N-acetyltransferase 2 acetylator genotype-dependent N-acetylation of 4-aminobiphenyl in cryopreserved human hepatocytes.

Arylamine N-acetyltransferases are xenobiotic-metabolizing enzymes responsible for detoxification of many drugs and carcinogens. Two N-acetyltransferase proteins (NAT1 and NAT2) are expressed in humans and they both N-acetylate aromatic amine carcinogens such as 4-aminobiphenyl. Arylamines such as 4-aminobiphenyl represent a large class of chemical carcinogens. Exposure to 4-aminobiphenyl occurs in the chemical, dye and rubber industries as well as in hair dyes, paints, and cigarette smoke. NAT2 is subject to a genetic polymorphism resulting in rapid, intermediate and slow acetylator phenotypes. We investigated the role of the NAT2 genetic polymorphisms on the N-acetylation of 4-aminobiphenyl in cryopreserved human hepatocytes in which NAT2 genotype and deduced phenotype were determined. Differences in sulfamethazine (selectively N-acetylated via NAT2) and 4-aminobiphenyl (N-acetylated by both NAT1 and NAT2) N-acetylation rates among rapid, intermediate, and slow NAT2 acetylator genotypes were tested for significance by one-way analysis of variance. In vitro 4-aminobiphenyl N-acetyltransferase activities differed significantly between rapid, intermediate and slow acetylators at 10 µM (P = 0.0102) or 100 µM (P = 0.0028). N-acetylation of 4-aminobiphenyl in situ also differed significantly between human hepatocytes from rapid, intermediate, and slow acetylators at 10 µM (P = 0.0015) and 100 µM (P = 0.0216). A gene dose-response relationship was exhibited as intermediate acetylators catalyzed 4-aminobiphenyl N-acetylation both in vitro and in situ at rates arithmetically between rapid and slow acetylators. In conclusion, N-acetylation of 4-aminobiphenyl is NAT2 genotype-dependent in human hepatocytes. These results suggest refinement of the exposure limit and safety for arylamine carcinogens according to NAT2 genotype.

Biodiversity, isolation and genome analysis of sulfamethazine-degrading bacteria using high-throughput analysis.

Sulfamethazine (SM2) is one of the sulfonamide antibiotics that is frequently detected in aquatic environment. Given the complex structure of SM2 and its potential threat to the environment, it is necessary to determine the degradation behavior of high-concentration SM2. The mechanisms of community structure and diversity of activated sludge were analyzed. A novel SM2-degrading strain YL1 was isolated which can degrade SM2 with high concentration of 100 mg L-1. Strain YL1 was identified as Paenarthrobacter ureafaciens and there was also a significant increase in the genus during acclimation. Additional SM2 metabolic mechanisms and genomic information of YL1 were analyzed for further research. The succession of the community structure also investigated the effect of SM2 on the activated sludge. This result not only advances the current understanding of microbial ecology in activated sludge, but also has practical implications for the design and operation of the environmental bioprocesses for treatment of antimicrobial-bearing waste streams.

Biochemical Toxicity and Potential Detoxification Mechanisms in Earthworms Eisenia fetida Exposed to Sulfamethazine and Copper.

The present study investigated the biochemical toxicity and potential detoxification mechanisms in earthworms Eisenia fetida exposed to sulfamethazine (SMZ) (7.5, 15 and 30 mg kg-1) either alone or in combination with Copper (Cu) (100 mg kg-1) in soil. The results showed that increasing concentrations of SMZ in soil activated superoxide dismutase, catalase and glutathione peroxidase isozymes, suggesting reactive oxygen species (ROS) burst in earthworms. Treatment with SMZ and Cu separately or in combination caused protein oxidation and damage, elevating the synthesis of ubiquitin, the 20S proteasome, cytochrome P450 (CYP450), and heat shock protein 70 (HSP70). Such treatments also induced the activities of proteases, endoproteinase (EP) and glutathione S-transferases (GSTs). The results suggested that the ubiquitin-20S proteasome, proteases, EP and HSP70 were involved in degradation or remediation of oxidatively damaged proteins. Elevated levels of CYP450 and GSTs also participated in the detoxification of the earthworms.

