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730400.9973Nanofiltration as an Efficient Tertiary Wastewater Treatment: Elimination of Total Bacteria and Antibiotic Resistance Genes from the Discharged Effluent of a Full-Scale Wastewater Treatment Plant. Wastewater reuse for agricultural irrigation still raises important public health issues regarding its safety, due to the increasing presence of emerging contaminants, such as antibiotic resistant bacteria and genes, in the treated effluents. In this paper, the potential for a commercial Desal 5 DK nanofiltration membrane to be used as a tertiary treatment in the wastewater treatment plants for a more effective elimination of these pollutants from the produced effluents was assessed on laboratory scale, using a stainless steel cross-flow cell. The obtained results showed high concentrations of total bacteria and target carbapenem and (fluoro)quinolone resistance genes (bla(KPC), bla(OXA-48), bla(NDM), bla(IMP), bla(VIM), qnrA, qnrB and qnrS) not only in the discharged, but also in the reused, effluent samples, which suggests that their use may not be entirely safe. Nevertheless, the applied nanofiltration treatment achieved removal rates superior to 98% for the total bacteria and 99.99% for all the target resistance genes present in both DNA and extracellular DNA fractions, with no significant differences for these microbiological parameters between the nanofiltered and the control tap water samples. Although additional studies are still needed to fully optimize the entire process, the use of nanofiltration membranes seems to be a promising solution to substantially increase the quality of the treated wastewater effluents.202235625274
658210.9972Effective Treatment Strategies for the Removal of Antibiotic-Resistant Bacteria, Antibiotic-Resistance Genes, and Antibiotic Residues in the Effluent From Wastewater Treatment Plants Receiving Municipal, Hospital, and Domestic Wastewater: Protocol for a Systematic Review. BACKGROUND: The widespread and unrestricted use of antibiotics has led to the emergence and spread of antibiotic-resistant bacteria (ARB), antibiotic-resistance genes (ARGs), and antibiotic residues in the environment. Conventional wastewater treatment plants (WWTPs) are not designed for effective and adequate removal of ARB, ARGs, and antibiotic residues, and therefore, they play an important role in the dissemination of antimicrobial resistance (AMR) in the natural environment. OBJECTIVE: We will conduct a systematic review to determine the most effective treatment strategies for the removal of ARB, ARGs, and antibiotic residues from the treated effluent disposed into the environment from WWTPs that receive municipal, hospital, and domestic discharge. METHODS: We will search the MEDLINE, EMBASE, Web of Science, World Health Organization Global Index Medicus, and ProQuest Environmental Science Collection databases for full-text peer-reviewed journal articles published between January 2001 and December 2020. We will select only articles published in the English language. We will include studies that measured (1) the presence, concentration, and removal rate of ARB/ARGs going from WWTP influent to effluent, (2) the presence, concentration, and types of antibiotics in the effluent, and (3) the possible selection of ARB in the effluent after undergoing treatment processes in WWTPs. At least two independent reviewers will extract data and perform risk of bias assessment. An acceptable or narrative synthesis method will be followed to synthesize the data and present descriptive characteristics of the included studies in a tabular form. The study has been approved by the Ethics Review Board at the International Centre for Diarrhoeal Disease Research, Bangladesh (protocol number: PR-20113). RESULTS: This protocol outlines our proposed methodology for conducting a systematic review. Our results will provide an update to the existing literature by searching additional databases. CONCLUSIONS: Findings from our systematic review will inform the planning of proper treatment methods that can effectively reduce the levels of ARB, ARGs, and residual antibiotics in effluent, thus lowering the risk of the environmental spread of AMR and its further transmission to humans and animals. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): PRR1-10.2196/33365.202134842550
