Antibiotic resistances from slaughterhouse effluents and enhanced antimicrobial blue light technology for wastewater decontamionation. - Related Documents




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780301.0000Antibiotic resistances from slaughterhouse effluents and enhanced antimicrobial blue light technology for wastewater decontamionation. The frequencies of 6 different facultative pathogenic bacteria of the ESKAPE group (priority list WHO) and a total of 14 antibiotic resistance genes (ARGs) with different priorities for human medicine were quantified in wastewaters of poultry and pig slaughterhouses using molecular biological approaches. Raw sewage from poultry and pig slaughterhouses was found to be contaminated not only with facultative pathogenic bacteria but also with various categories of clinically relevant ARGs, including ARGs against the reserve antibiotics group. The concentration of the different gene targets decreased after on-site conventional biological or advanced oxidative wastewater treatments, but was not eliminated. Hence, the antimicrobial BlueLight (aBL) in combination with a porphyrin photo-sensitizer was studied with ESKAPE bacteria and real slaughterhouse wastewaters. The applied broad LED-based blue light (420-480 nm) resulted in groups of sensitive, intermediate, and non-sensitive ESKAPE bacteria. The killing effect of aBL was increased in the non-sensitive bacteria Klebsiella pneumoniae and Enterococcus faecium due to the addition of porphyrins in concentrations of 10(-6) M. Diluted slaughterhouse raw wastewater was treated with broad spectrum aBL and in combination with porphyrin. Here, the presence of the photo-sensitizer enhanced the aBL biocidal impact.202337924165
342110.9996Survival of antibiotic resistant bacteria following artificial solar radiation of secondary wastewater effluent. Urban wastewater treatment plant effluents represent one of the major emission sources of antibiotic-resistant bacteria (ARB) in natural aquatic environments. In this study, the effect of artificial solar radiation on total culturable heterotrophic bacteria and ARB (including amoxicillin-resistant, ciprofloxacin-resistant, rifampicin-resistant, sulfamethoxazole-resistant, and tetracycline-resistant bacteria) present in secondary effluent was investigated. Artificial solar radiation was effective in inactivating the majority of environmental bacteria, however, the proportion of strains with ciprofloxacin-resistance and rifampicin-resistance increased in the surviving populations. Isolates of Pseudomonas putida, Serratia marcescens, and Stenotrophomonas maltophilia nosocomial pathogens were identified as resistant to solar radiation and to at least three antibiotics. Draft genome sequencing and typing revealed isolates carrying multiple resistance genes; where S. maltophilia (resistant to all studied antibiotics) sequence type was similar to strains isolated in blood infections. Results from this study confirm that solar radiation reduces total bacterial load in secondary effluent, but may indirectly increase the relative abundance of ARB.201829898509
342720.9996Annual changes in the occurrence of antibiotic-resistant coliform bacteria and enterococci in municipal wastewater. Wastewater contains subinhibitory concentrations of different micropollutants such as antibiotics that create selective pressure on bacteria. This phenomenon is also caused by insufficient wastewater treatment technology leading to the development and spread of antibiotic-resistant bacteria and resistance genes into the environment. Therefore, this work focused on monitoring of antibiotic-resistant coliform bacteria and enterococci in influent and effluent wastewaters taken from the second biggest wastewater treatment plant (Petržalka) in the capital of Slovakia during 1 year. Antibiotic-resistant strains were isolated, identified, and characterized in terms of susceptibility and biofilm production. All of 27 antibiotic-resistant isolates were identified mainly as Morganella morganii, Citrobacter spp., and E. coli. Multidrug-resistance was detected in 58% of isolated strains. All tested isolates could form biofilm; two strains were very strong producers, and 74% formed biofilm by strong intensity. The flow rate of the influent wastewater had a more significant impact on the number of studied bacteria than the temperature. Graphical abstract.201931049859
