Deciphering the Role of Microbial Extracellular and Intracellular Organic Matter in Antibiotic Photodissipation: Molecular and Fluorescent Profiling under Natural Radiation.

This study addresses existing gaps in understanding the specific involvement of dissolved organic matter (DOM) fractions in antibiotic photolysis, particularly under natural conditions and during DOM photobleaching. Employing fluorescent, chemical, and molecular analysis techniques, it explores the impact of extracellular and intracellular organic matter (EOM and IOM) on the photodissipation of multiclass antibiotics, coupled with DOM photobleaching under natural solar radiation. Key findings underscore the selective photobleaching of DOM fractions, propelled by distinct chemical profiles, influencing DOM-mediated antibiotic photolysis. Notably, lipid-like substances dominate in the IOM, while lignin-like substances prevail in the EOM, each uniquely responding to sunlight and exhibiting selective photobleaching. Sunlight primarily targets fulvic acid-like lignin components in EOM, contrasting the initial changes observed in tryptophan-like lipid substances in IOM. The lower photolability of EOM, attributed to its rich unsaturated compounds, contributes to an enhanced rate of indirect antibiotic photolysis (0.339-1.402 h-1) through reactive intermediates. Conversely, the abundance of aliphatic compounds in IOM, despite it being highly photolabile, exhibits a lower mediation of antibiotic photolysis (0.067-1.111 h-1). The triplet state excited 3DOM* plays a pivotal role in the phototransformation and toxicity decrease of antibiotics, highlighting microbial EOM's essential role as a natural aquatic photosensitizer for water self-purification. These findings enhance our understanding of DOM dynamics in aquatic systems, particularly in mitigating antibiotic risks, and introduce innovative strategies in environmental management and water treatment technologies.

Unraveling the Role of Microalgae in Mitigating Antibiotics and Antibiotic Resistance Genes in Photogranules Treating Antibiotic Wastewater.

Harnessing the potential of specific antibiotic-degrading microalgal strains to optimize microalgal-bacterial granular sludge (MBGS) technology for sustainable antibiotic wastewater treatment and antibiotic resistance genes (ARGs) mitigation is currently limited. This article examined the performance of bacterial granular sludge (BGS) and MBGS (of Haematococcus pluvialis, an antibiotic-degrading microalga) systems in terms of stability, nutrient and antibiotic removal, and fate of ARGs and mobile genetic elements (MGEs) under multiclass antibiotic loads. The systems exhibited excellent performance under none and 50 µg/L mixed antibiotics and a decrease in performance at a higher concentration. The MBGS showed superior potential, higher nutrient removal, 53.9 mg/L/day higher chemical oxygen demand (COD) removal, and 5.2-8.2% improved antibiotic removal, notably for refractory antibiotics, and the system removal capacity was predicted. Metagenomic analysis revealed lower levels of ARGs and MGEs in effluent and biomass of MBGS compared to the BGS bioreactor. Particle association niche and projection pursuit regression models indicated that microalgae in MBGS may limit gene transfers among biomass and effluent, impeding ARG dissemination. Moreover, a discrepancy was found in the bacterial antibiotic-degrading biomarkers of BGS and MBGS systems due to the microalgal effect on the microcommunity. Altogether, these findings deepened our understanding of the microalgae's value in the MBGS system for antibiotic remediation and ARG propagation control.

Chlorination of microcystin-LR in natural water: Kinetics, transformation products, and genotoxicity.