The contribution of selected organic substrates to the anaerobic cometabolism of sulfamethazine.

Biodegradation of organic micropollutants is likely to occur due to cometabolism by particular microbial groups. In an effort to identify the stages of anaerobic digestion potentially involved in the biodegradation of the veterinary antimicrobial sulfamethazine (SMZ), the influence of selected carbon sources (sucrose, glucose, fructose, ethanol, meat extract, cellulose, soluble starch, soy oil, acetic acid, propionic acid and butyric acid) on SMZ removal by anaerobic sludge was evaluated in short-term batch experiments. Adsorption to the granular sludge constituted a significant removal mechanism, accounting for 39% of SMZ removal in control experiments. The presence of glucose, fructose, sucrose and meat extract exerted an inducing effect on SMZ degradation, resulting in removal efficiencies of 54, 53, 58 and 61%, respectively, indicating the occurrence of cometabolism. Time courses of sucrose and meat extract degradation revealed markedly distinct organic acid profiles but resulted in similar SMZ removals. Temporal profiles of acetic and propionic acid degradation were not associated with SMZ removal, as changes in SMZ concentration were observed even after the organic acids had been completely removed. The experimental results suggest that SMZ cometabolism is not associated to sucrose hydrolysis, acetoclastic methanogenesis and acetogenesis from propionic acid.

Biofiltration of the antibacterial drug sulfamethazine by the species Chenopodium quinoa and its further biodegradation through anaerobic digestion.

The biofiltering capacity, distribution patterns and degradation of the antimicrobial sulfamethazine (SMT) by halophyte Chenopodium quinoa under hydroponic conditions and its further biodegradation through anaerobic digestion were evaluated. C. quinoa was cultivated for a complete life cycle under different concentrations of SMT (0, 2 and 5mg/L) and sodium chloride (0 and 15g/L). C. quinoa is able to uptake and partially degrade SMT. The higher the SMT concentration in the culture medium, the higher the SMT content in the plant tissue. SMT has different distribution patterns within the plant organs, and no SMT is found in the seeds. Dry crop residues containing SMT have a great potential to produce methane through anaerobic digestion and, in addition, SMT is further biodegraded. The highest specific methane yields are obtained using crop residues of the plants cultivated in the presence of salt and SMT with concentrations between 0 and 2mg/L.

Degradation of sulfadiazine, sulfachloropyridazine and sulfamethazine in aqueous media.

Antibiotics discharged to the environment constitute a main concern for which different treatment alternatives are being studied, some of them based on antibiotics removal or inactivation using by-products with adsorbent capacity, or which can act as catalyst for photo-degradation. But a preliminary step is to determine the general characteristics and magnitude of the degradation process effectively acting on antibiotics. A specific case is that of sulfonamides (SAs), one of the antibiotic groups most widely used in veterinary medicine, and which are considered the most mobile antibiotics, causing that they are frequently detected in both surface- and ground-waters, facilitating their entry in the food chain and causing public health hazards. In this work we investigated abiotic and biotic degradation of three sulfonamides (sulfadiazine -SDZ-, sulfachloropyridazine -SCP-, and sulfamethazine -SMT-) in aqueous media. The results indicated that, in filtered milliQ water and under simulated sunlight, the degradation sequence was: SCP > SDZ ≈ SMT. Furthermore, the rate of degradation clearly increased with the raise of pH: at pH 4.0, half-lives were 1.2, 70.5 and 84.4 h for SCP, SDZ and SMT, respectively, while at pH 7.2 they were 2.3, 9.4 and 13.2 h for SCP, SMT and SDZ. The addition of a culture medium hardly caused any change in degradation rates as compared to experiments performed in milliQ water at the same pH value (7.2), suggesting that in this case sulfonamides degradation rate was not affected by the presence of some chemical elements and compounds, such as sodium, chloride and phosphate. However, the addition of bacterial suspensions extracted from a soil and from poultry manure increased the rate of degradation of these antibiotics. This increase in degradation cannot be attributed to biodegradation, since there was no degradation in the dark during the time of the experiment (72 h). This indicates that photo-degradation constitutes the main removal mechanism for SAs in aqueous media, a mechanism that in this case was favored by humic acids supplied with the extracts from soil and manure. The overall results could contribute to the understanding of the environmental fate of the three sulfonamides studied, aiding to program actions that could favor their inactivation, which is especially relevant since its dissemination can involve serious environmental and public health risks.

