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713300.9967Prevalence of antibiotic resistance genes in bacterial communities associated with Cladophora glomerata mats along the nearshore of Lake Ontario. The alga Cladophora glomerata can erupt in nuisance blooms throughout the lower Great Lakes. Since bacterial abundance increases with the emergence and decay of Cladophora, we investigated the prevalence of antibiotic resistance (ABR) in Cladophora-associated bacterial communities up-gradient and down-gradient from a large sewage treatment plant (STP) on Lake Ontario. Although STPs are well-known sources of ABR, we also expected detectable ABR from up-gradient wetland communities, since they receive surface run-off from urban and agricultural sources. Statistically significant differences in aquatic bacterial abundance and ABR were found between down-gradient beach samples and up-gradient coastal wetland samples (ANOVA, Holm-Sidak test, p < 0.05). Decaying and free-floating Cladophora sampled near the STP had the highest bacterial densities overall, including on ampicillin- and vancomycin-treated plates. However, quantitative polymerase chain reaction analysis of the ABR genes ampC, tetA, tetB, and vanA from environmental communities showed a different pattern. Some of the highest ABR gene levels occurred at the 2 coastal wetland sites (vanA). Overall, bacterial ABR profiles from environmental samples were distinguishable between living and decaying Cladophora, inferring that Cladophora may control bacterial ABR depending on its life-cycle stage. Our results also show how spatially and temporally dynamic ABR is in nearshore aquatic bacteria, which warrants further research.201728192677
727610.9963Antibiotic resistance in urban and hospital wastewaters and their impact on a receiving freshwater ecosystem. The main objective of this study was to investigate the antibiotic resistance (AR) levels in wastewater (WW) and the impact on the receiving river. Samples were collected once per season over one year in the WW of a hospital, in the raw and treated WW of two wastewater treatment plants (WWTPs), as well as upstream and downstream from the release of WWTPs effluents into the Zenne River (Belgium). Culture-dependent methods were used to quantify Escherichia coli and heterotrophic bacteria resistant to amoxicillin, sulfamethoxazole, nalidixic acid and tetracycline. Six antibiotic resistance genes (ARGs) were quantified in both particle-attached (PAB) and free-living (FLB) bacteria. Our results showed that WWTPs efficiently removed antibiotic resistant bacteria (ARB) regardless of its AR profile. The ARGs levels were the highest in the hospital WW and were significantly reduced in both WWTPs. However, ARB and ARGs abundances significantly increased into the Zenne River downstream from the WWTPs outfalls. The variation in the relative abundance of ARGs through WW treatment differed depending on the WWTP, fraction, and gene considered. The sul1 and sul2 genes in PAB fraction showed significantly higher relative abundances in the effluent compared to the influent of both WWTPs. This study demonstrated that WWTPs could be hotspots for AR spread with significant impacts on receiving freshwater ecosystems. This was the first comprehensive study investigating at the same time antibiotics occurrence, fecal bacteria indicators, heterotrophic bacterial communities, and ARGs (distinguishing PAB and FLB) to assess AR levels in WW and impacts on the receiving river.201829730567
708720.9961Antibiotic resistance in shellfish and major inland pollution sources in the drainage basin of Kamak Bay, Republic of Korea. Shellfish-growing areas in marine environments are affected by pollutants that mainly originate from land, including streams, domestic wastewater, and the effluents of wastewater treatment plants (WWTPs), which may function as reservoirs of antibiotic-resistant bacteria (ARB) and antibiotic-resistance genes (ARGs). The objective of this study was to identify the occurrence and distribution of antibiotic resistance at five oyster sampling sites and 11 major inland pollution sources in the drainage basin of Kamak Bay, Republic of Korea. Culture-based methods were used to estimate the diversity and abundance of antibiotic-resistant Escherichia coli strains isolated from oysters and major inland pollution sources. The percentages of ARB and multiple antibiotic resistance index values were significantly high in discharge water from small fishing villages without WWTPs. However, the percentages of antibiotic-resistant E. coli isolates from oysters were low, as there was no impact from major inland pollutants. Fourteen ARGs were also quantified from oysters and major inland pollution sources. Although most ARGs except for quinolones were widely distributed in domestic wastewater discharge and effluent from WWTPs, macrolide resistance genes (ermB and msrA) were detected mainly from oysters in Kamak Bay. This study will aid in tracking the sources of antibiotic contamination in shellfish to determine the correlation between shellfish and inland pollution sources.202134226964