Microcystin-LR (MC-LR), a type of cyanotoxin commonly found in natural water bodies (sources of drinking water), poses a threat to human health due to its high toxicity. It is essential to successfully remove this cyanotoxin from drinking water sources. In this study, chlorine was used to oxidize MC-LR in Milli-Q water (MQ) (control test) and natural water collected from Lake Longhu (LLW) as a drinking water source. The removal efficiency, proposed transformation pathways, and genotoxicity were investigated. In the chlorine dose range investigated (4.0 mg L-1 - 8.0 mg L-1), the apparent second-order rate constants for MC-LR chlorination varied from 21.3 M-1s-1 to 31.9 M-1s-1 in MQ, higher than that in LLW (9.06 M-1s-1 to 17.7 M-1s-1) due to a faster chlorine decay attributed to the water matrix (e.g., natural organic matter) of LLW. Eleven transformation products (TPs) of MC-LR were identified in the two waters. The conjugated diene moieties and benzene ring of Adda moiety (3-amino-9-methoxy-2,6,8-trimethyl-10-phenyldeca-4,6-dienoic acid), and the double bond of Mdha moiety (N-methyldehydroalanine) were the major susceptible reaction sites. Attacking unsaturated bonds by hydroxyl and chlorine radicals to generate monochloro-hydroxy-MC-LR was the primary initial transformation pathway, followed by nucleophilic substitution, dehydration, and cleavage in MC-LR. Chlorine substitution on the benzene ring was also observed. Based on the bacterial reverse-mutation assay (Ames assay), TPs in treated natural water did not induce genotoxicity/mutagenicity. These findings shed light on the role of chlorination in controlling the risk of cyanotoxins in drinking water treatment plants.

Rapid sand filtration for <10 µm-sized microplastic removal in tap water treatment: Efficiency and adsorption mechanisms.

The omnipresence of microplastics (MPs) in potable water has become a major concern due to their potential disruptive effect on human health. Therefore, the effective removal of MPs in drinking water is essential for life preservation. In this study, tap water containing microplastic <10 µm in size was treated using constructed pilot-scale rapid sand filtration (RSF) system to investigate the removal efficiency and the mechanisms involved. The results show that the RSF provides significant capacity for the removal and immobilization of MPs < 10 µm diameter (achieving 98 %). Results showed that silicate sand reacted with MPs through a cooperative assembly process, which mainly involved interception, trapping, entanglement, and adsorption. The MPs were quantified by Flow cytometry instrument. A kinetics study underlined the pivotal role of physio-chemisorption in the removal process. MP particles smaller than absorbents, saturation of adsorbents, and reactor hydrodynamics were identified as limiting factors, which were alleviated by backwashing. Backwashing promoted the desorption of up to 97 % MPs, conducive for adsorbent active site regeneration. These findings revealed the critical role of RSF and the importance of backwashing in removing MPs. Understanding the mechanisms involved in removing microplastics from drinking water is crucial in developing more efficient strategies to eliminate them.

Contrasting effects of phytoplankton aging on microplastic antibiotic adsorption depending on species tolerance, and biofouling level.

Recent studies have reported conflicting results on the effects of biofouling on the adsorption behavior of microplastics (MPs). However, the underlying mechanisms driving the adsorption of MPs undergoing biofouling in aquatic environments remain unclear. This study examined the interactions between polyamide (PA), polyvinyl chloride (PVC) and polyethylene (PE) with two phytoplankton, namely cyanobacteria Microcystis aeruginosa and microalgae Chlorella vulgaris. Results indicated that MP effects on phytoplankton were dose- and crystalline-type dependent, with M. aeruginosa being more sensitive to MP exposure than C. vulgaris in the inhibitory order PA > PE > PVC. Analysis of antibiotic adsorption of the MPs showed significant contributions from CH/π interactions on PE and PVC and hydrogen bonding on PA, which decreased with phytoplankton biofouling and aging. Meanwhile, higher levels of extracellular polymeric substances on microalgae-aged MPs compared to cyanobacteria-aged MPs were conducive to adsorption of antibiotics, mainly through hydrophobic interactions. Overall, promotional and anti-promotional adsorption of antibiotics on MPs was induced by biofouling and aging of microalgae and cyanobacteria, respectively. This study provides deep insights into the specific mechanisms by which biofouling affects MP adsorption in aquatic environments, thus advancing our understanding of this critical environmental issue.

Continuous antibiotic attenuation in algal membrane photobioreactor: Performance and kinetics.

