Role of a typical swine liquid manure treatment plant in reducing elements of antibiotic resistance.

Biological treatment of swine liquid manure may be a favorable environment for the enrichment of bacteria carrying antibiotic resistance genes (ARGs), raising the alert about this public health problem. The present work sought to investigate the performance of a swine wastewater treatment plant (SWWTP), composed of a covered lagoon biodigester (CLB) followed by three facultative ponds, in the removal of usual pollutants, antibiotics, ARGs (blaTEM, ermB, qnrB, sul1, and tetA), and intI1. The SWWTP promoted a 70% of organic matter removal, mainly by the digester unit. The facultative ponds stood out in the solids' retention carried from the anaerobic stage and contributed to ammonia volatilization. The detected antibiotic in the raw wastewater was norfloxacin (< 0.79 to 60.55 µg L-1), and the SWWTP seems to equalize peaks of norfloxacin variation probably due to sludge adsorption. CLB reduced the absolute abundance of ARGs by up to 2.5 log, while the facultative stage does not seem to improve the quality of the final effluent in terms of resistance elements. Considering the relative abundances, the reduction rates of total and ARG-carrying bacteria appear to be similar. Finally, correlation tests also revealed that organic matter and solids control in liquid manure treatment systems could help reduce the spread of ARGs after the waste final disposal.

Removal of cephalexin and erythromycin antibiotics, and their resistance genes, by microalgae-bacteria consortium from wastewater treatment plant secondary effluents.

Antibiotics have become a concern in the aquatic environments owing to the potential development of bacterial resistances. Thus, this study evaluated the removal of cephalexin (CEP) and erythromycin (ERY) from a local wastewater treatment plant (WWTP) effluent, mediated by microalgae-bacteria consortium. Likewise, the removal of correlated antibiotics resistance genes blaTEM and ermB was also assessed. The incubation results showed that the added concentrations of selected antibiotics did not restrain the consortium growth. Moreover, CEP and ERY were almost completely removed after the cultivation period, reaching total removals of 96.54% and 92.38%, respectively. The symbiotic interaction between microalgae and bacteria plays a role in the kinetics removal of CEP and ERY. The abundance of blaTEM and ermB was reduced by 0.56 and 1.75 logs, respectively. Lastly, our results suggest that technology based on natural microalgae-bacteria consortium could be a potential alternative to improve the quality of WWTP effluents.

Low-temperature partitioning extraction followed by liquid chromatography tandem mass spectrometry determination of multiclass antibiotics in solid and soluble wastewater fractions.

An analytical method based on low-temperature partitioning extraction (LTPE) followed by high performance liquid chromatography coupled to triple quadrupole mass spectrometry analysis was developed and validated for the determination of eight multiclass antibiotics in wastewater. The analyzed target antibiotics included one ß-lactam, two sulfonamides, three fluoroquinolones, one macrolide and one diaminopyrimidine. LTPE parameters such as sample pH, volume ratio between sample and extractor solvent, ultra-sonic extraction time, extraction tube material, solvent and volume to reconstitute the sample extracts, were optimized. Additionally, the influence of solids on extraction efficiency was evaluated. Quantification of the target antibiotics was performed by double consecutive injection method, without the use of a labeled compound, in order to correct matrix effects. The whole samples were analyzed, including, liquid and solid fractions of wastewater. The results revealed that the filtration step can underestimate the total antibiotics concentration, particularly to the hydrophobic compounds that have higher affinity for solids, indicating that the suspended wastewater particulate should not be neglected. The method detection limit ranged from 18.54 ng L-1 (trimethoprim) to 78.49 ng L-1 (ciprofloxacin). Intra-day precision of less than 12.3% was achieved. The recoveries values ranged from 13.9% (sulfadiazine) to 48.9% (erythromycin) in influent samples and from 19.1% (sulfadiazine) to 57.2% (ciprofloxacin) in effluent samples. The method was applied to the measurement of antibiotic residues in influent and effluent from wastewater treatment plants. The majority target antibiotics were detected in wastewater samples. Their concentrations ranged from 237 to 9553 ng L-1 in influent and from 212 to 1660 ng L-1 in effluent. This work provides new insights on the applicability of LTPE for antibiotic residues extraction from wastewater. In addition, the performed analysis highlights the importance of measuring total concentrations of analytes in whole sample.