349830.9960Comparative study on the bacterial diversity and antibiotic resistance genes of urban landscape waters replenished by reclaimed water and surface water in Xi'an, China. Pathogenic bacteria and antibiotic resistance genes (ARGs) in urban landscape waters may pose a potential threat to human health. However, the investigation of their occurrence in the urban landscape waters replenished by reclaimed water (RW) and surface water (SW) is still insufficient. The water samples collected from six urban landscape waters replenished by RW or SW were used to analyze bacterial diversity using high-throughput sequencing of 16S rRNA gene and to detect 18 ARGs and 2 integron-integrase genes by means of quantitative PCR array. Results indicated that Proteobacteria was the dominant phylum in all six urban landscape waters. The bacterial species richness was lower in urban landscape waters replenished by RW than that by SW. Sulfonamide resistance genes (sulI and sulIII) were the major ARGs in these urban landscape waters. No significant difference in the relative abundance of sulfonamide resistance genes, tetracycline resistance genes, and most of beta-lactam resistance genes was observed between RW-replenished and SW-replenished urban landscape waters. By contrast, the relative abundance of bla(ampC) gene and qnrA gene in RW-replenished urban landscape waters was significantly higher than that in SW-replenished urban landscape waters (p < 0.05), which suggested that use of RW may increase the amount of specific ARGs to urban landscape waters. Interestingly, among six urban landscape waters, RW-replenished urban landscape waters had a relatively rich variety of ARGs (12-15 of 18 ARGs) but a low relative abundance of ARGs (458.90-1944.67 copies/16S × 10(6)). The RW replenishment was found to have a certain impact on the bacterial diversity and prevalence of ARGs in urban landscape waters, which provide new insight into the effect of RW replenishment on urban landscape waters.202133786766
709140.9960Abundance of Class 1 Integron-Integrase and Sulfonamide Resistance Genes in River Water and Sediment Is Affected by Anthropogenic Pressure and Environmental Factors. In this study, we determined the presence of class 1 integron-integrase gene in culturable heterotrophic bacteria isolated from river water and sediment sampled upstream and downstream of a wastewater treatment plant effluent discharge. Moreover, we quantified intI1 and sulfonamide resistance genes (sul1 and sul2) in the water and sediment using qPCR. There was no correlation between the results from water and sediment samples, which suggests integron-containing bacteria are differentially retained in these two environmental compartments. The discharge of treated wastewater significantly increased the frequency of intI1 among culturable bacteria and the gene copy number in river water, and increased the number of sul1 genes in the sediment. We also observed seasonal differences in the frequency of the class 1 integron-integrase gene among culturable heterotrophs as well as intI1 copy number in water, but not in sediment. The results suggest that the abundance of class 1 integrons in aquatic habitat depends on anthropogenic pressure and environmental factors.201627599709
709850.9960Antibiotic resistance along an urban river impacted by treated wastewaters. Urban rivers are impacted ecosystems which may play an important role as reservoirs for antibiotic-resistant (AR) bacteria. The main objective of this study was to describe the prevalence of antibiotic resistance along a sewage-polluted urban river. Seven sites along the Zenne River (Belgium) were selected to study the prevalence of AR Escherichia coli and freshwater bacteria over a 1-year period. Culture-dependent methods were used to estimate E. coli and heterotrophic bacteria resistant to amoxicillin, sulfamethoxazole, nalidixic acid and tetracycline. The concentrations of these four antibiotics have been quantified in the studied river. The antibiotic resistance genes (ARGs), sul1, sul2, tetW, tetO, blaTEM and qnrS were also quantified in both particle-attached (PAB) and free-living (FLB) bacteria. Our results showed an effect of treated wastewaters release on the spread of antibiotic resistance along the river. Although an increase in the abundance of both AR E. coli and resistant heterotrophic bacteria was observed from upstream to downstream sites, the differences were only significant for AR E. coli. A significant positive regression was also found between AR E. coli and resistant heterotrophic bacteria. The concentration of ARGs increased from upstream to downstream sites for both particle-attached (PAB) and free-living bacteria (FLB). Particularly, a significant increase in the abundance of four among six ARGs analyzed was observed after crossing urban area. Although concentrations of tetracycline significantly correlated with tetracycline resistance genes, the antibiotic levels were likely too low to explain this correlation. The analysis of ARGs in different fractions revealed a significantly higher abundance in PAB compared to FLB for tetO and sul2 genes. This study demonstrated that urban activities may increase the spread of antibiotic resistance even in an already impacted river.201829453174
