Antibiotic Resistance Genes Sul1 Research Articles

Native 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 NH4+-N and PO43--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 NH4+-N and PO43--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 blaTEM 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.

Nature-based solutions for antibiotics and antimicrobial resistance removal in tertiary wastewater treatment: Microbiological composition and risk assessment

This field-scale study evaluates the seasonal effectiveness of employing nature-based solutions (NBSs), particularly surface flow and horizontal subsurface flow constructed wetland configurations, as tertiary treatment technologies for the removal of antibiotics (ABs) and antibiotic resistance genes (ARGs) compared to a conventional treatment involving UV and chlorination. Out of the 21 monitored ABs, 13 were detected in the influent of three tertiary wastewater treatments, with concentrations ranging from 2 to 1218 ng·L-1. The ARGs sul1 and dfrA1 exhibited concentrations ranging from 1 × 105 to 9 × 106 copies/100 mL. NBSs were better at reducing ABs (average 69 to 88 %) and ARGs (2–3 log units) compared to the conventional tertiary system (average 36 to 39 % and no removal to 2 log units) in both seasons. Taxonomic compositions in influent water samples shifted from wastewater-impacted communities (Actinomycetota and Firmicutes) to a combination of plant rhizosphere-associated and river communities in NBS effluents (Alphaproteobacteria). In contrast, the conventional technology showed no substantial differences in community composition. Moreover, NBSs substantially reduced the ecotoxicological risk assessment (cumulative RQs). Furthermore, NBSs reduced the ecotoxicological risk (cumulative RQs) by an average of over 70 % across seasons, whereas the benchmark technology only achieved a 6 % reduction. In conclusion, NBSs present a robust alternative for minimizing the discharge of ABs and ARGs into surface water bodies.

Insights into the roles of biochar pores toward alleviating antibiotic resistance genes accumulation in biofiltration systems

Biofiltration systems would harbor and spread various antibiotic resistance genes (ARGs) when treating antibiotic micro-pollution, constituting a potential ecological risk. This study aimed to investigate the effects of biochar pores on ARG emergence and related microbial response mechanisms in bench-scale biofiltration systems. Results showed that biochar pores effectively reduced the absolute copies of the corresponding ARGs sul1 and sul2 by 54.1% by lowering the sorbed-SMX's bioavailability compared to non-porous anthracite. An investigation of antimicrobial resistomes revealed a considerable decrease in the abundance and diversity of ARGs and mobile gene elements. Metagenomic and metaproteomic analysis demonstrated that biochar pores induced the changeover of microbial defense strategy against SMX from blocking SMX uptake by EPS absorbing to SMX biotransformation. Microbial SOS response, antibiotic efflux pump, EPS secretion, and biofilm formation were decreased. Functions related to SMX biotransformation, such as sadABC-mediated transformation, xenobiotics degradation, and metabolism, were significantly promoted.

Microalgal Systems, a Green Solution for Wastewater Conventional Pollutants Removal, Disinfection, and Reduction of Antibiotic Resistance Genes Prevalence?

The low-efficiency rate of urban wastewater (UWW) treatment generates tons of discharged water with a high concentration of pollutants, pathogens and antibiotic-resistance genes (ARGs). Microalgal systems may be a green alternative to be implemented as a UWW polishing treatment. This study assessed the ability of Chlorella vulgaris and UWW autochthonous microalgal species (AMS) to simultaneously remove PO4–P, and reduce the proliferation of coliforms and ARGs. AMS seems to be more promising due to: (i) the higher specific growth rate, μmax (0.687 ± 0.065 d−1); (ii) efficient PO4–P removal (92.62 ± 0.10%); (iii) faster reduction of coliforms proliferation achieving concentrations below the limits of quantification (6 d); (iv) the reduction of intl1 and the ARGs sul1 and blaTEM abundance in ca. of 70.4%, 69.2%, and 75.7%, respectively (9 d); and (v) the additional reduction of these genes in ca. of 97.1%, 94.2%, and 99.9%, respectively, after 5 d storage in the dark and at room temperature. Results also revealed that the high pH values in both microalgal systems (due to microalgal growth) were highly correlated with a reduction in the proliferation of coliforms, including Escherichia coli. In conclusion, using AMS as a final polishing treatment of UWW seems to be very promising.