The attenuation of 10 mixed antibiotics along with nutrients in a continuous flow mode by four freshwater microalgae (Haematococcus pluvialis, Selenastrum capricornutum, Scenedesmus quadricauda, and Chlorella vulgaris) was examined in membrane photobioreactors (MPBRs). At lab-scale, consistent removal of both antibiotic and nutrient was shown by H. pluvialis and S. quadricauda, respectively. The system exhibited better performance with enhanced removal at HRT 24 h compared to 12 h and 48 h. The highest removal efficiency of antibiotics was observed in H. pluvialis MPBR, with the mean antibiotic removal values of 53.57%- 96.33%. Biodegradation was the major removal pathway of the antibiotics in the algal-MPBR (AMPBR), while removal by bioadsorption, bioaccumulation, membrane rejection, and abiotic was minor. Then, the bacterial feature was studied and showed significant influence from system hydrodynamics. The kinetics of continuous flow antibiotic removal followed Stover-Kincannon and Grau second-order models, which revealed great potential of AMPBR to withstand antibiotic load. The latter coupled with the computational fluid dynamic simulation was successfully applied for the residual antibiotic prediction and potential system optimization. Overall, these results provide an important reference for continuous flow antibiotic removal using AMPBR.

Induced aging, structural change, and adsorption behavior modifications of microplastics by microalgae.

The effects of microalgal biofouling on microplastic (MP) may differ from bacterial biofouling. In this study, the influence of microalgae on MP surface alteration, structural change, and adsorption of organic micropollutants was evaluated. Virgin polyethylene (PE), polyvinyl chloride (PVC), and polyamide (PA) were each immersed in algal photobioreactor and river freshwater for 30 days to simulate algal and river microbe biofouling respectively. Consequently, their physicochemical changes and adsorption potential of a mixture of six bisphenol analogues (BPA, BPS, BPE, BPB, BPF, BPAF) and two parabens (propyl-paraben, benzyl-paraben) were investigated. Owing to the algal bioactive compounds, major microalgae-induced biofouling and more MP aging than the river microbe aging were observed through fractures, pits, cracks, and algal attachments. Intrusion of algal organic matter and scission of polymeric functional groups were revealed during microalgal immersion and the potential MP aging pathways were proposed. Algal biofouling considerably altered the intrinsic properties of the MPs, consequently the adsorption capacity of PE and PVC was enhanced by 3.04-6.72 and 2.14-8.72 times, respectively. Adsorption process onto algal-aged MPs was pH-dependent, endothermic, non-spontaneous, and favored by hydrogen bonds. Meanwhile, the amide group in PA structure was conducive to organic micropollutant adsorption, which was likely reduced by algal aging and the erosion of the N-H stretching. Moreover, higher adsorption capacities of organic micropollutants were shown by the algal-biofilm PE and PVC than virgin and river microbial biofilm MPs. This study discloses that, biofouling and aging of MPs by microalgae through their bioactive components would engender more incidences on MP properties, organic micropollutants adsorption with associated environmental and health hazards.

Microalgal mediated antibiotic co-metabolism: Kinetics, transformation products and pathways.

The mutual interaction of a microalga Chlorella vulgaris with four antibiotics viz. sulfamethoxazole (SMX), trimethoprim (TMP), azithromycin (AZI), and levofloxacin (LEV) individually and in mixture was studied in batch culture. SMX, TMP, and LEV stimulated algal growth, while AZI inhibited its growth. The Combination Index (CI)-isobologram indicated antagonism of the antibiotic mixture on the growth of C. vulgaris. Higher removal efficiency was observed in the mixed antibiotic than in the single antibiotic batch cultures. Biodegradation was the main antibiotic removal mechanism with a similar antibiotic biosorption pattern in single and mix antibiotic cultures. Scanning electron microscopy and Fourier transform infrared spectrophotometry showed minor biochemical alterations on algal cells surface and a stable algal population. Monod kinetics model was successfully applied to understand the growth with respect to the removal efficiency of C. vulgaris in single and mix antibiotic batch cultures. Results indicated relatively higher specific growth rate in the mix antibiotic batch culture with removal efficiency in the order of SMX > LEV > TMP > AZI. In total, 46 metabolites with 18 novel ones of the four antibiotics were identified by using high-resolution mass spectrometry based on the suspect screening approach to propose the potential transformation pathways. Most of the transformation products demonstrated lower toxicity than their respective parents. These findings implied that C. vulgaris could be an outstanding candidate for advanced treatment of antibiotic removal in wastewater.