709660.9960Sulfonamide and tetracycline resistance genes in total- and culturable-bacterial assemblages in South African aquatic environments. Antibiotic resistant bacteria are ubiquitous in the natural environment. The introduction of effluent derived antibiotic resistance genes (ARGs) into aquatic environments is of concern in the spreading of genetic risk. This study showed the prevalence of sulfonamide and tetracycline resistance genes, sul1, sul2, sul3, and tet(M), in the total bacterial assemblage and colony forming bacterial assemblage in river and estuarine water and sewage treatment plants (STP) in South Africa. There was no correlation between antibiotic concentrations and ARGs, suggesting the targeted ARGs are spread in a wide area without connection to selection pressure. Among sul genes, sul1 and sul2 were major genes in the total (over 10(-2) copies/16S) and colony forming bacteria assemblages (∼10(-1) copies/16S). In urban waters, the sul3 gene was mostly not detectable in total and culturable assemblages, suggesting sul3 is not abundant. tet(M) was found in natural assemblages with 10(-3) copies/16S level in STP, but was not detected in colony forming bacteria, suggesting the non-culturable (yet-to-be cultured) bacterial community in urban surface waters and STP effluent possess the tet(M) gene. Sulfamethoxazole (SMX) resistant (SMX(r)) and oxytetracycline (OTC) resistant (OTC(r)) bacterial communities in urban waters possessed not only sul1 and sul2 but also sul3 and tet(M) genes. These genes are widely distributed in SMX(r) and OTC(r) bacteria. In conclusion, urban river and estuarine water and STP effluent in the Durban area were highly contaminated with ARGs, and the yet-to-be cultured bacterial community may act as a non-visible ARG reservoir in certain situations.201526300864
708870.9959Small-scale wastewater treatment plants as a source of the dissemination of antibiotic resistance genes in the aquatic environment. Wastewater treatment plants (WWTPs) are significant source of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs), which can spread further in the environment by reaching rivers together with effluents discharged from WWTPs. In this study untreated and treated wastewater (UWW, TWW), upstream and downstream river water (URW, DRW) were collected from 4 WWTPs, in the winter and autumn seasons. The occurrence of ARB resistant to beta-lactams and tetracyclines as well as the presence of antibiotics from these classes were analysed in water and wastewater samples. Additionally, the amounts of 12 ARGs, 2 genes of mobile genetic elements (MGEs), gene uidA identifying E. coli and 16S rRNA were also determined. Resistance to beta-lactams prevailed among ARB in water and wastewater samples (constituting 82-88% of total counts of bacteria). The dominant genes in water and wastewater samples were bla(TEM), tetA, sul1. The gene bla(OXA) demonstrated high variability of its concentration in samples collected in both seasons. Despite the high per cent reduction of ARB and ARGs concentration observed during the wastewater treatment processes, their large quantities are still transmitted into the environment. The research focuses on WWTPs' role in the dissemination of ARGs and MGEs in the aquatic environment.202031561123
728080.9959Microbial communities of biofilms developed in a chlorinated drinking water distribution system: A field study of antibiotic resistance and biodiversity. Antibiotic resistance and biodiversity were investigated in microbial communities attached to inner surfaces of water supply fittings in a chlorinated drinking water distribution system (DWDS) supplied by two independent water treatment plants (WTPs) drawing the same source water. The investigation of the effect of the season, the applied water treatment technology, and type, material, and age of water supply fittings on both antibiotic resistance and biodiversity in biofilms involved collection of tubercles during summer and winter seasons throughout the DWDS. A total of 16 samples were collected (8 per season) from areas supplied by two independent WTPs. Culturable aerobic antibiotic resistant bacteria (ARB) proved more prevalent in summer. Various antibiotic resistance genes (ARGs) were detected, confirming the role of biofilms as ARGs reservoirs, but the abundances of quantified genes (sulI, ermB, qacEΔ1, intI1) were low (a range of 202133610999