Global dispersal and potential sources of antibiotic resistance genes in atmospheric remote depositions.

Antibiotic resistance has become a major Global Health concern and a better understanding on the global spread mechanisms of antibiotic resistant bacteria (ARB) and intercontinental ARB exchange is needed. We measured atmospheric depositions of antibiotic resistance genes (ARGs) by quantitative (q)PCR in rain/snow collected fortnightly along 4 y. at a remote high mountain LTER (Long-Term Ecological Research) site located above the atmospheric boundary layer (free troposphere). Bacterial composition was characterized by 16S rRNA gene sequencing, and air mass provenances were determined by modelled back trajectories and rain/snow chemical composition. We hypothesize that the free troposphere may act as permanent reservoir and vector for ARB and ARGs global dispersal. We aimed to i) determine whether ARGs are long-range intercontinental and persistently dispersed through aerosols, ii) assess ARGs long-term atmospheric deposition dynamics in a remote high mountain area, and iii) unveil potential diffuse ARGs pollution sources. We showed that the ARGs sul1 (resistance to sulfonamides), tetO (resistance to tetracyclines), and intI1 (a proxy for horizontal gene transfer and anthropogenic pollution) were long-range and persistently dispersed in free troposphere aerosols. Major depositions of tetracyclines resistance matched with intensification of African dust outbreaks. Potential ARB mostly traced their origin back into agricultural soils. Our study unveils that air masses pathways are shaping ARGs intercontinental dispersal and global spread of antibiotic resistances, with potential predictability for interannual variability and remote deposition rates. Because climate regulates aerosolization and long-range air masses movement patterns, we call for a more careful evaluation of the connections between land use, climate change and ARB long-range intercontinental dispersal.

Anaerobic Digestion and Removal of Sulfamethoxazole, Enrofloxacin, Ciprofloxacin and Their Antibiotic Resistance Genes in a Full-Scale Biogas Plant.

Anaerobic digestion is one of the best ways to re-use animal manure and agricultural residues, through the production of combustible biogas and digestate. However, the use of antibiotics for preventing and treating animal diseases and, consequently, their residual concentrations in manure, could introduce them into anaerobic digesters. If the digestate is applied as a soil fertilizer, antibiotic residues and/or their corresponding antibiotic resistance genes (ARGs) could reach soil ecosystems. This work investigated three common soil emerging contaminants, i.e., sulfamethoxazole (SMX), ciprofloxacin (CIP), enrofloxacin (ENR), their ARGs sul1, sul2, qnrS, qepA, aac-(6′)-Ib-cr and the mobile genetic element intI1, for one year in a full scale anaerobic plant. Six samplings were performed in line with the 45-day hydraulic retention time (HRT) of the anaerobic plant, by collecting input and output samples. The overall results show both antibiotics and ARGs decreased during the anaerobic digestion process. In particular, SMX was degraded by up to 100%, ENR up to 84% and CIP up to 92%, depending on the sampling time. In a similar way, all ARGs declined significantly (up to 80%) in the digestate samples. This work shows how anaerobic digestion can be a promising practice for lowering antibiotic residues and ARGs in soil.