Simultaneous analysis of multiclass antibiotic residues in complex environmental matrices by liquid chromatography with tandem quadrupole mass spectrometry.

A simultaneous extraction and cleanup method was optimized and validated for the determination of 40 antibiotics from cephalosporin, fluoroquinolone, lincosamide, macrolide, nitroimidazole, quinolone, sulfonamide and tetracycline groups in sediments by liquid chromatography with tandem quadrupole mass spectrometry (LC-MS/MS). The method involved hydration of freeze-dried sediment sample (2.0 g) with 2.5 ml of 0.1 M Na-EDTA McIlvaine buffer and extraction with 5 ml of MeOH and MeCN (1:3 v/v) followed by dispersive solid phase extraction by using 100 mg mix of C18 and PSA (1:2 w/w) and 50 mg MgSO4 prior to LC-MS/MS analysis. The method was validated for 10, 20, 50 and 100 µg/kg spiking levels by using blank sediment sample obtained from a drinking water reservoir according to the guidelines of European Commission Decision (2002) 2002/657/EC. The method produced clean extracts with generally low matrix effect during LC-MS/MS analysis. The mean recoveries ranged between 24-162%, 48-151%, 51-159%, and 50-149% for 10, 20, 50 and 100 µg/kg spiking levels, respectively, with acceptable precision. The analytical method was sensitive enough to achieve 0.01-34.3 µg/kg and 0.03-115 µg/kg limits of detection and quantitation, respectively. The scope of the method was demonstrated by analyzing complex solid environmental matrices (chicken manure, swine manure, poultry feed and soil) spiked at 10, 20, 50 and 100 µg/kg levels. The method was also applied for the antibiotic analysis in samples with incurred residues. Different matrices in the order of the magnitude as sediments < poultry feed < swine manure < soil < chicken manure were detected with the residues of fluoroquinolone, macrolide, sulfonamide and tetracycline antibiotics.

Dissipation of antibiotics by microalgae: Kinetics, identification of transformation products and pathways.

Dissipation potential of four algae viz. Haematococcus pluvialis, Selenastrum capricornutum, Scenedesmus quadricauda and Chlorella vulgaris was investigated against ten antibiotics (sulfamerazine, sulfamethoxazole, sulfamonomethoxine, trimethoprim, clarithromycin, azithromycin, roxithromycin, lomefloxacin, levofloxacin and flumequine) in a series of synthetic wastewater batch culture experiments, maintained at 20, 50 and 100 µg L-1 initial concentration levels and incubated over a period of 40 days. Generally, the antibiotic removal was achieved with overall dissipation percentage (%) varying among the algal species and different antibiotics. Biodegradation was the major antibiotic removal mechanism from the dissolved fraction, with minor contributions of bioadsorption, bioaccumulation, and abiotic factors. The antibiotics dissipation followed the pseudo-first-order-kinetics with the fastest antibiotic degradation rate achieved by H. pluvialis. The Monod kinetics was successfully applied to explain the relationship between the algal growth and the removal of antibiotics and nutrients in the batch cultures. S. capricornutum and C. vulgaris showed more affinity for the macrolides and fluoroquinolones than sulfonamides, while, H. pluvialis and S. quadiricauda showed relatively higher preference for sulfonamides than the other antibiotic groups. A total of 10 transformation products were identified and the transformation pathway was proposed, accordingly. Most of the transformation products had lower toxicity compared with their parent antibiotics.