Concentrations Of Antibiotic Resistance Genes Research Articles

RETRACTED: The behavior of antibiotic resistance genes and their associations with bacterial community during poultry manure composting

In this paper, the influence of clay amendment on antibiotic resistance genes (ARGs) fate and their relation with bacterial diversity in poultry manure (PM) composting was evaluated. The results confirmed that 4% clay amendment was significantly reduce the relative abundances (RAs) 94% of ARGs, while higher dosage of clay blended treatments (T5 and T6) composts were observed relatively very high percentage of ARGs as compared to other clay blended treatments. Clay addition play potential role on the occurrence and distribution of ARGs during the composting by altering the overall RAs of Firmicutes and Proteobacteria, and there host bacterial species. The heat map correlation figure also indicated that reduction of ARGs was significantly associated with the alteration of microbial dynamics and physicochemical parameters during the composting. Therefore, the addition of 4% clay in to PM composting system could efficiently mitigate the greater concentration of ARGs in PM compost and also environmental risk associated with its application.

Application of metagenomic analysis for detection of the reduction in the antibiotic resistance genes (ARGs) by the addition of clay during poultry manure composting

The aim of the study was to reduce relative abundance of antibiotic resistant genes (ARGs) in the poultry manure (PM) compost by adding clay and to determine the mechanism of this effect. Five doses of clay additive [at 0% (T1), 2% (T2), 4% (T3), 6% (T4), 8% (T5) and 10% (T6) based on PM dry weight] were compared to explore the mechanism of reduction in ARGs in the PM compost by the addition of clay. The results confirmed that in the initial raw PM from the breeding farm, the ARG concentrations were 1.7–3.01 times higher than that in the domestic PM. High doses of the clay additive play an important role in reduction in the ARGs and are the main factor responsible for significant variations in ARG abundance between the treatment groups. Therefore, we recommend adding high doses of clay (HDC) as an effective means to reduce the maximum percentage of ARGs in PM compost. A heat map correlation study confirmed that HDC addition during the composting process reduced the bioavailable fractions of toxic metals originating from the chicken feed and significantly impacted ARG dispersion.

Profiling of intracellular and extracellular antibiotic resistance genes in tap water.

Antibiotic resistance genes (ARGs) have gained global attention due to their public health threat. Extracelluar ARGs (eARGs) can result in the dissemination of antibiotic resistance via free-living ARGs in natural environments, where they promote ARB transmission in drinking water distribution systems. However, eARG pollution in tap water has not been well researched. In this study, concentrations of eARGs and intracellular ARGs (iARGs) in tap water, sampled at Tianjin, China, were investigated for one year. Fourteen eARG types were found at the highest concentration of 1.3 × 105 gene copies (GC)/L. TetC was detected in 66.7% of samples, followed by sul1, sul2, and qnrA with the same detection frequency of 41.7%. Fifteen iARGs (including tetA, tetB, tetM, tetQ, tetX, sul1, sul2, sul3, ermB, blaTEM, and qnrA) were continuously detected in all collected tap water samples with sul1 and sul2 the most abundant. Additionally, both eARG and iARG concentrations in tap water presented a seasonal pattern with most abundant prevalence in summer. The concentration of observed intracellular sulfonamide resistance genes showed a significantly positive correlation with total nitrogen concentrations. This study suggested that eARG and iARG pollution of drinking water systems pose a potential risk to human public health.

Seasonal Changes in Antibiotic Resistance Genes in Rivers and Reservoirs in South Korea.

The fate of antibiotic resistance genes (ARGs) in aquatic environments, especially in rivers and reservoirs, is receiving growing attention in South Korea because reservoirs are an important source of drinking water in this country. Seasonal changes in the abundance of 11 ARGs and a mobile genetic element () in two reservoirs in South Korea, located near drinking water treatment plants in Cheonan and Cheongju cities, were monitored for 6 mo. In these drinking water sources, total ARG concentrations reached 2.5 × 10 copies mL, which is one order of magnitude higher than in influents of some wastewater treatment plants in South Korea. During the sampling periods in August, October, and November 2016 and January 2017, sulfonamides (), β-lactam antibiotics (), and tetracycline () resistance genes were the most abundant genes at the two sites. The ARG abundance consistently increased in January relative to 16S ribosomal ribonucleic acid (rRNA) counts. General stress responses to oxidative stress and other environmental factors associated with the cold season could be significant drivers of ARG horizontal gene transfer in the environment. Accordingly, removal of ARGs as a key step in water treatment warrants more attention.

