Mechanistic approach of tannery wastewater and sulfadiazine mutual toxicity in wheat (Triticum aestivum L.) and mitigation through exogenous application of gallic acid.

Excessive release of chromium (Cr) from the tanning industry and antibiotics from livestock caused severe hazards to humans. Gallic acid (GA 10 mM) alleviated alone/combined SDZ 30 mg kg-1 and TWW 40, 60, and 100% stress in wheat. GA (10 mM) decreased the TSP 12 and 13%, TFAA 8 and 10%, TSS 14 and 16%, RS 18 and 16%, and NRS 11 and 9% in shoots and grains under SDZ + TWW (30 mg kg-1+100%), compared without foliar. GA (10 mM) declined the MDA 20 and 31, EL 13 and 36%, H2O2 17 and 15%, O2•- 10 and 11% in leaves and roots, under combined SDZ + TWW (30 mg kg-1+100%), compared without foliar. GA (10 mM) improved the POD 106 and 30%, SOD 145 and 31%, CAT 78, and 35%, APX 100 and 25% in leaves and roots under combined SDZ + TWW (30 mg kg-1+100%), compared without foliar application. Considerably GA (10 mM) reduced total Cr 18, CrIII 20, and CrVI 50% in roots and shoots 19, 41, and 48%, and grains 15, 27, and 29% respectively, under combined SDZ + TWW (30 mg kg-1+100%) stress, compared without foliar. Overall, GA boosted the wheat growth, physiology, and defence system by inhibiting the combined SDZ + Cr toxicity.

Tetracycline and sulfadiazine toxicity in human liver cells Huh-7.

As typical antibiotics, tetracycline (TC) and sulfadiazine (SDZ) enter the human body through the food chain. Therefore, it is necessary to understand their individual and combined toxicity. In this study, the effects of TC, SDZ, and their mixture on cell viability, cell membrane damage, liver cell damage, and oxidative damage were evaluated in in vitro assays with human liver cells Huh-7. The results showed cytotoxicity of TC, SDZ, and their mixture, which induced oxidative stress and caused membrane and cell damage. The effect of antibiotics on Huh-7 cells increased with increasing concentration, except for lactate dehydrogenase (LDH) activity that commonly showed a threshold concentration response and cell viability, which commonly showed a biphasic trend, suggesting the possibility of hormetic responses where proper doses are included. The toxicity of TC was commonly higher than that of SDZ when applied at the same concentration. These findings shed light on the individual and joint effects of these major antibiotics on liver cells, providing a scientific basis for the evaluation of antibiotic toxicity and associated risks.

The different toxicological effects and removal efficiencies of norfloxacin and sulfadiazine in culturing Arthrospira (Spirulina) platensis.

Norfloxacin (NFX) and sulfadiazine (SDZ) are two widely used antibiotics belonging to fluoroquinolone and sulfonamide groups, respectively, and have become the commonly detected micropollutants in aquatic environments. However, only few works have been conducted to investigate the highly probable inhibition of these antibiotic pollutants to Arthrospira platensis, which is an important species of cyanobacteria that is one of primary producers in aquatic ecosystems and should be remarkably sensitive to environmental pollutants due to its prokaryotic characteristics. Hence, the toxicological effects and removal efficiencies of NFX and SDZ in culturing A. platensis were studied by analyzing the biomass growth, photosynthetic pigments, primary biocomponents, and antibiotics concentration. The corresponding variations of these characteristics showed the higher sensitivity of A. platensis to NFX than to SDZ, indicating the specifically targeted effect of NFX on A. platensis, which could be confirmed in silico by the higher binding affinity of NFX with the critical enzyme. The obtained results illustrated the roles of NFX and SDZ on the growth of A. platensis, thus providing the great support in employing A. platensis to reduce hazards from contaminated water and recover biomass resources.

Response of submerged macrophytes and leaf biofilms to different concentrations of oxytetracycline and sulfadiazine.

Oxytetracycline and sulfadiazine were widely used and they entered the environment through various channels such as domestic sewage, medical wastewater and agricultural wastewater, causing significant ecological risk. To determine the effects of different antibiotic concentrations on submerged macrophytes, Vallisneria natans was exposed to solutions containing different concentrations of oxytetracycline and sulfadiazine (0.1 mg/L、1 mg/L、10 mg/L、50 mg/L). After 20-days exposure, we found that 10 mg/L groups had a significant effect on Vallisneria natans. Under high antibiotic concentrations, the growth of Vallisneria natans was inhibited, chloroplasts were deformed, the chlorophyll content was reduced, and antioxidant enzyme activities, such as superoxide dismutase and glutathione, were increased. There was no significant difference between the control group and groups with low antibiotic concentrations (≤1 mg/L). The N-acyl-l-homoserine lactone concentration tended to increase with increasing antibiotic concentrations. The presence of antibiotics also affected the microbial community structure of biofilms on the submerged macrophytes. For example, the higher the concentration of antibiotics, the higher the proportion of Proteobacteria. These results suggest that high concentrations of oxytetracycline and sulfadiazine can disrupt homeostasis, induce effective Vallisneria natans defense mechanisms and alter biofilms in aquatic ecosystems.

Insights into the mechanism of the interference of sulfadiazine on soil microbial community and function.