Catalytic properties and heat stabilities of novel recombinant human N-acetyltransferase 2 allozymes support existence of genetic heterogeneity within the slow acetylator phenotype.

Human N-acetyltransferase 2 (NAT2) catalyzes the N-acetylation of numerous aromatic amine drugs such as sulfamethazine (SMZ) and hydrazine drugs such as isoniazid (INH). NAT2 also catalyzes the N-acetylation of aromatic amine carcinogens such as 2-aminofluorene and the O- and N,O-acetylation of aromatic amine and heterocyclic amine metabolites. Genetic polymorphism in NAT2 modifies drug efficacy and toxicity as well as cancer risk. Acetyltransferase catalytic activities and heat stability associated with six novel NAT2 haplotypes (NAT2*6C, NAT2*14C, NAT2*14D, NAT2*14E, NAT2*17, and NAT2*18) were compared with that of the reference NAT2*4 haplotype following recombinant expression in Escherichia coli. N-acetyltransferase activities towards SMZ and INH were significantly (p < 0.0001) lower when catalyzed by the novel recombinant human NAT2 allozymes compared to NAT2 4. SMZ and INH N-acetyltransferase activities catalyzed by NAT2 14C and NAT2 14D were significantly lower (p < 0.001) than catalyzed by NAT2 6C and NAT2 14E. N-Acetylation catalyzed by recombinant human NAT2 17 was over several hundred-fold lower than by recombinant NAT2 4 precluding measurement of its kinetic or heat inactivation constants. Similar results were observed for the O-acetylation of N-hydroxy-2-aminofluorene and N-hydroxy-2-amino-1-methyl-6-phenylimidazo [4,5-b] pyridine and the intramolecular N,O-acetylation of N-hydroxy-N-acetyl-2-aminofluorene. The apparent V max of the novel recombinant NAT2 allozymes NAT2 6C, NAT2 14C, NAT2 14D, and NAT2 14E towards AF, 4-aminobiphenyl (ABP), and 3,2'-dimethyl-4-aminobiphenyl (DMABP) were each significantly (p < 0.001) lower while their apparent K m values did not differ significantly (p > 0.05) from recombinant NAT2 4. The apparent V max catalyzed by NAT2 14C and NAT2 14D were significantly lower (p < 0.05) than the apparent V max catalyzed by NAT2 6C and NAT2 14E towards AF, ABP, and DMABP. Heat inactivation rate constants for recombinant human NAT2 14C, 14D, 14E, and 18 were significantly (p < 0.05) higher than NAT2 4. These results provide further evidence of genetic heterogeneity within the NAT2 slow acetylator phenotype.

Biodegradation of sulfamethazine by an isolated thermophile-Geobacillus sp. S-07.

Sulfamethazine (SM2) is an antimicrobial drug that is frequently detected in manure compost, is difficult to degrade at high temperatures and is potentially threatening to the environment. In this study, a thermophilic bacterium was isolated from the activated sludge of an antibiotics pharmaceutical factory; this bacterium has the ability to degrade SM2 at 70 °C, which is higher than the traditional manure composting temperature. The strain S-07 is closely related to Geobacillus thermoleovorans based on its 16S rRNA gene sequence. The optimal conditions for the degradation of SM2 are 70 °C, pH 6.0, 50 rpm rotation speed and 50 mL of culture volume. More than 95% of the SM2 contained in media was removed via co-metabolism within 24 h, which was a much higher percentage than that of the type strain of G. thermoleovorans. The supernatant from the S-07 culture grown in SM2-containing media showed slightly attenuated antibacterial activity. In addition, strain S-07 was able to degrade other sulfonamides, including sulfadiazine, sulfamethoxazole and sulfamerazine. These results imply that strain S-07 might be a new auxiliary bacterial resource for the biodegradation of sulfonamide residue in manure composting.