727790.9959Occurrence and persistence of carbapenemases genes in hospital and wastewater treatment plants and propagation in the receiving river. This study aims to investigate the prevalence of clinically relevant carbapenemases genes (bla(KPC), bla(NDM) and bla(OXA-48)) in water samples collected over one-year period from hospital (H), raw and treated wastewater of two wastewater treatment plants (WWTPs) as well as along the Zenne River (Belgium). The genes were quantified in both particle-attached (PAB) and free-living (FLB) bacteria. Our results showed that absolute abundances were the highest in H waters. Although absolute abundances were significantly reduced in WWTP effluents, the relative abundance (normalized per 16S rRNA) was never lowered through wastewater treatment. Particularly, for the PAB the relative abundances were significantly higher in the effluents respect to the influents of both WWTPs for all the genes. The absolute abundances along the Zenne River increased from upstream to downstream, peaking after the release of WWTPs effluents, in both fractions. Our results demonstrated that bla(KPC), bla(NDM) and bla(OXA-48) are widely distributed in the Zenne as a consequence of chronic discharge from WWTPs. To conclude, the levels of carbapenemases genes are significantly lower than other genes conferring resistance to more widely used antibiotics (analyzed in previous studies carried out at the same sites), but could raise up to the levels of high prevalent resistance genes.201829960932
3542100.9958Fecal indicators, pathogens, antibiotic resistance genes, and ecotoxicity in Galveston Bay after Hurricane Harvey. Unprecedented rainfall after Hurricane Harvey caused a catastrophic flood in the southern coast of Texas, and flushed significant floodwater and sediments into Galveston Bay, the largest estuary along the Texas Gulf Coast. This study investigated the immediate and long-term (6 months post-Harvey) fecal indicators, pathogenic bacteria, antibiotic resistance genes (ARGs), and ecotoxicity in the Galveston Bay. Dramatic decrease of salinity profile to zero, increased levels of fecal indicator bacteria and pathogenic bacteria, and detection of various ARGs were observed in the water and sediment samples collected 2 weeks post-Harvey. High levels of Bla(TEM) and cytotoxicity measured by yeast bioluminescent assay (BLYR) were also observed especially near the river mouths. While Vibrio spp. was dominant in water, much higher abundance of fecal indicator bacteria and pathogen were detected in the sediments. A decreasing trend of Bla(TEM) and cytotoxicity was observed in March 2018 samples, suggesting the Bay has returned to its pre-hurricane conditions 6 months post-Harvey. Interestingly, the abundance of fecal indicator bacteria and pathogens were shifted dramatically according to high-streamflow and low-streamflow seasons in the Bay. The data are useful to construct the model of risk assessment in coastal estuaries system and predict the effects of extreme flooding events in the future.202133445049
7291110.9958Accumulation of clinically relevant antibiotic-resistance genes, bacterial load, and metals in freshwater lake sediments in Central Europe. Wastewater treatment plants (WWTP) receive the effluents from various sources (communities, industrial, and hospital effluents) and are recognized as reservoir for antibiotic-resistance genes (ARGs) that are associated with clinical pathogens. The aquatic environment is considered a hot-spot for horizontal gene transfer, and lake sediments offer the opportunity for reconstructing the pollution history and evaluating the impacts. In this context, variation with depth and time of the total bacterial load, the abundance of faecal indicator bacteria (FIB; E. coli and Enterococcus spp. (ENT)), Pseudomonas spp., and ARGs (blaTEM, blaSHV, blaCTX-M, blaNDM, and aadA) were quantified in sediment profiles of different parts of Lake Geneva using quantitative PCR. The abundance of bacterial marker genes was identified in sediments contaminated by WWTP following eutrophication of the lake. Additionally, ARGs, including the extended-spectrum ß-lactam- and aminoglycoside-resistance genes, were identified in the surface sediments. The ARG and FIB abundance strongly correlated (r ≥ 0.403, p < 0.05, n = 34) with organic matter and metal concentrations in the sediments, indicating a common and contemporary source of contamination. The contamination of sediments by untreated or partially treated effluent water can affect the quality of ecosystem. Therefore, the reduction of contaminants from the source is recommended for further improvement of water quality.201525933054