776320.9972Antibiotic resistance genes fate and removal by a technological treatment solution for water reuse in agriculture. In order to mitigate the potential effects on the human health which are associated to the use of treated wastewater in agriculture, antibiotic resistance genes (ARGs) are required to be carefully monitored in wastewater reuse processes and their spread should be prevented by the development of efficient treatment technologies. Objective of this study was the assessment of ARGs reduction efficiencies of a novel technological treatment solution for agricultural reuse of municipal wastewaters. The proposed solution comprises an advanced biological treatment (Sequencing Batch Biofilter Granular Reactor, SBBGR), analysed both al laboratory and pilot scale, followed by sand filtration and two different disinfection final stages: ultraviolet light (UV) radiation and peracetic acid (PAA) treatments. By Polymerase Chain Reaction (PCR), the presence of 9 ARGs (ampC, mecA, ermB, sul1, sul2, tetA, tetO, tetW, vanA) were analysed and by quantitative PCR (qPCR) their removal was determined. The obtained results were compared to the reduction of total bacteria (16S rDNA gene) and of a faecal contamination indicator (Escherichia coli uidA gene). Only four of the analysed genes (ermB, sul1, sul2, tetA) were detected in raw wastewater and their abundance was estimated to be 3.4±0.7 x10(4) - 9.6±0.5 x10(9) and 1.0±0.3 x10(3) to 3.0±0.1 x10(7) gene copies/mL in raw and treated wastewaters, respectively. The results show that SBBGR technology is promising for the reduction of ARGs, achieving stable removal performance ranging from 1.0±0.4 to 2.8±0.7 log units, which is comparable to or higher than that reported for conventional activated sludge treatments. No reduction of the ARGs amount normalized to the total bacteria content (16S rDNA), was instead obtained, indicating that these genes are removed together with total bacteria and not specifically eliminated. Enhanced ARGs removal was obtained by sand filtration, while no reduction was achieved by both UV and PAA disinfection treatments tested in our study.201627450254
730530.9971Inactivation of antibiotic-resistant bacteria in hospital wastewater by ozone-based advanced water treatment processes. The emergence and spread of antimicrobial resistance (AMR) continue on a global scale. The impacts of wastewater on the environment and human health have been identified, and understanding the environmental impacts of hospital wastewater and exploring appropriate forms of treatment are major societal challenges. In the present research, we evaluated the efficacy of ozone (O(3))-based advanced wastewater treatment systems (O(3), O(3)/H(2)O(2), O(3)/UV, and O(3)/UV/H(2)O(2)) for the treatment of antimicrobials, antimicrobial-resistant bacteria (AMRB), and antimicrobial resistance genes (AMRGs) in wastewater from medical facilities. Our results indicated that the O(3)-based advanced wastewater treatment inactivated multiple antimicrobials (>99.9%) and AMRB after 10-30 min of treatment. Additionally, AMRGs were effectively removed (1.4-6.6 log(10)) during hospital wastewater treatment. The inactivation and/or removal performances of these pollutants through the O(3)/UV and O(3)/UV/H(2)O(2) treatments were significantly (P < 0.05) better than those in the O(3) and O(3)/H(2)O(2) treatments. Altered taxonomic diversity of microorganisms based on 16S rRNA gene sequencing following the O(3)-based treatment showed that advanced wastewater treatments not only removed viable bacteria but also removed genes constituting microorganisms in the wastewater. Consequently, the objective of this study was to apply advanced wastewater treatments to treat wastewater, mitigate environmental pollution, and alleviate potential threats to environmental and human health associated with AMR. Our findings will contribute to enhancing the effectiveness of advanced wastewater treatment systems through on-site application, not only in wastewater treatment plants (WWTPs) but also in medical facilities. Moreover, our results will help reduce the discharge of AMRB and AMRGs into rivers and maintain the safety of aquatic environments.202437777130
533240.9971Dynamics of antimicrobial resistance and susceptibility profile in full-scale hospital wastewater treatment plants. Drug resistance has become a matter of great concern, with many bacteria now resist multiple antibiotics. This study depicts the occurrence of antibiotic-resistant bacteria (ARB) and resistance patterns in five full-scale hospital wastewater treatment plants (WWTPs). Samples of raw influent wastewater, as well as pre- and post-disinfected effluents, were monitored for targeted ARB and resistance genes in September 2022 and February 2023. Shifts in resistance profiles of Escherichia coli, Pseudomonas aeruginosa, and Acinetobacter baumannii antimicrobial-resistant indicators in the treated effluent compared to that in the raw wastewater were also worked out. Ceftazidime (6.78 × 10(5) CFU/mL) and cefotaxime (6.14 × 10(5) CFU/mL) resistant species showed the highest concentrations followed by ciprofloxacin (6.29 × 10(4) CFU/mL), and gentamicin (4.88 × 10(4) CFU/mL), in raw influent respectively. WWTP-D employing a combination of biological treatment and coagulation/clarification for wastewater decontamination showed promising results for reducing ARB emissions from wastewater. Relationships between treated effluent quality parameters and ARB loadings showed that high BOD(5) and nitrate levels were possibly contributing to the persistence and/or selection of ARBs in WWTPs. Furthermore, antimicrobial susceptibility tests of targeted species revealed dynamic shifts in resistance profiles through treatment processes, highlighting the potential for ARB and ARGs in hospital wastewater to persist or amplify during treatment.202439007309