Uptake and translocation of sulfamethazine by alfalfa grown under hydroponic conditions.

Antibiotics are routinely used in intensive animal agriculture operations collectively known as Concentrated Animal Feed Operations (CAFO) which include dairy, poultry and swine farms. Wastewater generated by CAFOs often contains low levels of antibiotics and is typically managed in an anaerobic lagoon. The objective of this research is to investigate the uptake and fate of aqueous sulfamethazine (SMN) antibiotic by alfalfa (Medicago sativa) grass grown under hydroponic conditions. Uptake studies were conducted using hydroponically grown alfalfa in a commercially available nutrient solution supplemented with 10mg/L of SMN antibiotic. Analysis of alfalfa sap, root zone, middle one-third, and top portion of the foliage showed varying uptake rate and translocation of SMN. The highest average amount of SMN (8.58µg/kg) was detected in the root zone, followed by the top portion (1.89µg/kg), middle one-third (1.30µg/kg), and sap (0.38µg/kg) samples, indicating a clear distribution of SMN within the sampled regions. The ultraviolet (UV) spectra of parent SMN and translocated SMN identified in different parts of the plant present the possibility of metabolization during the uptake process. Uptake of SMN using alfalfa grown under hydroponic conditions has potential as a promising remediation technology for removal of similar antibiotics from wastewater lagoons.

Effect of acetaminophen on sulfamethazine acetylation in male volunteers.

The effect of acetaminophen on sulfamethazine N-acetylation by human N-acetyltrasferase-2 (NAT2) was studied in 19 (n=19) healthy male volunteers in two different phases. In the first phase of the study the volunteers were given an oral dose of sulfamethazine 500 mg alone and blood and urine samples were collected. After the 10-day washout period the same selected volunteers were again administered sulfamethazine 500 mg along with 1000 mg acetaminophen. The acetylation of sulfamethazine by human NAT2 in both phases with and without acetaminophen was determined by HPLC to establish their respective phenotypes. In conclusion obtained statistics of present study revealed that acetaminophen significantly (P<0.0001) decreased sulfamethazine acetylation in plasma of both slow and fast acetylator male volunteers. A highly significant (P<0.0001) decrease in plasma-free and total sulfamethazine concentration was also observed when acetaminophen was co-administered. Urine acetylation status in both phases of the study was found not to be in complete concordance with that of plasma. Acetaminophen significantly (P<0.0001) increased the acetyl, free and total sulfamethazine concentration in urine of both slow and fast acetylators. Urine acetylation analysis has not been found to be a suitable approach for phenotypic studies.

Evaluation of sulfamethazine sorption and biodegradation by anaerobic granular sludge using batch experiments.

This study investigated the removal of the veterinary antimicrobial sulfamethazine (SMZ) using anaerobic granular sludge in batch tests. Adsorption and biodegradation were the main mechanisms involved, with adsorption being properly described by a pseudo-second-order model and a linear adsorption isotherm. The adsorption rate constant ranged from 0.00051 to 0.00587 L µg(-1) h(-1), whereas the SMZ partition coefficient was determined to be 0.0717 L g TVS(-1). Biodegradation depended on the presence of readily available organic matter, indicating the occurrence of cometabolism. The addition of exogenous COD to a 144-h batch run at the concentration level of 100 µg L(-1) increased the efficiency of SMZ removal from 57 to 84%. A two-compartment model was developed and fitted to the experimental results, which established the aqueous phase as the main bioavailable compartment. The results suggested that SMZ conversion in anaerobic reactors benefits from high influent dilution and an exogenous supply of organic matter.

Mechanistic and conformational studies on the interaction of sulfamethazine with human immunoglobulin G by molecular modeling and multi-spectroscopic approach in vitro.

