This study explores antibiotic resistance genes (ARGs) as emerging environmental contaminants. The purpose of this study was to investigate the occurrence of ARGs in various environmental compartments in northern Colorado, including Cache La Poudre (Poudre) River sediments, irrigation ditches, dairy lagoons, and the effluents of wastewater recycling and drinking water treatment plants. Additionally, ARG concentrations in the Poudre River sediments were analyzed at three time points at five sites with varying levels of urban/agricultural impact and compared with two previously published time points. It was expected that ARG concentrations would be significantly higher in environments directly impacted by urban/agricultural activity than in pristine and lesser-impacted environments. Polymerase chain reaction (PCR) detection assays were applied to detect the presence/absence of several tetracycline and sulfonamide ARGs. Quantitative real-time PCR was used to further quantify two tetracycline ARGs (tet(W) and tet(O)) and two sulfonamide ARGs (sul(I) and sul(II)). The following trend was observed with respect to ARG concentrations (normalized to eubacterial 16S rRNA genes): dairy lagoon water > irrigation ditch water > urban/agriculturally impacted river sediments (p < 0.0001), except for sul(II), which was absent in ditch water. It was noted that tet(W) and tet(O) were also present in treated drinking water and recycled wastewater, suggesting that these are potential pathways for the spread of ARGs to and from humans. On the basis of this study, there is a need for environmental scientists and engineers to help address the issue of the spread of ARGs in the environment.

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The occurrence of 15 antibiotics belonging to three different groups, tetracyclines (TCs), sulfonamides (SAs), and macrolides (MLs), mainly used to prevent or treat illness for humans and also to control disease or to promote the growth for animals was studied in aqueous and sediment matrices. The result of spatial and temporal statistical analysis revealed that measured concentrations of individual antibiotics were significantly different depending on sampling location and time periods for aqueous and sediment samples. High concentrations of human-used antibiotics were detected downstream of a wastewater treatment plant, and animal-used antibiotics were mainly found in a region with significant agricultural activity. Generally, the highest concentrations of antibiotics for both water and sediment samples were measured in winter indicating that low flow conditions and cold-water temperatures might enhance the persistence of these compounds. Furthermore, a pseudo-partitioning coefficient(P-PC) was introduced to provide a better understanding of the partitioning of antibiotics into the sediment. Different P-PC values were found depending on the sorption characteristics of the individual antibiotics. Sediment samples showed a greater detection frequency and a much higher concentration compared to aqueous samples taken at the same site. Since microorganism antibiotic resistance can develop in sediments, the importance of analyzing this matrix is underscored.

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An analytical method was developed and tested for four different groups of veterinary antibiotics in both river water and sediment matrices. Solid phase extraction (SPE) was used to enrich and to clean up the aqueous sample. Also, Mcllvaine and ammonium hydroxide buffer solutions were used to extract the compounds from the sediment matrix. High performance liquid chromatography (HPLC) equipped with tandem mass spectrometry (MS/MS) was used to separate and quantify the samples. The range of recoveries (in percent) for tetracyclines (TCs), sulfonamides (SAs), macrolides (MLs), and ionophore polyethers (IPs) in the water matrix were 102.2-124.8, 76.6-124.3, 89.5-114.7, 82.7-117.5 with 1-13 (%) of relative standard deviation respectively with three different concentrations. For sediment, the percent recovery ranges were 32.8-114.8, 62.4-108.9, 53.4-128.4 and 51.3-105.4 for TCs, SAs, MLs and IPs, respectively. The relative standard deviation ranged from 16 - 27 (%) over three different concentrations. The limit of quantification (LOQ) was determined by two different methods and calculated to be in the range of 0.01-0.04 microg/l and 0.3-2.5 microg/kg for TCs, SAs, and MLs in water and sediment, respectively. For IPs, the LOQ was 0.001-0.003 microg/l in river water and 0.4-3.6 microg/kg for sediment. The sediment concentration measured in an agriculture-influenced river was much higher than in the overlying water matrix, indicating a high degree of sediment partitioning for these compounds.

Research has verified the occurrence of veterinary antibiotics in manure, agricultural fields, and surface water bodies, yet little research has evaluated antibiotic runoff from agricultural fields. The objective of this study was to evaluate the potential for agricultural runoff to contribute antibiotics to surface water bodies in a worst-case scenario. Our hypothesis was that there would be significant differences in antibiotic concentrations, partitioning of losses between runoff and sediment, and pseudo-partitioning coefficients (ratio of sediment concentration to runoff concentration) among antibiotics. An antibiotic solution including tetracycline (TC), chlortetracycline (CTC), sulfathiazole (STZ), sulfamethazine (SMZ), erythromycin (ERY), tylosin (TYL), and monensin (MNS) was sprayed on the soil surface 1 h before rainfall simulation (average intensity = 60 mm h(-1) for 1 h). Runoff samples were collected continuously and analyzed for aqueous and sediment antibiotic concentrations. MNS had the highest concentration in runoff, resulting in the highest absolute loss, although the amount of loss associated with sediment transport was <10%. ERY had the highest concentrations in sediment and had a relative loss associated with sediment >50%. TYL also had >50% relative loss associated with sediment, and its pseudo-partitioning coefficient (P-PC) was very high. The tetracyclines (TC and CTC) had very low aqueous concentrations and had the lowest absolute losses. If agricultural runoff is proven to result in development of resistance genes or toxicity to aquatic organisms, then erosion control practices could be used to reduce TC, ERY, and TYL losses leaving agricultural fields. Other methods will be needed to reduce transport of other antibiotics.

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