328150.9971Antibiotic resistance genes prevalence prediction and interpretation in beaches affected by urban wastewater discharge. BACKGROUND: The annual death toll of over 1.2 million worldwide is attributed to infections caused by resistant bacteria, driven by the significant impact of antibiotic misuse and overuse in spreading these bacteria and their associated antibiotic resistance genes (ARGs). While limited data suggest the presence of ARGs in beach environments, efficient prediction tools are needed for monitoring and detecting ARGs to ensure public health safety. This study aims to develop interpretable machine learning methods for predicting ARGs in beach waters, addressing the challenge of black-box models and enhancing our understanding of their internal mechanisms. METHODS: In this study, we systematically collected beach water samples and subsequently isolated bacteria from these samples using various differential and selective media supplemented with different antibiotics. Resistance profiles of bacteria were determined by using Kirby-Bauer disk diffusion method. Further, ARGs were enumerated by using the quantitative polymerase chain reaction (qPCR) to detect and quantify ARGs. The obtained qPCR data and hydro-meteorological were used to create an ML model with high prediction performance and we further used two explainable artificial intelligence (xAI) model-agnostic interpretation methods to describe the internal behavior of ML model. RESULTS: Using qPCR, we detected bla(CTX-M), bla(NDM), bla(CMY), bla(OXA), bla(tetX), bla(sul1), and bla(aac(6'-Ib-cr)) in the beach waters. Further, we developed ML prediction models for bla(aac(6'-Ib-cr)), bla(sul1), and bla(tetX) using the hydro-metrological and qPCR-derived data and the models demonstrated strong performance, with R2 values of 0.957, 0.997, and 0.976, respectively. CONCLUSIONS: Our findings show that environmental factors, such as water temperature, precipitation, and tide, are among the important predictors of the abundance of resistance genes at beaches.202338024281
781360.9971A framework predicting removal efficacy of antibiotic resistance genes during disinfection processes with machine learning. Disinfection has been applied widely for the removal of antibiotic resistance genes (ARGs) to curb the spread of antibiotic resistance. Quantitative polymerase chain reaction (qPCR) is the most used method to quantify the damage of DNA thus calculating the ARG degradation during disinfection but suffers the deviation due to the limitation of amplicon length. In contrast, transformation assay more accurately measures ARG deactivation based on expression of disinfected ARG in the receiving bacteria but is typically laborious and material-intensive. This work applied machine learning (ML) to develop a framework by using qPCR results as a proxy to estimate the transformation assay measurements during disinfection with chlorine (FAC), ultraviolet (UV(254)), ozone (O(3)), and hydrogen peroxide/ultraviolet (UV/H(2)O(2)) for multiple kinds of ARGs. ARG degradation rates and deactivation rates were well predicted with the optimal correlation coefficient (R(2)) of all test sets > 0.926 and > 0.871, respectively. Besides, by concatenating the ARG degradation and deactivation predictive models, ARG removal efficiency under given disinfection conditions was directly predicted as the loss of transformation activity with R(2) > 0.828. Furthermore, an online platform was built to provide users with access to the developed ML models for rapid and accurate evaluation of ARG removal efficiency.202540179779