Binding interaction of sulfamethazine (SMZ) with human immunoglobulin G (HIgG) has been explored under physiological conditions. The interaction mechanism was firstly predicted through molecular modeling which showed that several hydrogen bonds participated in stabilizing the SMZ-HIgG complex. Fluorescence spectroscopy, ultraviolet-visible (UV-vis) light absorption and circular dichroism (CD) spectroscopy were used to analyze the binding site, binding constants and effects of SMZ on HIgG stability and secondary structure. The binding parameters and thermodynamic parameters at different temperatures for the reaction have been calculated according to the Scatchard, Sips and Van 't Hoff equations, respectively. Experimental results showed that the quenching mechanism was a static quenching and there was one independent class of binding site on HIgG for SMZ during their interaction. The thermodynamic parameters of the reaction, namely standard enthalpy ΔH(0) and entropy ΔS(0), had been calculated to be -19.12 kJ · mol(-1) and 20.22 J · mol(-1) · K(-1), respectively, which meant that the electrostatic interaction was the predominant intermolecular force in stabilizing the SMZ - HIgG complex. Moreover, the conformational changes of HIgG in the presence of SMZ were confirmed by three-dimensional fluorescence spectroscopy, UV-vis absorption spectroscopy and CD spectroscopy.

Degradation of sulfamethazine by microbial electrolysis cell with nickel-cobalt co-modified biocathode.

In this study, nickel-cobalt co-modified stainless steel mesh (Ni-Co@SSM) was prepared and used as the biocathode in microbial electrolysis cell (MEC) for sulfamethazine (SMT) degradation. The optimal electrochemical performance of the Ni-Co@SSM was obtained at the electrodeposition time of 600 s, electrodeposition current density of 20 mA cm-2, and nickel-cobalt molar ratio of 1:2. The removal of SMT in MEC with the Ni-Co@SSM biocathode (MEC-Ni-Co@SSM) was 82%, which increased by 30% compared with the conventional anaerobic reactor. Thirteen intermediates were identified and the potential degradation pathways of SMT were proposed. Proteobacteria, Firmicutes, Patescibacteria, Chloroflexi, Bacteroidetes, and Euryarchaeota are the dominant bacteria at the phylum level in the MEC-Ni-Co@SSM, which are responsible for SMT metabolism. Due to the electrical stimulation, there was an increase in the abundance of the metabolic function and the genetic information processing. This work provides valuable insight into utilizing MECs for effective treatment of antibiotic-containing wastewater.

Isolation, identification and whole-genome analysis of an Achromobacter strain with a novel sulfamethazine resistance gene and sulfamethazine degradation gene cluster.

A sulfamethazine (SM2) degrading strain, Achromobacter mucicolens JD417, was isolated from sulfonamide-contaminated sludge using gradient acclimation. Optimal SM2 degradation conditions were pH 7, 36 °C, and 5 % inoculum, achieving a theoretical maximum degradation rate of 48 % at 50 ppm SM2. Cell growth followed the Haldane equation across different SM2 concentrations. Whole-genome sequencing of the strain revealed novel functional annotations, including a sulfonamide resistance gene (sul4) encoding dihydropteroate synthase, two flavin-dependent monooxygenase genes (sadA and sadB) crucial for SM2 degradation, and unique genomic islands related to metabolism, pathogenicity, and resistance. Comparative genomics analysis showed good collinearity and homology with other Achromobacter species exhibiting organics resistance or degradation capabilities. This study reveals the novel molecular resistance and degradation mechanisms and genetic evolution of an SM2-degrading strain, providing insights into the bioremediation of sulfonamide-contaminated environments.

Effects of sulfamethazine on microbially-mediated denitrifying anaerobic methane oxidation in estuarine wetlands.