337830.9996Antibiotic-resistant bacteria in hospital wastewater treatment plant effluent and the possible consequences of its reuse in agricultural irrigation. Wastewater from hospitals should be monitored precisely and treated properly before discharge and reuse to avoid epidemic and pandemic complications, as it contains hazardous pollutants for the ecosystem. Antibiotic residues in treated hospital wastewater effluents constitute a major environmental concern since they resist various wastewater treatment processes. The emergence and spread of multi-drug-resistant bacteria, that cause public health problems, are therefore always a major concern. The aims and objectives of this study were mainly to characterize the chemical and microbial properties of the hospital effluent of wastewater treatment plant (WWTP) before discharge to the environment. Special attention was paid to the presence of multiple resistant bacteria and the effects of hospital effluent reuse in irrigation on zucchini as an economically important plant. The risk of cell-free DNA carrying antibiotic resistance genes contained in the hospital effluent as a long-lasting hazard had been discussed. In this study, 21 bacterial strains were isolated from the effluent of a hospital WWTP. Isolated bacteria were evaluated for multi-drug resistance ability against 5 antibiotics (Tetracycline, Ampicillin, Amoxicillin, Chloramphenicol, and Erythromycin) at a concentration of 25 ppm. Out of them, three isolates (AH-03, AH-07, and AH-13) were selected because they recorded the highest growth in presence of tested antibiotics. Selected isolates were identified using 16S rRNA gene sequence homology as Staphylococcus haemolyticus (AH-03), Enterococcus faecalis (AH-07), and Escherichia coli (AH-13). Their susceptibility to ascending concentrations of tested antibiotics indicated that they were all susceptible at a concentration above 50 ppm. Results of the greenhouse experiment regarding the effect of hospital WWTP effluent reuse on zucchini plant fresh weights compared to that irrigated with fresh water indicated that the former recorded a limited increase in total fresh weights (6.2 g and 5.3 g/plant, respectively). Our results demonstrated the low impact of the reuse of Hospital WWTP effluent in agriculture irrigation compared to its greater risk in transferring multiple antibiotic bacteria and antibiotic resistance genes to soil bacteria through natural transformation.202337143530
342640.9996Detection of antibiotic-resistant bacteria and their resistance genes in wastewater, surface water, and drinking water biofilms. Abstract In view of the increasing interest in the possible role played by hospital and municipal wastewater systems in the selection of antibiotic-resistant bacteria, biofilms were investigated using enterococci, staphylococci, Enterobacteriaceae, and heterotrophic bacteria as indicator organisms. In addition to wastewater, biofilms were also investigated in drinking water from river bank filtrate to estimate the occurrence of resistant bacteria and their resistance genes, thus indicating possible transfer from wastewater and surface water to the drinking water distribution network. Vancomycin-resistant enterococci were characterized by antibiograms, and the vanA resistance gene was detected by molecular biology methods, including PCR. The vanA gene was found not only in wastewater biofilms but also in drinking water biofilms in the absence of enterococci, indicating possible gene transfer to autochthonous drinking water bacteria. The mecA gene encoding methicillin resistance in staphylococci was detected in hospital wastewater biofilms but not in any other compartment. Enterobacterial ampC resistance genes encoding beta-lactamase activities were amplified by PCR from wastewater, surface water and drinking water biofilms.200319719664
342350.9996bla(TEM) and vanA as indicator genes of antibiotic resistance contamination in a hospital-urban wastewater treatment plant system. Four indicator genes were monitored by quantitative PCR in hospital effluent (HE) and in the raw and treated wastewater of the municipal wastewater treatment plant receiving the hospital discharge. The indicator genes were the class 1 integrase gene intI1, to assess the capacity of bacteria to be involved in horizontal gene transfer processes; bla(TEM), one of the most widespread antibiotic resistance genes in the environment, associated with Enterobacteriaceae; vanA, an antibiotic resistance gene uncommon in the environment and frequent in clinical isolates; and marA, part of a locus related to the stress response in Enterobacteriaceae. Variation in the abundance of these genes was analysed as a function of the type of water, and possible correlations with cultivable bacteria, antimicrobial residue concentrations, and bacterial community composition and structure were analysed. HE was confirmed as an important source of bla(TEM) and vanA genes, and wastewater treatment showed a limited capacity to remove these resistance genes. The genes bla(TEM) and vanA presented the strongest correlations with culturable bacteria, antimicrobial residues and some bacterial populations, representing interesting candidates as indicator genes to monitor resistance in environmental samples. The intI1 gene was the most abundant in all samples, demonstrating that wastewater bacterial populations hold a high potential for gene acquisition.201427873693
