in the use of antibiotics led to the emergence of antibiotic resistant strains in almost all bacterial pathogens, which complicates the treatment of infectious diseases. Antibiotic-resistant bacteria play an important role in increasing the risk associated with the usage of surface waters (e.g., irrigation, recreation) and the spread of the resistance genes. Many studies show that important pathogenic antibiotic-resistant bacteria can enter the environment by the discharge of sewage treatment plants and combined sewage overflow events. Mussels have successfully been used as bio-indicators of heavy metals, chemicals and parasites; they may also be efficient bio-indicators for viruses and bacteria. In this study an influence of the discharge of a sewage treatment plant could be shown in regard to the presence of E. coli in higher concentrations in the mussels downstream the treatment plant. Antibiotic-resistant bacteria, resistant against one or two classes of antibiotics and relevance for human health could be detected in the mussels at different sampling sites of the river Rhine. No multidrug-resistant bacteria could be isolated from the mussels, although they were found in samples of the surrounding water body.

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Testudines as Sentinels for Monitoring the Dissemination of Antibiotic Resistance in Marine Environments: An Integrative Review

Antibiotics ◽

10.3390/antibiotics10070775 ◽

2021 ◽

Vol 10 (7) ◽

pp. 775

Author(s):

Kezia Drane ◽

Roger Huerlimann ◽

Michelle Power ◽

Anna Whelan ◽

Ellen Ariel ◽

...

Keyword(s):

Antibiotic Resistance ◽

Anthropogenic Activities ◽

Sea Turtles ◽

Multidrug Resistant ◽

Resistant Bacteria ◽

Marine Environments ◽

Beta Lactam ◽

Antibiotic Resistant ◽

Sensitivity Tests ◽

Standard Culture

Dissemination of antibiotic resistance (AR) in marine environments is a global concern with a propensity to affect public health and many ecosystems worldwide. We evaluated the use of sea turtles as sentinel species for monitoring AR in marine environments. In this field, antibiotic-resistant bacteria have been commonly identified by using standard culture and sensitivity tests, leading to an overrepresentation of specific, culturable bacterial classes in the available literature. AR was detected against all major antibiotic classes, but the highest cumulative global frequency of resistance in all represented geographical sites was against the beta-lactam class by a two-fold difference compared to all other antibiotics. Wastewater facilities and turtle rehabilitation centres were associated with higher incidences of multidrug-resistant bacteria (MDRB) accounting for an average of 58% and 49% of resistant isolates, respectively. Furthermore, a relatively similar prevalence of MDRB was seen in all studied locations. These data suggest that anthropogenically driven selection pressures for the development of AR in sea turtles and marine environments are relatively similar worldwide. There is a need, however, to establish direct demonstrable associations between AR in sea turtles in their respective marine environments with wastewater facilities and other anthropogenic activities worldwide.

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On-Site Pilot Testing of Hospital Wastewater Ozonation to Reduce Pharmaceutical Residues and Antibiotic-Resistant Bacteria

Antibiotics ◽

10.3390/antibiotics10060684 ◽

2021 ◽

Vol 10 (6) ◽

pp. 684

Author(s):

Sofia Svebrant ◽

Robert Spörndly ◽

Richard H. Lindberg ◽

Therese Olsen Sköldstam ◽

Jim Larsson ◽

...

Keyword(s):

Municipal Wastewater ◽

Wastewater Treatment Plants ◽

High Flow ◽

Resistant Bacteria ◽

Municipal Wastewater Treatment ◽

Antibiotic Resistant Bacteria ◽

Antibiotic Resistant ◽

Untreated Wastewater ◽

Significant Difference ◽

Hospital Sewage

Hospital sewage constitutes an important point source for antibiotics and antibiotic-resistant bacteria due to the high antibiotic use. Antibiotic resistance can develop and cause problems in sewage systems within hospitals and municipal wastewater treatment plants, thus, interventions to treat hospital sewage on-site are important. Ozonation has proven effective in treating relatively clean wastewater, but the effect on untreated wastewater is unclear. Therefore, we piloted implementation of ozonation to treat wastewater in a tertiary hospital in Uppsala, Sweden. We measured active pharmaceutical ingredients (APIs) using liquid chromatography-mass spectrometry and antibiotic-resistant Enterobacteriaceae using selective culturing pre- and post-ozonation. Comparing low (1 m3/h) and high (2 m3/h) flow, we obtained a ‘dose-dependent’ effect of API reduction (significant reduction of 12/29 APIs using low and 2/29 APIs using high flow, and a mean reduction of antibiotics of 41% using low vs. 6% using high flow, 25% vs. 6% for all APIs). There was no significant difference in the amount of antibiotic-resistant Enterobacteiaceae pre- and post-ozonation. Our results demonstrate that ozonation of untreated wastewater can reduce API content. However, due to the moderate API decrease and numerous practical challenges in the on-site setting, this specific ozonation system is not suitable to implement at full scale in our hospital.