780970.9971Mitigating Antibiotic Resistance Genes in Wastewater by Sequential Treatment with Novel Nanomaterials. Wastewater (WW) has been widely recognized as the major sink of a variety of emerging pathogens (EPs), antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs), which may disseminate and impact wider environments. Improving and maximizing WW treatment efficiency to remove these microbial hazards is fundamentally imperative. Despite a variety of physical, biological and chemical treatment technologies, the efficiency of ARG removal is still far from satisfactory. Within our recently accomplished M-ERA.NET project, novel functionalized nanomaterials, i.e., molecularly imprinted polymer (MIP) films and quaternary ammonium salt (QAS) modified kaolin microparticles, were developed and demonstrated to have significant EP removal effectiveness on both Gram-positive bacteria (GPB) and Gram-negative bacteria (GNB) from WW. As a continuation of this project, we took the further step of exploring their ARG mitigation potential. Strikingly, by applying MIP and QAS functionalized kaolin microparticles in tandem, the ARGs prevalent in wastewater treatment plants (WWTPs), e.g., blaCTXM, ermB and qnrS, can be drastically reduced by 2.7, 3.9 and 4.9 log (copies/100 mL), respectively, whereas sul1, tetO and mecA can be eliminated below their detection limits. In terms of class I integron-integrase I (intI1), a mobile genetic element (MGE) for horizontal gene transfer (HGT), 4.3 log (copies/100 mL) reduction was achieved. Overall, the novel nanomaterials exhibit outstanding performance on attenuating ARGs in WW, being superior to their control references. This finding provides additional merit to the application of developed nanomaterials for WW purification towards ARG elimination, in addition to the proven bactericidal effect.202134063382
779880.9971Effect of earthworms in reduction and fate of antibiotic resistant bacteria (ARB) and antibiotic resistant genes (ARGs) during clinical laboratory wastewater treatment by vermifiltration. In the recent decades, the role of wastewater treatment plants has been entrenched for the dissemination of antibiotic resistant bacteria into the environment. The present study explores the dynamics of earthworms-microorganisms interactions involved in the high treatment efficacy of vermifiltration technology along with reduction of antibiotic resistant bacteria (ARB). This study is the first of its kind to investigate the performance efficacy of vermifilter (VF) for clinical laboratory wastewater treatment. The results of the study showed that earthworms and VF associated microbial community had a significant effect on Biochemical Oxygen Demand (BOD) and Chemical Oxygen Demand (COD) reduction (78-85%), coliforms and pathogen removal (>99.9%) and caused a significant shift in the prevalence pattern of ARB. Molecular profiling of resistance causing genes such as ESBL (bla(SHV), bla(TEM) and bla(CTX-M)), MRSA (mec-A) and Colistin (mcr-1) confirmed the probable mechanisms behind the resistance pattern. The microbial community diversity in the influent, earthworm's coelomic fluid and gut and filter media layers associated with the VF assists in the formation of biofilm, which helps in the removal of pathogens from the wastewater. This biofilm formation further results in a paradigm shift in the resistance profile of ARB and ARG, specifically most effective against drugs, targeting cell wall and protein synthesis inhibition such as Ampicillin, Ticarcillin, Gentamicin and Chloramphenicol. These findings further validate vermifiltration technology as a sustainable and natural treatment technology for clinical laboratory wastewater, specifically for the removal of pathogens and antibiotic resistance.202133940720
776690.9970Native Microalgae-Bacteria Consortia: A Sustainable Approach for Effective Urban Wastewater Bioremediation and Disinfection. Urban wastewater is a significant by-product of human activities. Conventional urban wastewater treatment plants have limitations in their treatment, mainly concerning the low removal efficiency of conventional and emerging contaminants. Discharged wastewater also contains harmful microorganisms, posing risks to public health, especially by spreading antibiotic-resistant bacteria and genes. Therefore, this study assesses the potential of a native microalgae-bacteria system (MBS) for urban wastewater bioremediation and disinfection, targeting NH(4)(+)-N and PO(4)(3-)-P removal, coliform reduction, and antibiotic resistance gene mitigation. The MBS showed promising results, including a high specific growth rate (0.651 ± 0.155 d(-1)) and a significant average removal rate of NH(4)(+)-N and PO(4)(3-)-P (9.05 ± 1.24 mg L(-1) d(-1) and 0.79 ± 0.06 mg L(-1) d(-1), respectively). Microalgae-induced pH increase rapidly reduces coliforms (r > 0.9), including Escherichia coli, within 3 to 6 days. Notably, the prevalence of intI1 and the antibiotic resistance genes sul1 and bla(TEM) are significantly diminished, presenting the MBS as a sustainable approach for tertiary wastewater treatment to combat eutrophication and reduce waterborne disease risks and antibiotic resistance spread.202439065189