780260.9995Inactivating facultative pathogen bacteria and antibiotic resistance genes in wastewater using blue light irradiation combined with a photosensitizer and hydrogen peroxide. The effectiveness of antimicrobial blue light (aBL) irradiation in eliminating ten clinically significant antibiotic resistance genes (ARGs) and four taxonomic marker genes of the WHO-priority ESKAPE bacteria group from wastewater treatment plant (WWTP) effluent was examined. Experiments were conducted using an LED-driven continuous-flow photoreactor operating at wavelengths of 405 nm, 420 nm, and 460 nm. Irradiation with aBL alone was insufficient for effectively inactivating or eliminating ESKAPE bacteria and clinically relevant ARGs. The addition of the porphyrin-based photosensitizer TMPyP (10(-6) M) or the oxidative agent H₂O₂ (1 mM) resulted in several log(10) unit reductions of facultative pathogenic bacteria (FPB), their taxonomic gene markers, and target ARGs. However, the additional effects of TMPyP and H(2)O(2) were only noticeable in conjunction with aBL irradiation, as they were ineffective without it. The reduction of the different FPB and ARGs in WWTP effluents was analyzed using culturing and qPCR together with living/dead discrimination. Different FPB and ARGs showed varying susceptibility to aBL-mediated irradiation. Among the FPB, enterococci were the most sensitive, while among the ARGs bacteria carrying ermB, tetM, sul1, and bla(VIM) genes exhibited the strongest removal. This sensitivity may be due to the gene-carrying microorganism's response to aBL irradiation combined with TMPyP or H(2)O(2). Additionally, molecular biology results revealed that aBL irradiation induced up to 13 lesions per 10 kb DNA, which is hypothesized to contribute to the acute inactivation effect and prevent regrowth by inhibiting DNA repair activities.202540138903
328470.9995Second 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
529380.9995Tetracycline-Resistant Bacteria Selected from Water and Zebrafish after Antibiotic Exposure. The emergence of antibiotic-resistant pathogens due to worldwide antibiotic use is raising concern in several settings, including aquaculture. In this work, the selection of antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs) was evaluated after exposure of zebrafish to oxytetracycline (OTC) for two months, followed by a recovery period. The selection of ARB in water and fish was determined using selective media. The abundance of tetA genes was estimated through qPCR. Higher prevalence of ARB was measured in all samples exposed to the antibiotic when compared to control samples, although statistical significance was only achieved five days after exposure. Isolates recovered from samples exposed to the antibiotic were affiliated with Pseudomonas and Stenotrophomonas. Various antibiotic susceptibility profiles were detected and 37% of the isolates displayed multidrug resistance (MDR). The selection of the tetA gene was confirmed by qPCR at the highest OTC concentration tested. Two MDR isolates, tested using zebrafish embryos, caused significant mortality, indicating a potential impact on fish health and survival. Overall, our work highlights the potential impact of antibiotic contamination in the selection of potential pathogenic ARB and ARGS.202133804606
346890.9995Facultative pathogenic bacteria and antibiotic resistance genes in swine livestock manure and clinical wastewater: A molecular biology comparison. Manure contains vast amounts of biological contaminants of veterinary origin. Only few studies analyse clinically critical resistance genes against reserve antibiotics in manure. In general, resistances against these high priority antibiotics involve a high potential health risk. Therefore, their spread in the soil as well as the aquatic environment has to be prevented. Manures of 29 different swine livestock were analysed. Abundances of facultative pathogenic bacteria including representatives of the clinically critical ESKAPE-pathogens (P. aeruginosa, K. pneumoniae, A. baumannii, E. faecium) and E. coli were investigated via qPCR. Antibiotic resistance genes against commonly used veterinary antibiotics (ermB, tetM, sul1) as well as various resistance genes against important (mecA, vanA) and reserve antibiotics (bla(NDM), bla(KPC3), mcr-1), which are identified by the WHO, were also obtained by qPCR analysis. The manures of all swine livestock contained facultative pathogenic bacteria and commonly known resistance genes against antibiotics used in veterinary therapies, but more important also a significant amount of clinically critical resistance genes against reserve antibiotics for human medicine. To illustrate the impact the occurrence of these clinically critical resistance genes, comparative measurements were taken of the total wastewater of a large tertiary care hospital (n = 8). Both manure as well as raw hospital wastewaters were contaminated with significant abundances of gene markers for facultative pathogens and with critical resistance genes of reserve antibiotics associated with genetic mobile elements for horizontal gene transfer. Hence, both compartments bear an exceptional potential risk for the dissemination of facultative pathogens and critical antibiotic resistance genes.202236089145