7097120.9958Markers Specific to Bacteroides fragilis Group Bacteria as Indicators of Anthropogenic Pollution of Surface Waters. The aim of this study was to evaluate the applicability of markers specific to Bacteroides fragilis group (BFG) bacteria as indicators of anthropogenic pollution of surface waters. In addition, the impact of wastewater treatment plants (WWTPs) on the spread of genes specific to fecal indicator bacteria and genes encoding antimicrobial resistance in water bodies was also determined. Samples of hospital wastewater (HWW), untreated wastewater (UWW), and treated wastewater (TWW) evacuated from a WWTP were collected, and samples of river water were taken upstream (URW) and downstream (DRW) from the wastewater discharge point to determine, by qPCR, the presence of genes specific to BFG, Escherichia coli and Enterococcus faecalis, and the abundance of 11 antibiotic resistance genes (ARGs) and two integrase genes. The total number of bacterial cells (TCN) in the examined samples was determined by fluorescence in situ hybridization (FISH). Genes specific to BFG predominated among the analyzed indicator microorganisms in HWW, and their copy numbers were similar to those of genes specific to E. coli and E. faecalis in the remaining samples. The abundance of genes specific to BFG was highly correlated with the abundance of genes characteristic of E. coli and E. faecalis, all analyzed ARGs and intI genes. The results of this study indicate that genes specific to BFG can be used in analyses of human fecal pollution, and as indicators of environmental contamination with ARGs. A significant increase in the copy numbers of genes specific to BFG, E. coli, and seven out of the 11 analyzed ARGs was noted in samples of river water collected downstream from the wastewater discharge point, which suggests that WWTPs are an important source of these genes in riparian environments.202033003501
7295130.9958Dissemination of antibiotic resistance in receiving environments under a changing climate: A modeling exercise. Antibiotic resistance in rivers has become a global problem, particularly due to the discharge of wastewater treatment plant (WWTP) effluents into these systems. These effluents contain residual antibiotics, antibiotic-resistance genes (ARGs), and antibiotic-resistant bacteria (ARB). While watershed-scale models are commonly used to address other water quality issues, they have not typically been used to address antibiotic resistance. In this study, we present a new model called SWAT-ARB (SWAT- Antibiotic-Resistant Bacteria) that can simulate antibiotic resistance in E. coli at the watershed scale. SWAT-ARB is an adaptation of the widely-used SWAT (Soil and Water Assessment Tool) model, which is a physically-based, watershed-scale hydrological model. We used SWAT-ARB to study the receiving environments of WWTPs in the Adyar River basin in India, Crab Creek in the United States, and the Upper Viskan basin in Sweden. We analyzed the simulations of resistant fractions (the ratio of resistant E. coli concentration to total E. coli concentration) in the streamflow at different flow levels. We also examined the long-term trends of resistant fractions to understand how rising temperatures may impact resistance. We found that in the Adyar and Crab Creek basins, the resistant fractions were largely influenced by temperature rather than flow and wash-off processes, while in the Upper Viskan basin, the resistant fractions were affected by both temperature and flow conditions. In a simulation where we only increased temperatures by 2 °C in the bacteria sub-routine, we found that the Adyar basin showed a decrease in resistant fractions of up to 17 % in dry conditions, while Crab Creek showed increases of 17.5-24.1 % and Upper Viskan showed increases of 4.6-33.5 % across flow classes. Under future climate scenarios (SSP 2-4.5 and SSP 5-8.5), Adyar's resistant fractions decreased by up to 55.5 % as temperatures approached the bacterial growth inhibition threshold, while Crab Creek's resistant fractions increased by up to 175 % as temperatures remained within the optimal 10-20 °C growth range. Our results suggest that the SWAT-ARB model could be further improved by incorporating temperature-dependent parameters into the resistance simulation component.202540743959
7223140.9957Characterization of metal resistance genes carried by waterborne free-living and particle-attached bacteria in the Pearl River Estuary. Toxic metals can substantially change the bacterial community and functions thereof in aquatic environments. Herein, metal resistance genes (MRGs) are the core genetic foundation for microbial responses to the threats of toxic metals. In this study, waterborne bacteria collected from the Pearl River Estuary (PRE) were separated into the free-living bacteria (FLB) and particle-attached bacteria (PAB), and analyzed using metagenomic approaches. MRGs were ubiquitous in the PRE water and mainly related to Cu, Cr, Zn, Cd and Hg. The levels of PAB MRGs in the PRE water ranged from 8.11 × 10(9) to 9.93 × 10(12) copies/kg, which were significantly higher than those of the FLB (p < 0.01). It could be attributed to a large bacterial population attached on the suspended particulate matters (SPMs), which was evidenced by a significant correlation between the PAB MRGs and 16S rRNA gene levels