5819100.9970Application of mNGS in the Etiological Analysis of Lower Respiratory Tract Infections and the Prediction of Drug Resistance. Lower respiratory tract infections (LRTIs) have high morbidity and mortality rates. However, traditional etiological detection methods have not been able to meet the needs for the clinical diagnosis and prognosis of LRTIs. The rapid development of metagenomic next-generation sequencing (mNGS) provides new insights for the diagnosis and treatment of LRTIs; however, little is known about how to interpret the application of mNGS results in LRTIs. In this study, lower respiratory tract specimens from 46 patients with suspected LRTIs were tested simultaneously using conventional microbiological detection methods and mNGS. Receiver operating characteristic (ROC) curves were used to evaluate the performance of the logarithm of reads per kilobase per million mapped reads [lg(RPKM)], genomic coverage, and relative abundance of the organism in predicting the true-positive pathogenic bacteria. True-positive viruses were identified according to the lg(RPKM) threshold of bacteria. We also evaluated the ability to predict drug resistance genes using mNGS. Compared to that using conventional detection methods, the false-positive detection rate of pathogenic bacteria was significantly higher using mNGS. It was concluded from the ROC curves that the lg(RPKM) and genomic coverage contributed to the identification of pathogenic bacteria, with the performance of lg(RPKM) being the best (area under the curve [AUC] = 0.99). The corresponding lg(RPKM) threshold for identifying the pathogenic bacteria was -1.35. Thirty-five strains of true-positive virus were identified based on the lg(RPKM) threshold of bacteria, with the detection of human gammaherpesvirus 4 being the highest and prone to coinfection with Pseudomonas aeruginosa, Acinetobacter baumannii, and Stenotrophomonas maltophilia. Antimicrobial susceptibility tests (AST) revealed the resistance of bacteria containing drug resistance genes (detected by mNGS). However, the drug resistance genes of some multidrug-resistant bacteria were not detected. As an emerging technology, mNGS has shown many advantages for the unbiased etiological detection and the prediction of antibiotic resistance. However, a correct understanding of mNGS results is a prerequisite for its clinical application, especially for LRTIs. IMPORTANCE LRTIs are caused by hundreds of pathogens, and they have become a great threat to human health due to the limitations of traditional etiological detection methods. As an unbiased approach to detect pathogens, mNGS overcomes such etiological diagnostic challenges. However, there is no unified standard on how to use mNGS indicators (the sequencing reads, genomic coverage, and relative abundance of each organism) to distinguish between pathogens and colonizing microorganisms or contaminant microorganisms. Here, we selected the mNGS indicator with the best identification performance and established a cutoff value for the identification of pathogens in LRTIs using ROC curves. In addition, we also evaluated the accuracy of antibiotic resistance prediction using mNGS.202235171007
7791110.9970Investigation of reduction in risk from antibiotic resistance genes in laboratory wastewater by using O(3) , ultrasound, and autoclaving. Biological laboratory wastewater containing both antibiotic-resistant bacteria (ARB) and antibiotics is a potential source of antibiotic resistance genes (ARGs). Thus, we determined the efficacy of autoclaving, a common disinfection method, in eliminating 5 ARGs (sul1, sul2, tetW, tetM, amp) and the integrase-encoding gene intI1 from laboratory wastewater. Autoclaving (15 min, 121°C) inactivated all bacteria including ARB, whereas ARGs persisted in the wastewater with limited reduction even after 60 min of treatment. Ozonation (O(3) ), ultrasound (US), O(3) /US, and autoclaving followed by O(3) were investigated for their ability to reduce ARGs in laboratory wastewater. With O(3) and O(3) /US, the reduction rate ranged from 5.44 to 7.13 log for all ARGs investigated. Wastewater treatment with US alone did not reduce ARGs under the present experimental conditions (150 W, 53 kHz). Among the four treatments, autoclaving followed by O(3) treatment showed the highest reduction rates in the shortest time; however, further optimization and investigation are needed for the advanced treatment of bio-laboratory wastewater. Overall, this study provides novel insights into ARG sources and demonstrates that advanced oxidation methods can be useful to optimize laboratory wastewater treatment for ARG inactivation. PRACTITIONER POINTS: Bio-laboratory wastewater is potential reservoir of ARGs. Conventional autoclaving was not able to reduce ARGs to a low level. Autoclaving-O(3) completely eliminate all the bacteria. Autoclaving-O(3) reduced ARGs efficiently (6.12-7.86 logs removal in 60 min).202132891064
