Spatial and seasonal distribution of selected antibiotics in surface waters of the Pearl Rivers,China

The distribution and occurrence of 15 antibiotics in surface water of the Pearl River System (Liuxi River, Shijing River and Zhujiang River) and effluents of four wastewater treatment plants (WWTPs) were investigated in two sampling events representing wet season and dry season by using rapid resolution liquid chromatography–electrospray tandem mass spectrometry (RRLC-MS/MS) in positive ionization mode. Only eight antibiotics (sulfadiazine, sulfapyridine, sulfamethazine, sulfamethoxazole, trimethoprim, roxithromycin, erythromycin-H₂O and norfloxacin) were detected in the water samples of the three rivers and the effluents. The detection frequencies and levels of antibiotics in the dry season were higher than those in the wet season. This could be attributed to the dilution effects in the wet season and relatively lower temperature in the dry season under which antibiotics could persist for a longer period. The levels of the detected antibiotics in different sites are generally in a decreasing order as follows: Shijing River ?WWTP effluent ?Zhujiang River ? Liuxi River. Risk assessment based on the calculated risk quotients showed that only erythromycin-H₂O and roxithromycin detected in the Pearl Rivers might have adverse effects on aquatic organisms.     

抗生素最小抑制浓度法(MICs)评估环境大肠杆菌抗生素抗性

2015年世界卫生组织将抗生素的滥用列为21世纪最大的挑战之一,全球范围内抗生素抗药性的散播已严重威胁人类的健康。如何检测环境中的细菌抗药性,并有效评估抗药性感染的风险,是环境微生物研究的一项重要课题。本文通过改进的培养基微量稀释(broth micro-dilution)法,确定了2个地区(中国成都和美国夏威夷)不同环境来源(天然水系和市政污水)的大肠杆菌的抗生素最小抑制浓度(minimum inhibitory concentrations,MICs)。计算所得的MIC分位数(MIC50和MIC90)、菌体抗性百分比及多抗药性指数(multiple antibiotic resistance indexes,MARIs)显示两地不同环境区划的抗生素抗性存在明显的差异。天然水系(成都锦江)中的抗生素抗性是随时间可变的,与当地的降雨事件相关。环境菌株的抗药性模式通过聚类和非度量多维测度(non-metric multidimensional scaling,NMDS)进行分析。广谱β-内酰胺酶基因筛查显示出抗性基因与抗性表型之间的正相关性。结合现有的两地抗生素的使用数据讨论了两地环境抗生素抗性与当地人类活动及抗生素的使用实践之间的紧密联系。采集环境菌株抗生素MIC数据的实验及数据分析方法实现了环境抗药性的跨时空对比,为规范抗生素的区域性使用提供了指导作用。     

Occurrence and fate of typical antibiotics in wastewater treatment plants in Harbin,North-east China

• The concentration distributions and removals of 12 antibiotics were investigated. • Macrolides and fluoroquinolones presented relatively higher concentrations. • The removal of antibiotics did not differ among the different treatment processes. As a new pollutant, antibiotics in the environment and their removal in wastewater treatment plants (WWTPs) have gained considerable attention. However, few studies investigated antibiotics in the north-eastern areas of China. By employing high-performance liquid chromatography-tandem mass spectrometry, the concentration distribution and removal of 12 antibiotics belonging to four types (cephalosporins, sulfonamides, fluoroquinolones, and macrolides) were investigated in influents and effluents as well as the removal efficiencies of four typical treatment processes in 18 wastewater treatment plants of Harbin City, north-eastern China. Macrolides and fluoroquinolones presented relatively higher detection concentrations and rates both in the water and in the sludge phases. Sulfonamides and cephalosporins displayed higher detection concentrations in the water phase. The representative antibiotics in influents and effluents included ROX, NOR, OFL, SMX, AZI, and CLA, with detection rates above 95%. The detection rates of the other six antibiotics exceeded 30%. In the sludge, the other five antibiotics, except SMX, presented relatively higher detection concentrations and rates, with detection rates greater than 95%. For cephalosporin, the detection rates of CTX and CFM in the sludge were 0. The removal efficiencies for the 12 antibiotics differed greatly, with average values ranging from 30% to 80% and the highest values for cephalosporin. The removal of antibiotics did not differ significantly among the four different treatment processes.     