Sources of Antibiotic Resistance Genes in a Rural River System.

The increasing prevalence of antibiotic resistance genes (ARGs) in the environment is problematic due to the risk of horizontal gene transfer and development of antibiotic resistant pathogenic bacteria. Using a suite of monitoring tools, this study aimed to investigate the sources of ARGs in a rural river system in Nova Scotia, Canada. The monitoring program specifically focused on the relative contribution of ARGs from a single tertiary-level wastewater treatment plant (WWTP) in comparison to contributions from the upgradient rural, sparsely developed, watershed. The overall gene concentration significantly ( < 0.05) increased downstream from the WWTP, suggesting that tertiary-level treatment still contributes ARGs to the environment. As a general trend, ARG concentrations upstream were found to decrease as proximity to human-impacted areas decreased; however, many ARGs remained above detection limits in headwater river samples, which suggested their ubiquitous presence in this watershed in the absence of obvious pollution sources. Significant correlations with ARGs were found for human fecal marker, and some antibiotics, suggesting that these markers may be useful for prediction and understanding of ARG levels and sources in rural rivers.

Persistence and spatial variation of antibiotic resistance genes and bacterial populations change in reared shrimp in South China

More attention has been paid to the abundance and diversity of antibiotic resistance genes (ARGs) in aquatic environments. However, few studies have investigated the persistence and spatial variation of ARGs in aquatic organisms. This study investigated the occurrence and abundance of ARGs and the bacterial populations in shrimp intestinal tracts during the rearing period in different regions of Guangdong, South China. The results showed that sul1, sul2, qnrD, and floR were the predominant ARGs. Compared with those of juvenile shrimp, the total concentrations of ARGs in the intestinal tract of adult shrimp in three shrimp farms were 2.45–3.92 times higher (p < 0.05), and the bacterial populations in the adult shrimp intestinal tract changed considerably. Redundancy analysis (RDA) showed that the abundance of Proteobacteria, Firmicutes, and Verrucomicrobia in Farms A, B, and C, respectively, were strongly positively correlated with the most abundant and predominant genes (sul1 and qnrD for Farm A; floR and sul2 for Farm B; floR and sul2 for Farm C) in the shrimp intestinal tract. The results of this study indicated that ARGs gained persistence in the developmental stages of the reared shrimp. Different phyla of predominant bacteria were responsible for the increase of ARGs abundance in the shrimp intestinal tract in different regions. This study represents a case study of the persistence and spatial variation of ARGs in aquaculture and can be a reference for the determination of harmful impacts of ARGs on food safety and human health.

Occurrence and distribution of antibiotic resistance genes in the sediments of drinking water sources, urban rivers, and coastal areas in Zhuhai, China.

Antibiotic resistance genes (ARGs) are regarded as emerging contaminants related with human activities. Aquatic environments of an urban city are apt for the persistence and prevalence of ARGs. In this study, we investigated the occurrence and distribution of ARGs and integrase genes in the sediment samples collected from drinking water sources, urban rivers, and coastal areas of Zhuhai, China, in the dry and wet seasons of 2016. The results show that sulfonamide resistance gene of sulII was present at the highest detection frequency (85.71%); and its average concentrations were also the highest in both dry and wet seasons (3.78 × 107 and 9.04 × 107 copies/g sediment, respectively), followed by tetC, tetO, tetA, ermB, dfrA1, and blaPSE-1. Temporally, the concentrations of total ARGs in the wet season were likely higher than those in the dry season; and spatially, the concentrations of total ARGs in the drinking water sources were substantially lower than those in the urban rivers and nearby coastal areas, indicating the different degrees of anthropogenic impact and consequent health risks. Positive correlations were found between intI1 and each quantitative ARG in all wet season samples rather than dry season samples, which suggested higher temperature and more rain in summer might have positive influences on ARG dissemination, especially that mediated by intI1 gene and class I integrons.