5331120.9970Performance evaluation of ozonation for removal of antibiotic-resistant Escherichia coli and Pseudomonas aeruginosa and genes from hospital wastewater. The performance of ozonation for the removal of antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs) using Escherichia coli and Pseudomonas aeruginosa carrying ARGs from hospital wastewaters was evaluated in this study. Bacterial inactivation was determined using plate count methods and real time PCR for ARG damage (Sul1, bla(tem), bla(ctx), bla(vim) and qnrS). The reduction rate of bacterial cells and ARGs was increased by different amounts of transferred ozone dose from 11 to 45 mg/L. The concentration of 10(8) cfu/ml bacteria was reduced  to an acceptable level by ozone treatment after a 5 min contact time,  Although the removal rate was much higher for concentrations of 10(6) cfu/ml and 10(4) cfu/ml bacteria. Overall, the tendency of gene reduction by ozonation from more to less was 16S rRNA > sul1 > bla(tem) > bla(ctx) > qnrS > bla(vim). Given that plasmid-borne ARGs can potentially be transferred to other bacteria even after the disinfection process, our results can provide important insights into the fate of ARGs during hospital wastewater ozonation.202134972828
8001130.9970Exploring resistomes and microbiomes in pilot-scale microalgae-bacteria wastewater treatment systems for use in low-resource settings. Antibiotic resistance genes (ARGs) released into the environment are an emerging human and environmental health concern, including ARGs spread in wastewater treatment effluents. In low-to-middle income countries (LMICs), an alternate wastewater treatment option instead of conventional systems are low-energy, high-rate algal ponds (HRAP) that use microalgae-bacteria aggregates (MABA) for waste degradation. Here we studied the robustness of ARG removal in MABA-based pilot-scale outdoor systems for 140 days of continuous operation. The HRAP system successfully removed 73 to 88 % chemical oxygen demand and up to 97.4 % ammonia, with aggregate size increasing over operating time. Fourteen ARG classes were identified in the HRAP influent, MABA, and effluent using metagenomics, with the HRAP process reducing total ARG abundances by up to 5-fold from influent to effluent. Parallel qPCR analyses showed the HRAP system significantly reduced exemplar ARGs (p < 0.05), with 1.2 to 4.9, 2.7 to 6.3, 0 to 1.5, and 1.2 to 4.8 log-removals for sul1, tetQ, bla(KPC), and intl1 genes, respectively. Sequencing of influent, effluent and MABAs samples showed associated microbial communities differed significantly, with influent communities by Enterobacteriales (clinically relevant ARGs carrying bacteria), which were less evident in MABA and effluent. In this sense, such bacteria might be excluded from MABA due to their good settling properties and the presence of antimicrobial peptides. Microalgae-bacteria treatment systems steadily reduced ARGs from wastewater during operation time, using sunlight as the energetic driver, making them ideal for use in LMIC wastewater treatment applications.202337080313
5826140.9970Rapid and accurate sepsis diagnostics via a novel probe-based multiplex real-time PCR system. Sepsis is a critical clinical emergency that requires prompt diagnosis and intervention. Its prevalence has increased due to the aging population and increased antibiotic resistance. Early identification and the use of innovative technologies are crucial for improving patient outcomes. Modern methodologies are needed to minimize the turnaround time for diagnosis and improve outcomes. Rapid diagnostic tests and multiplex PCR are effective but have limitations in identifying a range of pathogens and target genes. Our study evaluated two novel probe-based multiplex real-time PCR systems: the SEPSI ID and SEPSI DR panels. These systems can quickly identify bacterial and fungal pathogens, alongside antibiotic resistance genes. The assays cover 29 microorganisms (gram-negative bacteria, gram-positive bacteria, yeast, and mold species), alongside 23 resistance genes and four virulence factors. A streamlined workflow uses 2 µL of broth from positive blood cultures (BCs) without nucleic acid extraction and provides results in approximately 1 h. We present the results from an evaluation of 228 BCs and 22 isolates previously characterized by whole-genome sequencing. In comparison to the reference methods, the SEPSI ID panel demonstrated a sensitivity of 96.88%, a specificity of 100%, and a PPV of 100%, whereas the SEPSI DR panel showed a sensitivity of 97.8%, a PPV of 89.7%, and a specificity of 96.7%. Both panels also identified additional pathogens and resistance-related targets not detected by conventional methods. This assay shows promise for rapidly and accurately diagnosing sepsis. Future studies should validate its performance in various clinical settings to enhance sepsis management and improve patient outcomes.IMPORTANCEWe present a new diagnostic method that enables the quick and precise identification of pathogens and resistance genes from positive blood cultures, eliminating the need for nucleic acid extraction. This technique can also be used on fresh pathogen cultures. It has the potential to greatly improve treatment protocols, leading to better patient outcomes, more responsible antibiotic use, and more efficient management of healthcare resources.202541025980