Residual veterinary antibiotics in swine manure from concentrated animal feeding operations in Shandong Province, China

The scientific interest in the occurrence and fate of antibiotics in animal husbandry has increased during the past decades because of the emergence and development of antimicrobial resistance in pathogenic bacteria. This study developed a method for simultaneous detection of five sulfonamides, three tetracyclines and one macrolide in swine manure with stable recoveries (73.0-110.6%) and high sensitivity (limit of quantification <90 μg kg⁻¹). Thereafter, a total of 126 swine manure samples, collected from 21 concentrated animal feeding operations (CAFOs) in Shandong Province of China during summer and winter, were analyzed. The potential influences of different sampling seasons, swine types and food sources on residual antibiotic concentrations were examined in detail. The maximum concentration of residual antibiotic could reach up to 764.4 mg kg⁻¹ (chlortetracycline), and the detection frequencies were 84.9-96.8% for tetracyclines, 0.8-51.6% for sulfonamides and 4.8% for macrolide. These data reveal that antibiotics were extensively used in CAFOs in this district and the manure may act as a non-specific source of antibiotic residue in farmlands and aquatic environments.     

Occurrence, distribution and seasonal variation of antibiotics in the Huangpu River, Shanghai, China

Water samples were collected from 19 sampling sites along the Huangpu River in June and December 2009. The occurrence, distribution and seasonal variation of 22 antibiotics, including four tetracyclines, three chloramphenicols, two macrolides, six fluoroquinolones, six sulfonamides and trimethoprim were investigated. It was found that all 19 sampling sites were contaminated by antibiotics. Four antibiotics (sulfamerazine, norfloxacin, fleroxacin and sarafloxacin) were not detected. The detection frequencies of the other 18 antibiotics were in the range of 5.3-100%. The median concentrations of the detected antibiotics ranged from quantification limits to 36.71 ng L⁻¹ (tetracycline) in June and to 313.44 ng L⁻¹ (sulfamethazine) in December. The number of detected antibiotics and the overall antibiotic concentrations were higher in December than in June due to the different river flow conditions. Different dominant antibiotics were observed for each group of antibiotics between June and December. Higher total concentrations of veterinary antibiotics such as tetracyclines were observed in suburban sampling sites than in unban sites, indicating the role of livestock and agricultural activities as an important source of antibiotic contamination.     

Occurrence of antibiotics in water, sediments, aquatic plants, and animals from Baiyangdian Lake in North China

This study investigated the presence and distribution of 22 antibiotics, including eight quinolones, nine sulfonamides and five macrolides, in the water, sediments, and biota samples from Baiyangdian Lake, China. A total of 132 samples were collected in 2008 and 2010, and laboratory analyses revealed that antibiotics were widely distributed in the lake. Sulfonamides were the dominant antibiotics in the water (0.86-1563 ng L⁻¹), while quinolones were prominent in sediments (65.5-1166 μg kg⁻¹) and aquatic plants (8.37-6532 μg kg⁻¹). Quinolones (17.8-167 μg kg⁻¹) and macrolides [from below detection limit (BDL) to 182 μg kg⁻¹] were often found in aquatic animals and birds. Salvinia natans exhibited the highest bioaccumulation capability for quinolones among three species of aquatic plants. Geographical differences of antibiotic concentrations were greatly due to anthropogenic activities. Sewage discharged from Baoding City was likely the main source of antibiotics in the lake. Risk assessment of antibiotics on aquatic organisms suggested that algae and aquatic plants might be at risk in surface water, while animals were likely not at risk.     

Occurrence, spatial distribution and ecological risks of antibiotics in soil in urban agglomeration

Antibiotics in soil environment are regarded as emerging pollutants and have introduced increasing risks to soil ecosystem and human health in rapid urbanization areas. Identifying the occurrence and spatial variability of antibiotics in soils is an urgent issue in sustaining soil security. In this study, antibiotics in soils were investigated and analyzed in Beijing-Tianjin-Hebei urban agglomeration. The occurrence, spatial distribution, and related affecting factors of antibiotics in soils were identified and ecological risks of antibiotics in soil environment were assessed. Results showed that (1) The mean concentration of soil antibiotics in Beijing-Tianjin-Hebei urban agglomeration was 21.79 µg/kg. Land use substantially affected the occurrence and concentration of antibiotics in soils. Concentrations of antibiotics in cropland and orchard soils were 2-3 times higher than the other land use types. (2) The concentrations of antibiotics in soils in Beijing-Tianjin-Hebei urban agglomeration presented a spatial pattern of high values in southeast, and low values in northwest. Spatial variability of antibiotics in soils was closely related to the application of organic fertilizer and wastewater irrigation as well as topographical features. Furthermore, soil properties and land management policy had substantial influences on soil antibiotics, and soil heavy metals may aggravate the accumulation of antibiotics in soils. (3) Ecological risks assessment of antibiotics in soils demonstrated that erythromycin (ERY), sulfamethoxazole (SMX), and doxycycline (DOX) may introduce high risks to soil ecosystem health, and more attention should be paid to the areas with intensive human activities that had potential high risk to soil ecosystem health. This study suggests that scientific land and soil management should be considered to prevent soil antibiotic pollution and sustain soil security in urban agglomeration.     