4570100.9995Detection of sulfonamide resistance genes via in situ PCR-FISH. Due to the rising use of antibiotics and as a consequence of their concentration in the environment an increasing number of antibiotic resistant bacteria is observed. The phenomenon has a hazardous impact on human and animal life. Sulfamethoxazole is one of the sulfonamides commonly detected in surface waters and soil. The aim of the study was to detect sulfamethoxazole resistance genes in activated sludge biocenosis by use of in situ PCR and/or hybridization. So far no FISH probes for the detection of SMX resistance genes have been described in the literature. We have tested common PCR primers used for SMX resistance genes detection as FISH probes as well as a combination of in situ PCR and FISH. Despite the presence of SMX resistance genes in activated sludge confirmed via traditional PCR, the detection of the genes via microscopic visualization failed.201425115110
3430110.9995Benzalkonium chloride alters phenotypic and genotypic antibiotic resistance profiles in a source water used for drinking water treatment. Antibiotic resistance is a major public health concern. Triclosan is an antimicrobial compound with direct links to antibiotic resistance that was widely used in soaps in the U.S. until its ban by the U.S. Food and Drug Administration. Benzalkonium chloride (BAC), a quaternary ammonium compound, has widely replaced triclosan in soaps marketed as an antibacterial. BAC has been detected in surface waters and its presence will likely increase following increased use in soap products. The objective of this study was to determine the effect of BAC on relative abundance of antibiotic resistance in a bacterial community from a surface water used as a source for drinking water treatment. Bench-scale microcosm experiments were conducted with microbial communities amended with BAC at concentrations ranging from 0.1 μg L(-1) to 500 μg L(-1). Phenotypic antibiotic resistance was quantified by culturing bacteria in the presence of different antibiotics, and genotypic resistance was determined using qPCR to quantify antibiotic resistance genes (ARGs). BAC at concentrations ranging from 0.1 μg L(-1) to 500 μg L(-1) was found to positively select for bacteria resistant to ciprofloxacin and sulfamethoxazole, and negatively select against bacteria with resistance to six other antibiotics. Exposure to BAC for 14 days increased the relative abundance of sul1 and bla(TEM). This study re-highlights the importance of employing both culture and non-culture-based techniques to identify selection for antibiotic resistance. The widespread use of BAC will likely impact antibiotic resistance profiles of bacteria in the environment, including in source waters used for drinking water, wastewater treatment plants, and natural waterways.202031767233
3720120.9995Urban wastewater as a conduit for pathogenic Gram-positive bacteria and genes encoding resistance to β-lactams and glycopeptides. The emergence and spread of clinical pathogens, antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs) in the environment pose a direct threat to human and animal health worldwide. In this study, we analyzed qualitatively and quantitatively urban sewage resistome for the occurrence of genes encoding resistance to β-lactams and glycopeptides in the genomes of culturable bacteria, as well as in the wastewater metagenome of the Central Wastewater Treatment Plant in Koziegłowy (Poland). Moreover, we estimated the presence of pathogenic Gram-positive bacteria in wastewater based on analysis of species-specific virulence genes in the wastewater metagenome. The results show that the final effluent contains alarm pathogens with particularly dangerous mechanisms of antibiotic resistance, including methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE). We also noticed that during the wastewater treatment, there is an increase in the frequency of MRSA and VRE. Furthermore, the results prove the effective removal of vanA, but at the same time show that wastewater treatment increases the relative abundance of mecA and virulence genes (groES and sec), indicating the presence of clinical pathogens E. faecalis and S. aureus in the effluent released to surface waters. We also observed an increase in the relative abundance of mecA and vanA genes already in the aeration tank, which suggests accumulation of contaminants affecting enhanced selection and HGT processes in the activated sludge. Moreover, we found a relation between the taxonomic composition and the copy number of ARGs as well as the presence of pathogens at various stages of wastewater treatment. The presence of clinically relevant pathogens, ARB, including multi-resistant bacteria, and ARGs in the effluent indicates that wastewater treatment plant play a key role in the existence of pathogens and antimicrobial resistance spreading pathway in the environment and human communities, which is a direct threat to public health and environmental protection.202133385807