Antibiotic resistance gene distribution in agricultural fields and crops. A soil-to-food analysis

Despite the social concern about the generalization of antibiotic resistance hotspots worldwide, very little is known about the contribution of different potential sources to the global risk. Here we present a quantitative analysis of the distribution of Antibiotic Resistance Genes (ARGs) in soil, rhizospheric soil, roots, leaves and beans in tomato, lettuce and broad beans crops (165 samples in total), grown in nine commercial plots distributed in four geographical zones in the vicinity of Barcelona (North East Spain). We also analyzed five soil samples from a nearby forest, with no record of agricultural activities. DNA samples were analyzed for their content in the ARGs sul1, tetM, qnrS1, blaCTX-M-32, blaOXA-58, mecA, and blaTEM, plus the integron intI1, using qPCR methods. In addition, soil microbiomes from the different plots were analyzed by amplicon-targeted 16S rRNA gene sequencing. Our data show a decreasing gradient of ARG loads from soil to fruits and beans, the latter showing only from 0.1 to 0.01% of the abundance values in soil. The type of crop was the main determinant for both ARG distribution and microbiome composition among the different plots, with minor contributions of geographic location and irrigation water source. We propose that soil amendment and/or fertilization, more than irrigation water, are the main drivers of ARG loads on the edible parts of the crop, and that they should therefore be specifically controlled.

Spatially resolved abundances of antibiotic resistance genes and intI1 in wastewater treatment biofilms.

Attached growth bioprocesses that use biofilms to remove organic matter or nutrients from wastewater are known to harbor antibiotic resistance genes (ARGs). Biofilms in these processes are spatially heterogeneous, but little is known about depth stratification of ARGs in complex, mixed culture biofilms. To address this knowledge gap, we used an experimental approach combining cryosectioning and quantitative polymerase chain reaction to quantify the spatial distribution of three ARGs (sul1, ermB, and qnrS) and the class 1 integron-integrase gene intI1 in biofilms from a lab-scale rotating annular reactor fed with synthetic wastewater. We also used high throughput 16S ribosomal RNA (rRNA) gene sequencing to characterize community structure with depth in biofilms. The ARG sul1 and the integron-integrase gene intI1 were found in higher abundances in upper layers of biofilm near the fluid-biofilm interface than in lower layers and exhibited significant correlations between the distance from substratum and gene abundances. The genes ermB and qnrS were present in comparatively low relative abundances. Microbial community structure varied significantly by date of sampling and distance from the substratum. These findings highlight the genetic and taxonomic heterogeneity with distance from substratum in wastewater treatment biofilms and show that sul1 and intI1 are particularly abundant near fluid-biofilm interfaces where cells are most likely to detach and flow into downstream portions of treatment systems and can ultimately be released into the environment through effluent.

Viability-based quantification of antibiotic resistance genes and human fecal markers in wastewater effluent and receiving waters

Antibiotic resistance is a public health issue with links to environmental sources of antibiotic resistance genes (ARGs). ARGs from nonviable sources may pose a hazard given the potential for transformation whereas ARGs in viable sources may proliferate during host growth or conjugation. In this study, ARGs in the effluent from three municipal wastewater treatment plants (WWTPs) and the receiving surface waters were investigated using a viability-based qPCR technique (vPCR) with propidium monoazide (PMA). ARGs sul1, tet(G), and blaTEM, fecal indicator marker BacHum, and 16S rRNA gene copies/mL were found to be significantly lower in viable-cells than in total concentrations for WWTP effluent. Viable-cell and total gene copy concentrations were similar in downstream samples except for tet(G). Differences with respect to season in the prevalence of nonviable ARGs in surface water or WWTP effluent were not observed. The results of this study indicate that qPCR may overestimate viable-cell ARGs and fecal indicator genes in WWTP effluent but not necessarily in the surface water >1.8 km downstream.

Antibiotic Resistance in Airborne Bacteria Near Conventional and Organic Beef Cattle Farms in California, USA