Occurrence of antibiotic resistance genes and bacterial pathogens in water and sediment in urban recreational water

The emergence and prevalence of antibiotic resistance genes (ARGs) and pathogens in the environment are serious global health concern. However, information about the occurrence of ARGs and pathogens in recreational water is still limited. Accordingly, we investigated the occurrence of six ARGs and human pathogens in three recreational lakes, and the correlations between ARGs and one mobile genetic element (intI1) were analyzed. The quantitative PCR results showed that the concentration of ARGs ranged from 4.58 × 100 to 5.0 × 105 copies/mL in water and from 5.78 × 103 to 5.89 × 108 copies/g dry weight (dw) in sediment. Sul1 exhibited the highest level among the five quantifiable ARGs. The concentrations of sul1, bla-TEM, and tetX exhibited significant positive correlations with intI1 (p < 0.05), indicating that intI1 may be involved in their proliferation. The detection frequencies of ARGs ranged from 75%–100%, indicating the prevalence of these risks in this region. The concentration of Escherichia coli, Aeromonas spp., Mycobacterium avium, Pseudomonas aeruginosa, and Salmonella enterica ranged from 103 to 105 copies/100 mL in water and 104–106 copies/g dw in sediment. In total, 25% of the samples harbored all pathogen genes, indicating the prevalence of these pathogens in recreational lakes. Furthermore, the next-generation sequencing results showed that 68 genera of pathogens were present, among which Aeromonas, Mycobacterium, and Pseudomonas were the dominant ones in this region, posing a considerable potential health risk to public health. Overall, the widespread distribution of ARGs and pathogens underscores the need to better monitor and mitigate their propagation in recreational lakes and the associated risks to human health.

Fate of antibiotic resistance genes in two Arctic tundra wetlands impacted by municipal wastewater

In the Canadian Arctic, it is common practice to discharge municipal wastewater into tundra wetlands. Antibiotic resistant bacteria and the antibiotic resistance genes (ARGs) they contain can be present in municipal wastewater and there is a scarcity of knowledge on ARGs in wastewater in Arctic environments. This study was initiated on the fate of ARGs in tundra wetland ecosystems impacted by anthropogenic wastewater sources in Arctic communities. In the summer season of 2016, two wetlands were studied in the Inuit communities of Sanikiluaq and Naujaat in Nunavut, Canada. Genomic DNA was extracted from both soil and water during the spring freshet and late summer in the wetlands, and a suite of nine clinically relevant ARGs (sul1, sul2, mecA, vanA, qnrS, ermB, tetO, blaTEM, blaCTX-M), and an integron gene (int1) were analyzed using quantitative polymerase chain reaction (qPCR). Hydrological and water quality measurements were conducted in conjunction with the microbiological sampling. Gene targets were consistently present in the wastewater, and throughout both wetlands, except for vanA and mecA. Concentrations of ARGs were greater during the spring freshet, due to short hydraulic retention times (<2 days), which coincided with decreased treatment performance. The environmental resistome in un-impacted wetlands had above detection limit concentrations of int1, sul1, sul2, blaCTX-M in water in Naujaat, and sul1, qnrS and tetO in soil in Sanikiluaq. First-order rate constants were widely variable and specific to the gene target. ARGs were present in concentrations elevated above baseline reference sites in tundra wetlands influenced by municipal wastewater, and hydrological conditions had a large impact on their spatial distribution and levels.

Removal of antibiotic resistance genes in two tertiary level municipal wastewater treatment plants

Raw wastewater can contain high levels of antibiotic resistance genes (ARGs), making municipal wastewater treatment plants (WWTPs) critical for the control of the release of ARGs into the environment. The objective of this study was to investigate how individual treatment steps in two tertiary WWTPs affected the removal (copies/mL) and relative abundance of ARGs (copies/copies 16S rRNA genes). Nine ARG markers, representing resistance to commonly used antibiotics, as well as one integron gene (intl1) to assess ARG mobility potential, were quantified using quantitative real-time PCR (qPCR). Both WWTPs met provincial effluent regulations for removal of carbonaceous oxygen demand (CBOD5) and total suspended solids. Eight of the ten ARG markers (intl1, sul1, sul2, tet(O), ermB, blaCTX-M, blaTEM, qnrS) were detected in all samples. In contrast, mecA was detected intermittently and vanA remained below the detection limit in all samples. The total ARG marker abundances decreased by log 1.77 (p < 0.05) in the plant using an aerated lagoon (AL), and by 2.69 logs (p < 0.05) through treatment in the plant employing a biological nutrient removal (BNR) system. The BNR and secondary clarifier steps in both plants afforded the most removal of ARGs. The relative abundance of ARGs remained unchanged at the AL plant and showed a decreasing trend at the BNR plant. Levels of CBOD5, nitrate and the human Bacteroides fecal marker correlated with ARG concentrations, suggesting these variables may be useful in predicting ARG removal. In conclusion, the effluent coming from the WWTPs contained eight of the studied ARG markers in concentrations ranging from 0.01 to 3.6 log copies/mL, indicating their release into the environment, however, the relative abundance of ARGs was not enriched during treatment in the two WWTPs.