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Prevalence of Antibiotic-Resistant Bacteria ESKAPE among Healthy People Estimated by Monitoring of Municipal Wastewater

Antibiotics ◽

10.3390/antibiotics10050495 ◽

2021 ◽

Vol 10 (5) ◽

pp. 495

Author(s):

Masateru Nishiyama ◽

Susan Praise ◽

Keiichi Tsurumaki ◽

Hiroaki Baba ◽

Hajime Kanamori ◽

...

Keyword(s):

Antibiotic Resistance ◽

General Population ◽

Municipal Wastewater ◽

Treatment Plant ◽

Multidrug Resistant ◽

Resistant Bacteria ◽

Municipal Wastewater Treatment ◽

Antibiotic Resistant ◽

Carbapenem Resistant ◽

Wastewater Samples

There is increasing attention toward factors that potentially contribute to antibiotic resistance (AR), as well as an interest in exploring the emergence and occurrence of antibiotic resistance bacteria (ARB). We monitored six ARBs that cause hospital outbreaks in wastewater influent to highlight the presence of these ARBs in the general population. We analyzed wastewater samples from a municipal wastewater treatment plant (MWWTP) and hospital wastewater (HW) for six species of ARB: Carbapenem-resistant Enterobacteria (CARBA), extended-spectrum β-lactamase producing Enterobacteria (ESBL), multidrug-resistant Acinetobacter (MDRA), multidrug-resistant Pseudomonas aeruginosa (MDRP), methicillin-resistant Staphylococcus aureus (MRSA), and vancomycin-resistant Enterococci (VRE). We registered a high percentage of ARBs in MWWTP samples (>66%) for all ARBs except for MDRP, indicating a high prevalence in the population. Percentages in HW samples were low (<78%), and no VRE was detected throughout the study. CARBA and ESBL were detected in all wastewater samples, whereas MDRA and MRSA had a high abundance. This result demonstrated the functionality of using raw wastewater at MWWTP to monitor the presence and extent of ARB in healthy populations. This kind of surveillance will contribute to strengthening the efforts toward reducing ARBs through the detection of ARBs to which the general population is exposed.

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Time-Resolved Transcriptional Profiling of Epithelial Cells Infected by Intracellular Acinetobacter baumannii

Microorganisms ◽

10.3390/microorganisms9020354 ◽

2021 ◽

Vol 9 (2) ◽

pp. 354

Author(s):

Nuria Crua Asensio ◽

Javier Macho Rendón ◽

Marc Torrent Burgas

Keyword(s):

Acinetobacter Baumannii ◽

Transcriptional Profiling ◽

Multidrug Resistant ◽

Host Cells ◽

Resistant Bacteria ◽

Multidrug Resistant Bacteria ◽

Time Resolved ◽

Antibiotic Resistant ◽

Common Features ◽

Profile Changes

The rise in the number of antibiotic-resistant bacteria has become a serious threat to health, making it important to identify, characterize and optimize new molecules to help us to overcome the infections they cause. It is well known that Acinetobacter baumannii has a significant capacity to evade the actions of antibacterial drugs, leading to its emergence as one of the bacteria responsible for hospital and community-acquired infections. Nonetheless, how this pathogen infects and survives inside the host cell is unclear. In this study, we analyze the time-resolved transcriptional profile changes observed in human epithelial HeLa cells after infection by A. baumannii, demonstrating how it survives in host cells and starts to replicate 4 h post infection. These findings were achieved by sequencing RNA to obtain a set of Differentially Expressed Genes (DEGs) to understand how bacteria alter the host cells’ environment for their own benefit. We also determine common features observed in this set of genes and identify the protein–protein networks that reveal highly-interacted proteins. The combination of these findings paves the way for the discovery of new antimicrobial candidates for the treatment of multidrug-resistant bacteria.

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