5333150.9970Antibiotic resistance profile of wastewater treatment plants in Brazil reveals different patterns of resistance and multi resistant bacteria in final effluents. Wastewater treatment plants (WWTPs) are recognized as important sources of Antibiotic Resistant Bacteria (ARBs) and Antibiotic Resistant Genes (ARGs), and might play a role in the removal and dissemination of antimicrobial resistance (AMR) in the environment. Detailed information about AMR removal by the different treatment technologies commonly applied in urban WWTPs is needed. This study investigated the occurrence, removal and characterization of ARBs in WWTPs employing different technologies: WWTP-A (conventional activated sludge-CAS), WWTP-B (UASB reactor followed by biological trickling filter) and WWTP-C (modified activated sludge followed by UV disinfection-MAS/UV). Samples of raw sewage (RI) and treated effluent (TE) were collected and, through the cultivation-based method using 11 antibiotics, the antibiotic resistance profiles were characterized in a one-year period. MAS was effective in reducing ARB counts (2 to 3 log units), compared to CAS (1 log unit) and UASB/BTF (0.5 log unit). The composition of cultivable ARB differed between RI and TE samples. Escherichia was predominant in RI (56/118); whilst in TE Escherichia (31/118) was followed by Bacillus (22/118), Shigella (14/118) and Enterococcus (14/118). Most of the isolates identified (370/394) harboured at least two ARGs and in over 80 % of the isolates, 4 or more ARG (int1, blaTEM, TetA, sul1 and qnrB) were detected. A reduction in the resistance prevalence was observed in effluents after CAS and MAS processes; whilst a slight increase was observed in treated effluents from UASB/BTF and after UV disinfection stage. The multi-drug resistance (MDR) phenotype was attributed to 84.3 % of the isolates from RI (27/32) and 63.6 % from TE (21/33) samples and 52.3 % of the isolates (34/65) were resistant to carbapenems (imipenem, meropenem, ertapenem). The results indicate that treated effluents are still a source for MDR bacteria and ARGs dissemination to aquatic environments. The importance of biological sewage treatment was reinforced by the significant reductions in ARB counts observed. However, implementation of additional treatments is needed to mitigate MDR bacteria release into the environment.202336240935
7812160.9970Using the heat generated from electrically conductive concrete slabs to reduce antibiotic resistance in beef cattle manure. Proper treatment is necessary to reduce antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) in livestock manure before land application. Conventional stockpiling suffers unreliable removal efficiency, while composting can be complicated and expensive. The objective of this study was to test the feasibility of a novel heat-based technology, i.e., stockpiling manure on conductive concrete slabs, to inactivate ARB and ARGs in beef cattle manure. In this study, two independent bench-scale trials were conducted. In both trials, samples were taken from manure piles on conductive concrete slabs and regular slabs (i.e., heated and unheated piles). In the heated pile of the first trial, 25.9% and 83.5% of the pile volume met the EPA Class A and Class B biosolids standards, respectively. For the heated pile of the second trial, the two values were 43.9% and 74.2%. In both trials, nearly all forms of the total and resistant Escherichia coli and enterococci were significantly lower in the heated piles than in the unheated piles. Besides, significant reduction of ARGs in heated piles was observed in the first trial. Through this proof-of-concept study, the new technology based on conductive concrete slabs offers an alternative manure storage method to conventional stockpiling and composting with respect to reduce ARB and ARGs in manure.202133736325