Nitrite/nitrate-dependent anaerobic methane oxidation (n-DAMO) is an important methane (CH4) consumption and nitrogen (N) removal pathway in estuarine and coastal wetlands. Antibiotic contamination is known to affect microbially mediated processes; however, its influences on n-DAMO and the underlying molecular mechanisms remain poorly understood. In the present study, using 13CH4 tracer method combined with molecular techniques, we investigated the responses of n-DAMO microbial abundance, activity, and the associated microbial community composition to sulfamethazine (SMT, a sulfonamide antibiotic, with exposure concentrations of 0.05, 0.5, 5, 20, 50, and 100 µg L-1). Results showed that the effect of SMT exposure on n-DAMO activity was dose-dependent. Exposure to SMT at concentrations of up to 5 µg L-1 inhibited the potential n-DAMO rates (the average rates of nitrite- and nitrate-DAMO decreased by 92.9 % and 79.2 % relative to the control, respectively). In contrast, n-DAMO rates tended to be promoted by SMT when its concentration increased to 20-100 µg L-1 (the average rates of nitrite- and nitrate-DAMO increased by 724.1 % and 630.1 % relative to the low-doses, respectively). Notably, low-doses of SMT suppressed nitrite-DAMO to a greater extent than nitrate-DAMO, indicating that nitrite-DAMO was more sensitive to SMT than nitrate-DAMO. Molecular analyses suggest that the increased n-DAMO activity under high-doses SMT exposure may be driven by changes in microbial communities, especially because of the promotion of methanogens that provide more CH4 to n-DAMO microbes. Moreover, the abundances of n-DAMO microbes at high SMT exposure (20 and 50 µg L-1) were significantly higher than that at low SMT exposure (0.05-5 µg L-1). These results advance our understanding of the ecological effects of SMT on carbon (C) and N interactions in estuarine and coastal wetlands.

Uptake of three sulfonamides from contaminated soil by pakchoi cabbage.

Uptake of three sulfonamides (SAs) including sulfadiazine (SDZ), sulfamethazine (SM2) and sulfamethoxazole (SMZ) by pakchoi cabbage from soil was evaluated by using pot experiment. SDZ, SM2 and SMZ spiked in soil could be taken up by pakchoi cabbage. SM2 and SMZ were accumulated more easily by pakchoi cabbage than SDZ. The dissipation half-lives of SMZ (16.8d) and SM2 (16.7d) in soil were significantly longer than SDZ (10.8d). The higher concentrations of SM2 and SMZ in pakchoi cabbage in comparison with that of SDZ could be attributed to the higher residual concentrations of SM2 and SMZ in soil. Increasing initial concentration of SM2 spiked in soil, the residual concentration of SM2 in soil increased and resulted in promoting SM2 uptake in pakchoi cabbage. Concentrations of SAs in pakchoi cabbage planted in combined sulfonamides polluted soil differ from that in single sulfonamide polluted soil, although the same concentration (5.0mg/kg) of SAs was spiked in soil. Combined SAs pollution in soil may enhance SAs uptake in pakchoi cabbage. The lower bacteria numbers in soil under combined pollution resulted in higher residual SAs concentrations in soil, which could be the main reason for higher SAs concentrations in pakchoi cabbage.

Accelerated Biodegradation of Veterinary Antibiotics in Agricultural Soil following Long-Term Exposure, and Isolation of a Sulfamethazine-degrading sp.

The World Health Organization has identified antibiotic resistance as one of the top three threats to global health. There is concern that the use of antibiotics as growth promoting agents in livestock production contributes to the increasingly problematic development of antibiotic resistance. Many antibiotics are excreted at high rates, and the land application of animal manures represents a significant source of environmental exposure to these agents. To evaluate the long-term effects of antibiotic exposure on soil microbial populations, a series of field plots were established in 1999 that have since received annual applications of a mixture of sulfamethazine (SMZ), tylosin (TYL), and chlortetracycline (CTC). During the first 6 yr (1999-2004) soils were treated at concentrations of 0, 0.01 0.1, and 1.0 mg kg soil, in subsequent years at concentrations of 0, 0.1, 1.0, and 10 mg kg soil. The lower end of this concentration range is within that which would result from an annual application of manure from medicated swine. Following ten annual applications, the fate of the drugs in the soil was evaluated. Residues of SMZ and TYL, but not CTC were removed much more rapidly in soil with a history of exposure to 10 mg/kg drugs than in untreated control soil. Residues of C-SMZ were rapidly and thoroughly mineralized to CO in the historically treated soils, but not in the untreated soil. A SMZ-degrading sp. was isolated from the treated soil. Overall, these results indicate that soil bacteria adapt to long-term exposure to some veterinary antibiotics resulting in sharply reduced persistence. Accelerated biodegradation of antibiotics in matrices exposed to agricultural, wastewater, or pharmaceutical manufacturing effluents would attenuate environmental exposure to antibiotics, and merits investigation in the context of assessing potential risks of antibiotic resistance development in environmental matrices.