Levels of antibiotic resistance genes (ARGs) and the fractions of antibiotic resistant bacteria (ARB) among culturable heterotrophic bacteria were compared in outdoor air near conventional (n = 3) and organic (n = 3) cattle rearing facilities. DNA extracts from filters taken from 18 locations were analyzed by quantitative polymerase chain reaction (qPCR) for five ARGs. At the reference (non-agricultural) site, all genes were below detection. ARGs sul1, bla SHV, erm(B), and bla TEM were more frequently detected and at higher levels (up to 870 copies m−3 for bla SHV and 750 copies m−3 for sul1) near conventional farms compared to organic locations while the opposite was observed for erm(F) (up to 210 copies m−3). Isolates of airborne heterotrophic bacteria (n = 1295) collected from the sites were tested for growth in the presence of six antibiotics. By disk diffusion on a subset of isolates, the fractions of ARB were higher for conventional sites compared to organic farms for penicillin (0.9 versus 0.63), cloxacillin (0.74 versus 0.23), cefoperazone (0.58 versus 0.14), and sulfamethazine (0.50 versus 0.33) for isolates on nutrient agar. All isolates’ ΔA600pres/ΔA600abs were measured for each of the six tested antibiotics; isolates from farms downwind of organic sites had a lower average ΔA600pres/ΔA600abs for most antibiotics. In general, all three analyses used to indicate microbial resistance to antibiotics showed increases in air samples nearby conventional versus organic cattle rearing facilities. Regular surveillance of airborne ARB and ARGs near conventional and organic beef cattle farms is suggested.

Investigation of the potential of a Membrane BioReactor followed by solar Fenton oxidation to remove antibiotic-related microcontaminants

There is limited research regarding alternative technologies of controlling the presence of antibiotic residues and antibiotic tolerance/resistance in urban wastewater treatment plants (UWTPs). This study deals with the efficiency of a Membrane BioReactor (MBR) integrated with solar Fenton oxidation for the removal of selected antibiotic-related microcontaminants, at a pilot scale. More specifically, the aspects examined in this study included: (i) the removal of three antibiotics, namely sulfamethoxazole (SMX), erythromycin (ERY) and clarithromycin (CLA), (ii) the prevalence of total and antibiotic-tolerant bacteria, (iii) the total DNA and antibiotic resistance genes (ARG) removal efficiency of the integrated process, as well as the abundance of taxon-specific markers. The quantitative examination of the presence of antibiotic residues in the MBR-treated effluent revealed a concentration of SMX of 5.5ngL−1, of CLA of 7.2ngL−1, while ERY concentration was below the limit of detection (LOD). Due to the low antibiotic concentrations in the MBR effluent, spiking of the examined antibiotics (100μgL−1) was done to examine their photo-persistence after solar Fenton oxidation. SMX and ERY concentrations were below the LOD after t30W,n=119.2min, while CLA was reduced by 84%. Total and antibiotic-tolerant cultivable bacteria Escherichia coli and Klebsiella spp. were completely inactivated. On the other hand, there was repair of Pseudomonas aeruginosa observed, with 2CFU 100mL−1 growing on the selective media 24h after solar Fenton oxidation. The total DNA concentration was reduced by 97%, while in the remaining total DNA determined after treatment, the Enterococcus spp. specific gene marker (3.9log10 CE 100ng−1 DNA), and the ARG sul1 and ermB (1.56 and 1.53log10 CE 100ng−1 DNA, respectively) were still present, indicating the further challenge of their removal.

Removal of antibiotics and antibiotic resistance genes in rural wastewater by an integrated constructed wetland

Integrated constructed wetlands (ICWs) are regarded as one of the most important removal technology for pollutants in rural domestic wastewaters. This study investigated the efficiency of an ICW consisting of a regulating pool, four surface and subsurface flow-constructed wetlands, and a stabilization unit for removing antibiotics and antibiotic resistance genes (ARGs) from rural domestic wastewaters. The results showed that antibiotics leucomycin, ofloxacin, lincomycin, and sulfamethazine, and ARGs sul1, sul2, tetM, and tetO were the predominant antibiotics and ARGs in the influent, respectively. The ICW system could significantly reduce most of the detected antibiotics and ARGs with their aqueous removal rates of 78 to 100 % and >99 %, respectively. Based on the measured concentrations, the total pollution loadings of antibiotics were 3,479 μg/day in the influent and 199 μg/day in the final effluent. Therefore, constructed wetlands could be a promising technology for rural wastewater in removing contaminants such as antibiotics and ARGs.