Precipitation influences pathogenic bacteria and antibiotic resistance gene abundance in storm drain outfalls in coastal sub-tropical waters

Stormwater contamination can threaten the health of aquatic ecosystems and human exposed to runoff via nutrient and pathogen influxes. In this study, the concentrations of 11 bacterial pathogens and 47 antibiotic resistance genes (ARGs) were determined by using high-throughput microfluidic qPCR (MFQPCR) in several storm drain outfalls (SDOs) during dry and wet weather in Tampa Bay, Florida, USA. Data generated in this study were also compared with the levels of fecal indicator bacteria (FIB) and sewage-associated molecular markers (i.e., Bacteroides HF183 and crAssphage markers) in same SDOs collected in a recent study (Ahmed et al., 2018). Concentration of FIB, sewage-associated markers, bacterial pathogens and many ARGs in water samples were relatively high and SDOs may be potentially hotspots for microbial contamination in Tampa Bay. Mean concentrations of culturable E. coli and Enterococcus spp. were tenfold higher in wet compared to dry weather. The majority of microbiological contaminants followed this trend. E. coli eaeA, encoding the virulence factor intimin, was correlated with levels of 20 ARGs, and was more frequently detected in wet weather than dry weather samples. The blaKPC gene associated with carbapenem resistant Enterobacteriaceae and the beta-lactam resistant gene (blaNPS) were only detected in wet weather samples. Frequency of integron genes Intl2 and Intl3 detection increased by 42% in wet weather samples. Culturable E. coli and Enterococcus spp. significantly correlated with 19 of 47 (40%) ARG tested. Sewage-associated markers crAssphage and HF183 significantly correlated (p < 0.05) with the following ARGs: intl1, sul1, tet(M), ampC, mexB, and tet(W). The presence of sewage-associated marker genes along with ARGs associated with sewage suggested that aging sewage infrastructure contributed to contaminant loading in the Bay. Further research should focus on collecting spatial and temporal data on the microbiological contaminants especially viruses in SDOs.

Tracking antibiotic resistome during wastewater treatment using high throughput quantitative PCR

Wastewater treatment plants (WWTPs) contain diverse antibiotic resistance genes (ARGs), and thus are considered as a major pathway for the dissemination of these genes into the environments. However, comprehensive evaluations of ARGs dynamic during wastewater treatment process lack extensive investigations on a broad spectrum of ARGs. Here, we investigated the dynamics of ARGs and bacterial community structures in 114 samples from eleven Chinese WWTPs using high-throughput quantitative PCR and 16S rRNA-based Illumina sequencing analysis. Significant shift of ARGs profiles was observed and wastewater treatment process could significantly reduce the abundance and diversity of ARGs, with the removal of ARGs concentration by 1–2 orders of magnitude. Whereas, a considerable number of ARGs were detected and enriched in effluents compared with influents. In particular, seven ARGs mainly conferring resistance to beta-lactams and aminoglycosides and three mobile genetic elements persisted in all WWTPs samples after wastewater treatment. ARGs profiles varied with wastewater treatment processes, seasons and regions. This study tracked the footprint of ARGs during wastewater treatment process, which would support the assessment on the spread of ARGs from WWTPs and provide data for identifying management options to improve ARG mitigation in WWTPs.

Effect of Manure Application Rate and Rainfall Timing on the Leaching of Antibiotic-Resistant Bacteria and Their Associated Genes

In this study, we investigate the effect of application rate and timing of liquid swine slurry on leaching of antibiotic-resistant bacteria (ARB) and their antibiotic resistance genes (ARGs) through soil columns. Swine slurry was added to laboratory soil columns at rates of 5000 or 30,000 gal ac−1. For both application rates, rainfall was applied at either 1, 7, or 21 days after slurry application. Column effluent and the top centimeter of soil in the columns were sampled post-rainfall for cultivable bacteria and quantitative PCR was used to quantify tetracycline, methicillin, β-lactam, and erythromycin resistance genetic determinants. We also conducted similar experiments using swine lagoon slurry spiked with antibiotic-resistant E. coli and Salmonella. We found that the concentration of ARB and ARG recovered in the column effluent following application of the swine lagoon slurry generally decreased with increasing lag time between slurry application and simulated rainfall, though most of these decreases were not statistically significant. Moreover, no statistically significant differences in CFU or GU concentration in the column leachate were observed between the low and high slurry application rates. In the experiments using swine slurry spiked with E. coli and Salmonella, concentrations of both microorganisms eluted from fine sand columns were affected by both slurry application rate and lag time; recovery of ARGs, however, was mostly unaffected, but some differences were observed. In columns packed with loamy sand, no recovery was detected in the column effluent for either organism and recovery of ARG was unaffected by manure application rate or rainfall lag time.