6549170.9970A Review on the Prevalence and Treatment of Antibiotic Resistance Genes in Hospital Wastewater. Antibiotic resistance is a global environmental and health threat. Approximately 4.95 million deaths were associated with antibiotic resistance in 2019, including 1.27 million deaths that were directly attributable to bacterial antimicrobial resistance. Hospital wastewater is one of the key sources for the spread of clinically relevant antibiotic resistance genes (ARGs) into the environment. Understanding the current situation of ARGs in hospital wastewater is of great significance. Here, we review the prevalence of ARGs and antibiotic-resistant bacteria (ARB) in hospital wastewater and wastewater from other places and the treatment methods used. We further discuss the intersection between ARGs and COVID-19 during the pandemic. This review highlights the issues associated with the dissemination of critical ARGs from hospital wastewater into the environment. It is imperative to implement more effective processes for hospital wastewater treatment to eliminate ARGs, particularly during the current long COVID-19 period.202540278579
3188180.9970Impact of COVID-19 pandemic on profiles of antibiotic-resistant genes and bacteria in hospital wastewater. The COVID-19 pandemic has severely affected healthcare worldwide and has led to the excessive use of disinfectants and antimicrobial agents. However, the impact of excessive disinfection measures and specific medication prescriptions on the development and dissemination of bacterial drug resistance during the pandemic remains unclear. This study investigated the influence of the pandemic on the composition of antibiotics, antibiotic resistance genes (ARGs), and pathogenic communities in hospital wastewater using ultra-performance liquid chromatography-tandem mass spectrometry and metagenome sequencing. The overall level of antibiotics decreased after the COVID-19 outbreak, whereas the abundance of various ARGs increased in hospital wastewater. After COVID-19 outbreak, bla(OXA), sul2, tetX, and qnrS had higher concentrations in winter than in summer. Seasonal factors and the COVID-19 pandemic have affected the microbial structure in wastewater, especially of Klebsiella, Escherichia, Aeromonas, and Acinetobacter. Further analysis revealed the co-existence of qnrS, bla(NDM), and bla(KPC) during the pandemic. Various ARGs significantly correlated with mobile genetic elements, implying their potential mobility. A network analysis revealed that many pathogenic bacteria (Klebsiella, Escherichia, and Vibrio) were correlated with ARGs, indicating the existence of multi-drug resistant pathogens. Although the calculated resistome risk score did not change significantly, our results suggest that the COVID-19 pandemic shifted the composition of residual antibiotics and ARGs in hospital wastewater and contributed to the dissemination of bacterial drug resistance.202337399936
3284190.9970Second life of water and wastewater in the context of circular economy - Do the membrane bioreactor technology and storage reservoirs make the recycled water safe for further use? In recent years water demand drastically increased which is particularly evident in tourism-burdened mountain regions. In these areas, climate neutral circular economy strategies to minimize human impact on the environment can be successfully applied. Among these strategies, treated wastewater reuse and retaining water in storage reservoirs deserve particular attention. This study aimed to determine if recycled water produced with two circular economy systems, namely membrane bioreactor treatment plant (MBR) with UV-light effluent disinfection and a storage reservoir, is safe enough for further use in green areas irrigation in summer and artificial snow production in winter. The assessment was based on the presence and concentration of antimicrobial agents, antibiotic resistant bacteria, antibiotic resistance genes, bacterial community composition and diversity. The treated water and wastewater was compared with natural water in their vicinity. Both systems fulfill the criteria set by the European Union in terms of reclaimed water suitable for reuse. Although the MBR/UV light wastewater treatment substantially reduced the numbers of E. coli and E. faecalis (from e.g. 32,000 CFU/100 ml to 20 CFU/100 ml and 15,000 CFU/100 ml to nearly 0 CFU/ml), bacteria resistant to ampicillin, aztreonam, cefepime, ceftazidime, ertapenem and tigecycline, as well as ESBL-positive and multidrug resistant E. coli were highly prevalent in MBR-treated wastewater (88.9 %, 55.6 %, 33.3 %, 22.2 % and 11.1 % and 44.4 and 55.6 %, respectively). Applying additional tertiary treatment technology is recommended. Retaining water in storage reservoirs nearly eliminated bacterial contaminants (e.g. E. coli dropped from 350 CFU/100 ml to 10 CFU/100 ml), antibiotic resistant bacteria, resistance genes (none detected in the storage reservoir) and antibiotics (only enrofloxacin detected once in the concentration of 3.20 ng/l). Findings of this study point to the limitations of solely culture-based assessment of reclaimed water and wastewater while they may prove useful in risk management and prevention in wastewater reuse.202438378066