Stimulants and donors promote megaplasmid pND6-2 horizontal gene transfer in activated sludge

The activated sludge process is characterized by high microbial density and diversity, both of which facilitate antibiotic resistance gene transfer. Many studies have suggested that antibiotic and non-antibiotic drugs at sub-inhibitory concentrations are major inducers of conjugative gene transfer. The self-transmissible plasmid pND6-2 is one of the endogenous plasmids harbored in Pseudomonas putida ND6, which can trigger the transfer of another co-occurring naphthalene-degrading plasmid pND6-1. Therefore, to illustrate the potential influence of stimulants on conjugative transfer of pND6-2, we evaluated the effects of four antibiotics (ampicillin, gentamycin, kanamycin, and tetracycline) and naphthalene, on the conjugal transfer efficiency of pND6-2 by filter-mating experiment. Our findings demonstrated that all stimulants within an optimal dose promoted conjugative transfer of pND6-2 from Pseudomonas putida GKND6 to P. putida KT2440, with tetracycline being the most effective (100 µg/L and 10 µg/L), as it enhanced pND6-2-mediated intra-genera transfer by approximately one hundred-fold. Subsequently, seven AS reactors were constructed with the addition of donors and different stimulants to further elucidate the conjugative behavior of pND6-2 in natural environment. The stimulants positively affected the conjugal process of pND6-2, while donors reshaped the host abundance in the sludge. This was likely because stimulant addition enhanced the expression levels of conjugation transfer-related genes. Furthermore, Blastocatella and Chitinimonas were identified as the potential receptors of plasmid pND6-2, which was not affected by donor types. These findings demonstrate the positive role of sub-inhibitory stimulant treatment on pND6-2 conjugal transfer and the function of donors in re-shaping the host spectrum of pND6-2.     

Environmental fate of tetracycline resistance genes originating from swine feedlots in river water

Tetracyclines are antibiotics commonly used in swine farms to treat disease and promote growth. However, there are growing concerns regarding the discharge of animal feces into the environment owing to the potential for development and dissemination of tetracycline resistance genes (TRGs). In this study, farming wastewater from one Chinese swine farm as well as river water from seven locations downstream of the farm was sampled. Polymerase chain reaction (PCR) showed that 12 TRGs, including six efflux pump genes (tet(B), tet(C), tet(D), tet(E), tet(G) and tet(L)), five ribosomal protection proteins (RPPs) genes (tet(O), tet(M), tet(Q), tet(W) and tet(S)), and one enzymatic modification gene (tet(X)), were present in all wastewater and river water samples. Quantitative real-time PCR (qPCR) showed that the abundance of tet(C), tet(X), tet(O), tet(M), tet(Q) and tet(W) decreased with downstream flow. Among the detected TRGs, tet(C) had the highest abundance, ranging from 459.5 copies/16S rRNA gene copies in wastewater to 33.8 copies/16S rRNA gene copies in river water samples collected from the last location. Furthermore, pig-specific Bacteroidales 16S rRNA genetic marker was quantified by qPCR to determine the level of fecal pollution in the river water. Bivariate correlation analysis confirmed that the total relative abundance of the six TRGs was significantly correlated with the level of swine feces in the aquatic environment (R² = 0.63, P < 0.05), suggesting that swine feces mainly contributed to the spread of TRGs in the river water.

Supplemental materials are available for this article. Go to the publisher's online edition of Journal of Environmental Science and Health, Part A to view the free supplemental file.     

Degradation of sulfonamides during anaerobic composting of swine manure

Residual antibiotics in manure pose a potential threat to public and ecological health as a result of the application of manure from animals treated with antibiotics to land. The environmental fate of sulfonamides (SAs) in swine manure after composting and field application remains largely unknown. We studied the degradation of the antibiotics sulfadiazine (SD), sulfathiazole (ST), and sulfamethazine (SM2) during anaerobic composting. We tested the effects of temperature and antibiotic concentration on degradation rates. We also evaluated the changes in pH, moisture, and biological degradation material in manure spiked with SAs and in a control. Results showed that the 3 SAs decreased by between 52.31% and 90.30% in all 9 treatments following 14 days of anaerobic composting, and the highest removal efficiencies were observed at a temperature of 35°C and initial concentrations of 6.03, 6.48, and 6.32?µg/g of SD, ST, and SM2, respectively, which were degraded by 90.30%, 85.78%, and 75.18%. Removal efficiencies for all SAs correlated well with moisture and biological degradation material of the manure. These results indicate that composting may be a practical and effective way to reduce concentrations of these three SAs in swine manure prior to its land application.