Evaluation of attenuation of pharmaceuticals, toxic potency, and antibiotic resistance genes in constructed wetlands treating wastewater effluents

The performance of constructed wetlands (CWs) in the removal of pharmaceutically active compounds (PhACs) is generally evaluated on the basis of chemical analysis. In this work, we used a combination of chemical, toxicological, and molecular analyses to assess the attenuation of PhACs, toxic potency and antibiotic resistance genes (ARGs) in a field study of three CWs serving as tertiary treatment of wastewater treatment plants. First, 17 PhACs were analysed chemically, of which 14 were detected and seven at concentrations >0.1μg/l. Even though some of the individual PhACs were moderately or highly removed in the CWs investigated, median removal of overall PhACs was approximately 50% in the vertical subsurface flow CW (VSF-CW) with a lower hydraulic loading rate while the removal in the other two free water surface flow CWs (SF-CWs) was negligible. Second, toxic potency of wastewater extracts was assessed in a range of bioassays. Estrogenicity was overall attenuated in CWs, while the neurotoxic potency of wastewater extracts did not decrease after passage through the two CWs investigated. Third, the VSF-CW and one of the SF-CW showed a positive removal of an integrase gene and three ARGs tested. The increased concentrations of ARGs in the other SF-CW, as well as the increase of total bacteria in all CWs, may relate to regrowth of resistance-carrying bacteria. Finally, multivariate analysis shows that most PhACs are positively correlated to the observed toxic potency. Additionally, low removal of organics and nutrients seems to parallel with low removal of PhACs. ARGs positively correlated with organics, nutrients and some PhACs, and the integrase gene but not to the respective antibiotics. The insufficient removal of PhACs, toxic potency, and ARGs indicates the need of an optimal design of CWs as tertiary treatment facilities.

Tetracyclines, sulfonamides and quinolones and their corresponding resistance genes in the Three Gorges Reservoir, China

The Three Gorges Project significantly impacted water quality and ecological balance in this area. The special engineered aquatic environment could be an important reservoir for antibiotic resistance genes (ARGs). Fifteen ARGs corresponding to three groups of antibiotics (tetracyclines, sulfonamides and quinolones) were determined in surface water, soil and sediment in this study. Total concentrations of antibiotics ranged from 21.55 to 536.86ng/L, 3.69 to 438.76ng/g, 15.78 to 213.84ng/g in water, soil and sediment, respectively. Polymerase chain reaction (PCR) of ARGs revealed the presence of two sulfonamide resistance genes (sul1, sul2), five tetracycline resistance genes (tetA, tetB, tetM, tetQ, tetG) and class 1 integron gene (intI1) in all samples. And the relative abundance of sulfonamide resistance genes was generally higher than tetracycline resistance genes in three matrices. Significant correlations (p<0.05) were found between the concentrations of intI1 and ARGs (tetA, tetB, tetM, tetQ, tetG, sul1, sul2), indicating intI1 may facilitate the proliferation and propagation of these genes. Redundancy analysis (RDA) showed distribution of ARGs was related to the certain antibiotics residues, which may exert selective pressure on bacteria and thus enrich the abundance of ARGs. The results of this study could provide useful information for both better understanding and management of the contamination caused by ARGs and related antibiotics in engineered aquatic environments.