3375130.9995Occurrence of antimicrobial agents, drug-resistant bacteria, and genes in the sewage-impacted Vistula River (Poland). Antimicrobial agents (antimicrobials) are a group of therapeutic and hygienic agents that either kill microorganisms or inhibit their growth. Their occurrence in surface water may reveal harmful effects on aquatic biota and challenge microbial populations. Recently, there is a growing concern over the contamination of surface water with both antimicrobial agents and multidrug-resistant bacteria. The aim of the study was the determination of the presence of selected antimicrobials at specific locations of the Vistula River (Poland), as well as in tap water samples originating from the Warsaw region. Analysis was performed using the liquid chromatography-electrospray ionization-tandem mass spectrometry method. In addition, the occurrence of drug-resistant bacteria and resistance genes was determined using standard procedures. This 2-year study is the first investigation of the simultaneous presence of antimicrobial agents, drug-resistant bacteria, and genes in Polish surface water. In Poland, relatively high concentrations of macrolides are observed in both surface and tap water. Simultaneous to the high macrolide levels in the environment, the presence of the erm B gene, coding the resistance to macrolides, lincosamides, and streptogramin, was detected in almost all sampling sites. Another ubiquitous gene was int1, an element of the 5'-conserved segment of class 1 integrons that encode site-specific integrase. Also, resistant isolates of Enterococcus faecium and Enterococcus faecalis and Gram-negative bacteria were recovered. Multidrug-resistant bacteria isolates of Gram-negative and Enterococcus were also detected. The results show that wastewater treatment plants (WWTP) are the main source of most antimicrobials, resistant bacteria, and genes in the aquatic environment, probably due to partial purification during wastewater treatment processes.201829235021
3433140.9995Effect of subinhibitory concentrations on the spreading of the ampicillin resistance gene bla(CMY-2) in an activated sludge microcosm. As the problem of multi-resistant bacteria grows a better understanding of the spread of antibiotic resistance genes is of utmost importance for society. Wastewater treatment plants contain subinhibitory concentrations of antibiotics and are thought to be hotspots for antibiotic resistance gene propagation. Here we evaluate the influence of sub-minimum inhibitory concentrations of antibiotics on the spread of resistance genes within the bacterial community in activated sludge laboratory-scale sequencing batch reactors. The mixed communities were fed two different ampicillin concentrations (500 and 5000 µg/L) and the reactors were run and monitored for 30 days. During the experiment the β-lactamase resistance gene bla(CMY-2) was monitored via qPCR and DNA samples were taken to monitor the effect of ampicillin on the microbial community. The relative copy number of bla(CMY-2) in the reactor fed with the sub-minimum inhibitory concentration of 500 µg/L ampicillin was spread out over a wider range of values than the control and 5000 µg/L ampicillin reactors indicating more variability of gene number in the 500 µg/L reactor. This result emphasises the problem of sub-minimum inhibitory concentrations of antibiotics in wastewater. High-throughput sequencing showed that continuous exposure to ampicillin caused a shift from a Bacteroidetes to Proteobacteria in the bacterial community. The combined use of qPCR and high-throughput sequencing showed that ampicillin stimulates the spread of resistance genes and leads to the propagation of microbial populations which are resistant to it.202539215485
3412150.9995Bacterial Resistance to β-Lactam Antibiotics in Municipal Wastewater: Insights from a Full-Scale Treatment Plant in Poland. This study investigated enzymatic and genetic determinants of bacterial resistance to β-lactam antibiotics in the biocenosis involved in the process of biological treatment of wastewater by activated sludge. The frequency of bacteria resistant to selected antibiotics and the activity of enzymes responsible for resistance to β-lactam antibiotics were estimated. The phenomenon of selection and spread of a number of genes determining antibiotic resistance was traced using PCR and gene sequencing. An increase in the percentage of bacteria showing resistance to β-lactam antibiotics in the microflora of wastewater during the treatment process was found. The highest number of resistant microorganisms, including multi-resistant strains, was recorded in the aeration chamber. Significant amounts of these bacteria were also present in treated wastewater, where the percentage of penicillin-resistant bacteria exceeded 50%, while those resistant to the new generation β-lactam antibiotics meropenem and imipenem were found at 8.8% and 6.4%, respectively. Antibiotic resistance was repeatedly accompanied by the activity of enzymes such as carbapenemases, metallo-β-lactamases, cephalosporinases and β-lactamases with an extended substrate spectrum. The activity of carbapenemases was shown in up to 97% of the multi-resistant bacteria. Studies using molecular biology techniques showed a high frequency of genes determining resistance to β-lactam antibiotics, especially the blaTEM1 gene. The analysis of the nucleotide sequences of blaTEM1 gene variants present in bacteria at different stages of wastewater treatment showed 50-100% mutual similarity of.202236557576