Functional fungal pellets self-immobilized by mycelium fragments of Irpex lacteus WRF-IL for efficient degradation of sulfamethazine as the sole carbon source.

In order to achieve an efficient microbial material with dual functions of self-immobilization and sulfamethazine (SMZ) degradation, this study explored the pelletization technique utilizing mycelium fragments of Irpex lacteus WRF-IL and systematically examined the pellets formation conditions and degradation capability. The Box-Behnken design results demonstrated that pure mycelium fragments, broken by frosted glass beads, could be rapidly self-immobilized to form white rot mycelial pellets (WRMPs) within 24 h, serving as the pelleting core. These WRMPs could completely remove SMZ as the sole carbon source within 20 h. The addition of sucrose expedited this process, achieving complete removal within only 14 h. Kinetic analysis showed that WRMPs could potentially remove SMZ at higher concentrations (>25 mg/L). Biodegradation was the primary pathway of SMZ removal. Seven intermediates were identified by QTOF LC/MS, and three transformation pathways initiated by SO2 overflow, molecular rearrangement, and aniline moiety oxidation were deduced.

Immobilization of Bacillus Thuringiensis and applicability in removal of sulfamethazine from soil.

Microbial degradation is considered an essential and promising treatment for sulfadimidine contamination of soil. To address the low colonization rates and inefficiencies of typical antibiotic-degrading bacteria, sulfamethazine (SM2)-degrading strain H38 is converted into immobilized bacteria in this study. Results show that the removal rate of SM2 by immobilized strain H38 reaches 98% at 36 h, whereas the removal rate of SM2 by free bacteria reaches 75.2% at 60 h. In addition, the immobilized bacteria H38 exhibits tolerance to a wide range of pH (5-9) and temperature (20 °C-40 °C). As the amount of inoculation increases and the initial concentration of SM2 decreases, the removal rate of SM2 by the immobilized strain H38 increases gradually. Laboratory soil remediation tests show that the immobilized strain H38 can remove 90.0% of SM2 from the soil on the 12th day, which exceeds the removal by free bacteria by 23.9% in the same period. Additionally, the results show that the immobilized strain H38 enhances the overall activity of microorganisms in SM2-contaminated soil. Compared with the SM2 only (control group containing no bacteria) and free bacterial treatment groups, the gene expression levels of ammonia-oxidizing archaea, ammonia-oxidizing bacteria, cbbLG, and cbbM increased significantly in the treatment group with immobilized strain H38. This study shows that immobilized strain H38 can reduce the effect of SM2 on soil ecology to a greater extent than free bacteria, while providing safe and effective remediation.

Effect of subtherapeutic and therapeutic sulfamethazine concentrations on transcribed genes and translated proteins involved in Microbacterium sp. C448 resistance and degradation.

Microbacterium sp. C448, isolated from a soil regularly exposed to sulfamethazine (SMZ), can use various sulphonamide antibiotics as the sole carbon source for growth. The basis for the regulation of genes encoding the sulphonamide metabolism pathway, the dihydropteroate synthase sulphonamide target (folP), and the sulphonamide resistance (sul1) genes is unknown in this organism. In the present study, the response of the transcriptome and proteome of Microbacterium sp. C448 following exposure to subtherapeutic (33 µM) or therapeutic (832 µM) SMZ concentrations was evaluated. Therapeutic concentration induced the highest sad expression and Sad production, consistent with the activity of SMZ degradation observed in cellulo. Following complete SMZ degradation, Sad production tended to return to the basal level observed prior to SMZ exposure. Transcriptomic and proteomic kinetics were concomitant for the resistance genes and proteins. The abundance of Sul1 protein, 100-fold more abundant than FolP protein, did not change in response to SMZ exposure. Moreover, non-targeted analyses highlighted the increase of a deaminase RidA and a putative sulphate exporter expression and production. These two novel factors involved in the 4-aminophenol metabolite degradation and the export of sulphate residues formed during SMZ degradation, respectively, provided new insights into the Microbacterium sp. C448 SMZ detoxification process.