Chlorine disinfection increases both intracellular and extracellular antibiotic resistance genes in a full-scale wastewater treatment plant

The emergence and spread of antibiotic resistance has posed a major threat to both human health and environmental ecosystem. Although the disinfection has been proved to be efficient to control the occurrence of pathogens, little effort is dedicated to revealing potential impacts of disinfection on transmission of antibiotic resistance genes (ARGs), particularly for free-living ARGs in final disinfected effluent of urban wastewater treatment plants (UWWTP). Here, we investigated the effects of chlorine disinfection on the occurrence and concentration of both extracellular ARGs (eARGs) and intracellular ARGs (iARGs) in a full-scale UWWTP over a year. We reported that the concentrations of both eARGs and iARGs would be increased by the disinfection with chlorine dioxide (ClO2). Specifically, chlorination preferentially increased the abundances of eARGs against macrolide (ermB), tetracycline (tetA, tetB and tetC), sulfonamide (sul1, sul2 and sul3), β-lactam (ampC), aminoglycosides (aph(2’)-Id), rifampicin (katG) and vancomycin (vanA) up to 3.8 folds. Similarly, the abundances of iARGs were also increased up to 7.8 folds after chlorination. In terms of correlation analyses, the abundance of Escherichia coli before chlorination showed a strong positive correlation with the total eARG concentration, while lower temperature and higher ammonium concentration were assumed to be associated with the concentration of iARGs. This study suggests the chlorine disinfection could increase the abundances of both iARGs and eARGs, thereby posing risk of the dissemination of antibiotic resistance in environments.

Seasonal dynamics of tetracycline resistance gene transport in the Sumas River agricultural watershed of British Columbia, Canada

Environmental transport of contaminants that can influence the development of antibiotic resistance in bacteria is an important concern in the management of ecological and human health risks. Agricultural regions are locales where practices linked to food crop and livestock production can introduce contaminants that could alter the selective pressures for the development of antibiotic resistance in microbiota. This is important in regions where the use of animal manure or municipal biosolids as waste and/or fertilizer could influence selection for antibiotic resistance in pathogenic bacterial species. To investigate the environmental transport of contaminants that could lead to the development of antibiotic resistance in bacteria, a watershed with one of the highest levels of intensity of agricultural activity in Canada was studied; the Sumas River located 60 km east of Vancouver, British Columbia. This two-year assessment monitored four selected tetracycline resistance genes (tet(O), tet(M), tet(Q), tet(W)) and water quality parameters (temperature, specific conductivity, turbidity, suspended solids, nitrate, phosphate and chloride) at eight locations across the watershed. The tetracycline resistance genes (Tcr) abundances in the Sumas River network ranged between 1.47 × 102 and 3.49 × 104 copies/mL and ranged between 2.3 and 6.9 copies/mL in a control stream (located far from agricultural activities) for the duration of the study. Further, Tcr abundances that were detected in the wet season months ranged between 1.3 × 103 and 2.29 × 104 copies/mL compared with dry season months (ranging between 0.6 and 31.2 copies/mL). Highest transport rates between 1.67 × 1011 and 1.16 × 1012 copies/s were observed in November 2005 during periods of high rainfall. The study showed that elevated concentrations of antibiotic resistance genes in the order of 102–104 copies/mL can move through stream networks in an agricultural watershed but seasonal variations strongly influenced specific transport patterns of these genes.

Antibiotic 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.

The occurrence of antibiotic resistance genes in tap water – a review

The study presents a review of the occurrence of genetic determinants of antibiotic resistance in tap water. The aim of this study was also to compare the applied methods for antibiotic resistance genes (ARGs) investigations in tap water. As the concentration of ARGs in treated, drinking water is expected to be very low and may cause problems in a standard isolation procedure, the special emphasis is placed on the applied procedures of DNA extraction and their efficiency. The study presents the first attempts to obtain DNA directly from tap water. Further efforts must be put to determine the final amount of obtained DNA and the presence of chosen ARGs among the molecules.

Impact of point sources on antibiotic resistance genes in the natural environment: a systematic review of the evidence.

There is a growing concern about the role of the environment in the dissemination of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARG). In this systematic review, we summarize evidence for increases of ARG in the natural environment associated with potential sources of ARB and ARG such as agricultural facilities and wastewater treatment plants. A total of 5247 citations were identified, including studies that ascertained both ARG and ARB outcomes. All studies were screened for relevance to the question and methodology. This paper summarizes the evidence only for those studies with ARG outcomes (n = 24). Sixteen studies were at high (n = 3) or at unclear (n = 13) risk of bias in the estimation of source effects due to lack of information or failure to control for confounders. Statistical methods were used in nine studies; three studies assessed the effect of multiple sources using modeling approaches, and none reported effect measures. Most studies reported higher ARG concentration downstream/near the source, but heterogeneous findings hindered making any sound conclusions. To quantify increases of ARG in the environment due to specific point sources, there is a need for studies that emphasize analytic or design control of confounding, and that provide effect measure estimates.