7304160.9995Nanofiltration 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
5353170.9995The influence of the autochthonous wastewater microbiota and gene host on the fate of invasive antibiotic resistance genes. The aim of this study was to assess the fate of invasive antibiotic resistance genes (ARG) discharged in wastewater. With this objective, antibiotic resistant bacteria (ARB) known to harbor specific ARG were inoculated in wastewater (hospital effluent, or municipal raw and treated wastewater) and in ultra-pure sterile water microcosms. Two sets of wastewater ARB isolates were used - set 1, Enterococcus faecalis, Acinetobacter johnsonii, Klebsiella pneumoniae and set 2, Enterococcus faecium, Acinetobacter johnsonii, Escherichia coli. Non-inoculated controls were run in parallel. Samples were collected at the beginning and at the end (15days) of the incubation period and the abundance of the genes 16S rRNA, intI1, bla(TEM) and vanA and the bacterial community composition were analyzed. In general, the genes bla(TEM) and vanA had lower persistence in wastewater and in ultra-pure water than the genes 16S rRNA or the class 1 integron integrase intI1. This effect was more pronounced in wastewater than in ultra-pure water, evidencing the importance of the autochthonous microbiota on the elimination of invasive ARG. Wastewater autochthonous bacterial groups most correlated with variations of the genes intI1, bla(TEM) and vanA were members of the classes Gammaproteobacteria, Bacilli or Bacteroidia. For bla(TEM), but not for vanA, the species of the ARB host was important to determine its fate. These are novel findings on the ecology of ARB in wastewater environments.201727697350
7799180.9995Combating Staphylococcus aureus and its methicillin resistance gene (mecA) with cold plasma. The increase in antibiotic resistance has become a global challenge to public health. In this study, an atmospheric cold plasma (ACP) system was applied for combating methicillin-resistant Staphylococcus aureus (MRSA) and its methicillin resistance gene (mecA) during food wastewater treatment. The plate count and flow cytometry methods were employed to estimate the damage in MRSA induced by plasma treatment. A quantitative real-time PCR (qPCR) method was used to assess the plasma-induced degradation of the mecA genes. The inactivation of MRSA and degradation of extracellular (e-) and intracellular (i-)mecA genes were investigated in phosphate buffered solution as a function of plasma exposure. A relatively low plasma influence of 0.12 kJ/cm(2) accounted for 5-log MRSA and 1.4-log e-mecA genes reduction, while only around 0.19-log degradation for i-mecA genes. As the plasma intensity was accumulated to 0.35 kJ/cm(2), the reduction of e- and i-mecA genes was increased to 2.6 and 0.8 logs, respectively. The degradation of i-mecA genes was much slower than that of e-mecA genes due to the protective effects of the outer envelopes or intracellular components against plasma. The matrix effect of wastewater effluents shielded both antibiotic resistance bacteria (ARB) and antibiotic resistance genes (ARGs) from plasma disinfection, which led to a lower degradation efficacy. Our results could support the development and optimization of plasma-based wastewater treatment.201830248853
7183190.9995Evaluation of five antibiotic resistance genes in wastewater treatment systems of swine farms by real-time PCR. Antibiotics are widely used in livestock for infection treatment and growth promotion. Wastes from animal husbandry are a potential environmental source of antibiotic-insensitive pathogens, and the removal efficiency of the resistance genotypes in current wastewater treatment plants (WWTPs) is unknown. In this study, quantitative PCR was used for evaluating antibiotic resistance genes in wastewater treatment processes. Six wastewater treatment plants in different swine farms were included in this study, and five antibiotic resistance genes (ARGs) were tested for each treatment procedure. All of the tested ARGs including tetA, tetW, sulI, sulII, and blaTEM genes were detected in six swine farms with considerable amounts. The results showed that antibiotic resistance is prevalent in livestock farming. The ARG levels were varied by wastewater treatment procedure, frequently with the highest level at anaerobic treatment tank and lowest in the activated sludge unit and the effluents. After normalizing the ARG levels to 16S rRNA gene copies, the results showed that ARGs in WWTP units fluctuated partly with the quantity of bacteria. Regardless of its importance in biodegradation, the anaerobic procedure may facilitate bacterial growth thus increasing the sustainability of the antibiotic resistance genotypes. After comparing the copy numbers in influx and efflux samples, the mean removal efficiency of ARGs ranged between 33.30 and 97.56%. The results suggested that treatments in the WWTP could partially reduce the spread of antibiotic-resistant bacteria, and additional procedures such as sedimentation may not critically affect the removal efficiency.201425064719