Aqueous chlorination of herbicide metribuzin: Identification and elucidation of "new" disinfection by-products, degradation pathway and toxicity evaluation.

A widely used herbicide, metribuzin, was evaluated for degradation, mineralization and disinfection by-products (DBPs) formation during aqueous chlorination. In addition, to assess the toxicity effects of chlorination on metribuzin solution the following tests were performed: acute toxicity using Artemia salina nauplii; cell viability using MTT assay; estrogenicity using a re-engineered Bioluminescent Yeast Estrogen Screen (BLYES) and a constitutively bioluminescent strain (BLYR); mutagenicity and developmental toxicity using Q(SAR) methodology. Metribuzin at 10 mg·L-1 was degraded by chlorination, achieving 93% of removal at 30 min of reaction. TOC analysis showed that the herbicide does not suffer complete mineralization, even after 24 h of contact with free chlorine. Seventeen DBPs were detected and their structural formulae were elucidated by high resolution mass spectrometry. Toxicity effects for chlorinated solutions increased when compared to the unreacted metribuzin solution. DBPs were more toxic to Artemia salina nauplii, increasing around 20% on nauplii mortality. It was also observed high estrogenicity to human receptors in BLYES assays and mutagenic and developmental toxicant effects to animals and humans in Q(SAR) methodology, suggesting that DBPs are potentially more toxic than the precursor metribuzin. Metribuzin solutions at 10 mg·L-1 showed equivalent 17-ß-estradiol values ranged from 0.061 to 6.71 µg·L-1 after to be chlorinated at different reaction times.

Biodegradation of sulfamethoxazole by microalgae-bacteria consortium in wastewater treatment plant effluents.

Sulfamethoxazole (SMX) has been commonly detected in wastewater treatment plant (WWTP) effluents. SMX and other antibiotics can be considered as environmental contaminants of emerging concern. Due to their toxicity effects and their potential for the development of bacterial resistance their presence in aquatic compartment becomes a threat to human health. This study evaluated the bioremediation of SMX in WWTP effluents using a tertiary treatment composed by microalgae-bacteria consortium under low intensity artificial LED illumination, and also the assessment of sulfonamide resistance gene (sul1). The removal of SMX from WWTP effluents were 54.34 ± 2.35%, in which the microalgae-bacteria consortium improves the removal performance of SMX. The main process of SMX removal can be attributed to the symbiotic biodegradation by bacteria due to the increase of oxygen released by the microalgae photosynthetic process. Therefore, the microalgae-bacteria consortium used in this study, demonstrated to be a promising alternative for bioremediation of SMX, with potential for removal others contaminants from wastewater effluent. However, the residual SMX and the relative abundance of antibiotics resistance genes (ARG) found in this study suggest that SMX contributes to selective pressure for ARG maintenance and proliferation in WWTP effluent. Thus, further studies to removal ARG from WWTP effluent are needed.

Method development for simultaneous determination of polar and nonpolar pesticides in surface water by low-temperature partitioning extraction (LTPE) followed by HPLC-ESI-MS/MS.

During this research, chemometric approaches were applied for optimization of the low-temperature partitioning extraction (LTPE) for the simultaneous analysis of the pesticides: acephate, difenoconazole, fenamidone, fluazifop, fluazinam, methamidophos, and thiamethoxam from surface water samples and determination by high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry. It was used the 23 full factorial and the Doehlert experimental designs. The extraction technique was optimized by evaluating the effects of the three variables: sample pH, ionic strength (addition of Na2HPO4), and organic solvent volume. Considering the interest to find an optimal condition for all analytes simultaneously, the best extraction parameters found were as follows: pH = 5.33, concentration of Na2HPO4 = 0.0088 mol L-1 and organic phase volume = 4.5 mL. The optimized methodology showed LOD and LOQ levels from 0.33 to 8.13 ng L-1 and from 1.09 to 26.84 ng L-1, respectively. The recovery values ranged from 38.37 and 99.83% and the RSD values varied from 2.33 to 18.92%. The method was applied to surface water analysis sampled in areas with intensive agricultural practices in Ouro Branco City, Minas Gerais, Brazil. The difenoconazole was detected in concentrations between 12.53 and 94.76 ng L-1.