in the PRE water (p < 0.05). Moreover, the total levels of PAB MRGs were also significantly correlated with those of FLB MRGs in the PRE water. The spatial pattern of MRGs of both FLB and PAB exhibited a declining trend from the low reaches of the PR to the PRE and on to the coastal areas, which was closely related to metal pollution degree. MRGs likely carried by plasmids were also enriched on the SPMs with a range from to 3.85 × 10(8) to 3.08 × 10(12) copies/kg. MRG profiles and taxonomic composition of the predicted MRG hosts were significantly different between the FLB and PAB in the PRE water. Our results suggested that FLB and PAB could behave differential response to heavy metals in the aquatic environments from the perspective of MRGs.202337028791
7781150.9957Untreated HWWs Emerged as Hotpots for ARGs. Hospital wastewaters (HWWs) are reported to be hotspots for antibiotics and antibiotic-resistant bacteria. However, limited information involves the impact of these effluents on dissemination of antibiotic-resistance genes (ARGs). In this study, therefore, seasonally collected HWWs were monitored for overall bacterial load and seven ARGs aadA, tetA, cmlA, sul1, qnrS, ermB and bla (CTX-M )by using quantitative polymerase chain reaction method. Overall bacterial 16S rRNA copy number was found to be the lowest in winter with 10(3 )copy number/mL, while the highest copy number, with 10(5 )copy number/mL, was observed in both summer and spring. All hospitals tested displayed similar seasonal ARG copy number profile of aadA > tetA > cmlA ≈ sul1 > ermB ≈ qnrS > bla (CTX-M). The results indicated that untreated HWWs were hotspots for ARGs and required attention before discharging into public sewer.202031965225
7349160.9957Bacterial and DNA contamination of a small freshwater waterway used for drinking water after a large precipitation event. Sewage contamination of freshwater occurs in the form of raw waste or as effluent from wastewater treatment plants (WWTP's). While raw waste (animal and human) and under-functioning WWTP's can introduce live enteric bacteria to freshwater systems, most WWTP's, even when operating correctly, do not remove bacterial genetic material from treated waste, resulting in the addition of bacterial DNA, including antibiotic resistance genes, into water columns and sediment of freshwater systems. In freshwater systems with both raw and treated waste inputs, then, there will be increased interaction between live sewage-associated bacteria (untreated sewage) and DNA contamination (from both untreated and treated wastewater effluent). To evaluate this understudied interaction between DNA and bacterial contamination in the freshwater environment, we conducted a three-month field-based study of sewage-associated bacteria and genetic material in water and sediment in a freshwater tributary of the Hudson River (NY, USA) that supplies drinking water and receives treated and untreated wastewater discharges from several municipalities. Using both DNA and culture-based bacterial analyses, we found that both treated and untreated sewage influences water and sediment bacterial communities in this tributary, and water-sediment exchanges of enteric bacteria and genetic material. Our results also indicated that the treated sewage effluent on this waterway serves as a concentrated source of intI1 (antibiotic resistance) genes, which appear to collect in the sediments below the outfall along with fecal indicator bacteria. Our work also captured the environmental impact of a large rain event that perturbed bacterial populations in sediment and water matrices, independently from the outflow. This study suggests that large precipitation events are an important cause of bacterial and DNA contamination for freshwater tributaries, with runoff from the surrounding environment being an important factor.202540096758
7186170.9957Removal of selected sulfonamides and sulfonamide resistance genes from wastewater in full-scale constructed wetlands. Sulfonamides are high-consumption antibiotics that reach the aquatic environment. The threat related to their presence in wastewater and the environment is not only associated with their antibacterial properties, but also with risk of the spread of drug resistance in bacteria. Therefore, the aim of this work was to evaluate the occurrence of eight commonly used sulfonamides, sulfonamide resistance genes (sul1-3) and integrase genes intI1-3 in five full-scale constructed wetlands (CWs) differing in design (including hybrid systems) and in the source of wastewater (agricultural drainage, domestic sewage/surface runoff, and animal runs runoff in a zoo). The CWs were located in low-urbanized areas in Poland and in Czechia. No sulfonamides were detected in the CW treating agricultural tile drainage water. In the other four systems, four sulfonamide compounds were detected. Sulfamethoxazole exhibited the highest concentration in those four CWs and its highest was 12,603.23 ± 1000.66 ng/L in a CW treating a mixture of domestic sewage and surface runoff. Despite the high removal efficiencies of sulfamethoxazole in the tested CWs (86 %-99 %), it was still detected in the treated wastewater. The sul1 genes occurred in all samples of raw and treated wastewater and their abundance did not change significantly after the treatment process and it was, predominantly, at the level 10(5) gene copies numbers/mL. Noteworthy, sul2 genes were only found in the influents, and sul3 were not detected. The sulfonamides can be removed in CWs, but their elimination is not complete. However, hybrid CWs treating sewage were superior in decreasing the relative abundance of genes and the concentration of SMX. CWs may play a role in the dissemination of sulfonamide resistance genes of the sul1 type and other determinants of drug resistance, such as the intI1 gene, in the environment, however, the magnitude of this phenomenon is a matter of further research.202438081427
3516180.9957Presence of microplastic particles increased abundance of pathogens and antimicrobial resistance genes in microbial communities from the Oder river water and sediment. High abundance of microplastic particles (MPs) in the water environment could be a factor in spreading of pathogens and antimicrobial resistance genes (AMR), especially antibiotic resistance genes (ARGs). The aim of our study was to assess changes in the microbial community developing on microplastic surfaces incubated in water from the Oder River-one of Central Europe's major rivers, flowing through three countries (Czechia, Germany, and Poland)-whose diverse, 20,000-km(2) catchment area (encompassing industrial, agricultural, and urban regions) ensures a relatively high abundance of microbial communities.Samples of water and sediment were taken from river in Wroclaw area. Then the water was poured into disinfected glass liquid containers and pre-drained sediment was added. Control samples of water and sediment were collected on day 0. Then microplastic particles were added (500 mg; ~ 1 mm). Subsequent sampling was performed after incubation on 7(th) and 14(th) day. From each group, samples of sediment and water were collected after the incubation period (n = 5/group), for extraction of microbial DNA and library preparation. Sequencing was performed, using MinION sequencer with 10.4.1 Flow cell. Galaxy Europe platform and R program (v 4.3.3), alpha diversity and PERMANOVA with Benjamini-Hochberg p-value correction for multiple comparisons were used. For identification of biomarker taxa being different between groups, ANCOMBC (Analysis of Compositions of Microbiomes with Bias Correction) was performed. Obtained results shown higher abundance of pathogenic bacteria such as Aeromonas salmonicida Vibrio spp., Escherichia coli or Salmonella after 7 days of incubation in water and sediment. Additionally, after 7 days of incubation numbers of ARGs was higher compared to control group.202540348784
7117190.9957Persistence of antibiotic resistance genes in beef cattle backgrounding environment over two years after cessation of operation. Confined animal feeding operations can facilitate the spread of genes associated with antibiotic resistance. It is not known how cattle removal from beef cattle backgrounding operation affects the persistence of antibiotic resistance genes (ARGs) in the environment. We investigated the effect of cessation of beef cattle backgrounding operation on the persistence and distribution of ARGs in the beef cattle backgrounding environment. The study was conducted at a pasture-feedlot type beef cattle backgrounding operation which consisted of feeding and grazing areas that were separated by a fence with an access gate. Backgrounding occurred for seven years before cattle were removed from the facility. Soil samples (n = 78) from 26 georeferenced locations were collected at the baseline before cattle were removed, and then one year and two years after cattle were removed. Metagenomic DNA was extracted from the soil samples and total bacterial population (16S rRNA), total Enterococcus species and class 1 integrons (intI1), and erythromycin (ermB and ermF), sulfonamide (sul1 and sul2) and tetracycline (tetO, tetW and tetQ) resistance genes were quantified. Concentrations of total bacteria, Enterococcus spp., class 1 integrons, and ARGs were higher in the feeding area and its immediate vicinity (around the fence and the gate) followed by a gradient decline along the grazing area. Although the concentrations of total bacteria, Enterococcus spp., class 1 integrons and ARGs in the feeding area significantly decreased two years after cattle removal, their concentrations were still higher than that observed in the grazing area. Higher concentrations over two years in the feeding area when compared to the grazing area suggest a lasting effect of confined beef cattle production system on the persistence of bacteria and ARGs in the soil.201930768641