The accumulation of sulfonamides in the soil environment possessed the potential to change soil microbial community and function. Metabolomics is capable of providing insights into the carbon metabolic pool and molecular mechanisms associated with external stressors. Here we evaluated alternations in soil bacterial community and soil metabolites profiles under sulfadiazine (SDZ) exposure and proposed a potential mechanism that SDZ accumulation in soil affected soil organic matter (SOM) cycling. Sequencing analysis showed that the relative abundance of bacterial species associated with carbon cycling significantly decreased under high concentrations of SDZ exposure. Untargeted metabolomics analysis showed that 78 metabolites were significantly changed with the presence of SDZ in soil. The combination of functional predictions and pathway analysis both demonstrated that high concentrations of SDZ exposure could cause disturbance in anabolism and catabolism. Moreover, the noticeable decline in the relative content of carbohydrates under high concentrations of SDZ exposure might weaken physical separation and provide more chances for microbes to degrade SOM. The above results provided evidence that SDZ accumulation in soil held the potential to disturb SOM cycling. These findings spread our understanding about the environmental risk of antibiotic in the soil environment beyond the dissemination of antibiotic resistance.

The effect of bio/organic fertilizers on the phytotoxicity of sulfadiazine to Echium amoenum in a calcareous soil.

The fate of antibiotics and their effects on plant growth may be changed by the application of fertilizers. The present study was carried out to investigate the effect of sulfadiazine (SDZ), rice husk compost (RHC), rice husk biochar (RHB), and mycorrhiza (MR) on the growth attributes of Iranian Echium amoenum Fisch & C.A. Mey. A greenhouse experiment as a completely randomized design with six treatments of bio/organic-fertilizers (no bio-fertilizer (NF), RHB, RHC, MR, RHB+MR, and RHC+MR) and three levels of SDZ application (0, 100, and 200 mg kg-1) was performed for 7months with three replicates. Shoot and root SDZ concentrations were determined using high-pressure liquid chromatography-diode array detection (HPLC-DAD) instrumentation. The results revealed that the application of RHC, RHB, and MR had a significant impact on the reduction of the toxicity effects of SDZ on plant properties. The lowest values of growth parameters belonged to the 200 mg kg-1 of SDZ with no bio-fertilizers, while the highest growth parameters were observed in the treatments of RHB+MR, and RHC+MR with no SDZ application. Also, chlorophyll pigments content was affected by used treatments and the lowest rates of chlorophyll a (4.24), chlorophyll b (2.99), and carotenoids (2.88) were related to the 200 mg kg-1 of SDZ with no biofertilizers application. The co-application of bio-fertilizers and SDZ (at both levels of 100 and 200 mg kg-1) decreased SDZ uptake by both shoot and root in comparison with the control. The same results were obtained with macro (NPK) and micro (Fe, Zn, Cu, and Mn) nutrients uptake by the shoot in which the lowest values of nutrients uptake were observed in treatment of 200 mg kg-1 of SDZ with no bio-fertilizers. Furthermore, in the case of the effect of the used treatments on root colonization, the results showed that the lowest value (7.26%) belonged to the 200 mg kg-1 application of SDZ with no bio-fertilizers. Generally, this study demonstrated that bio-fertilizers could be considered as an effective strategy in controlling the negative effects of antibiotics on the growth properties and nutrients status of the plants grown in such contaminated soils.

The Toxicity Exerted by the Antibiotic Sulfadiazine on the Growth of Soil Bacterial Communities May Increase over Time.

The toxicity exerted by the antibiotic sulfadiazine on the growth of soil bacterial communities was studied in two agricultural soils for a period of 100 days. In the short-term (2 days of incubation), the effect of sulfadiazine on bacterial growth was low (no inhibition or inhibition <32% for a dose of 2000 mg·kg-1). However, sulfadiazine toxicity increased with time, achieving values of 40% inhibition, affecting bacterial growth in both soils after 100 days of incubation. These results, which were here observed for the first time for any antibiotic in soil samples, suggest that long-term experiments would be required for performing an adequate antibiotics risk assessment, as short-term experiments may underestimate toxicity effects.

Efficient sulfadiazine degradation via in-situ epitaxial grow of Graphitic Carbon Nitride (g-C3N4) on carbon dots heterostructures under visible light irradiation: Synthesis, mechanisms and toxicity evaluation.

The synthesis of environmental-friendly metal-free photocatalysts has great significance in photocatalytic technology. In this work, we firstly report the successful synthesis of in situ epitaxial growth of g-C3N4 on carbon dots through a facile thermal polymerization technique. Characterization and density functional theory (DFT) calculations were conducted to clarify the structure engineering and the electronic/chemical properties of the in-plane interconnected carbon dots/g-C3N4 (C-CN) heterostructures. With the optimal carbon dots content, the C-CN exhibited 3.2 times higher degradation rate for sulfadiazine (SDZ) than that of g-C3N4. Besides, the C-CN heterostructures displayed excellent stability and reusability in five consecutive cycles. The enhanced photocatalytic activity was related to the narrowed band gap and the local electronic density of valance band and conduction band orbitals of the unique plane heterostructures, corroborated by the spectroscopic characterizations and theoretical calculations. Photogenerated holes dominated the degradation of SDZ, while OH showed a negligible contribution. Moreover, DFT calculation succeeded to predict that the atoms with high Fukin index (f0) on SDZ molecule were more vulnerable to radicals attack. SDZ degradation pathway mainly included smiles-type rearrangement, SO2 extrusion, ring hydroxylation and SN bond cleavage processes. The eco-toxicity assessment revealed the generation of less toxic intermediates after photocatalysis. Our findings not only afford a new technique for constructing g-C3N4-based in-plane heterostructures with high and stable photocatalytic efficiency, but also highlight the feasible application of metal-free photocatalysts in environmental remediation.

A proteomic and biochemical investigation on the effects of sulfadiazine in Arabidopsis thaliana.

Animal manure or bio-solids used as fertilizers are the main routes of antibiotic exposure in the agricultural land, which can have immense detrimental effects on plants. Sulfadiazine (SDZ), belonging to the class of sulfonamides, is one of the most detected antibiotics in the agricultural soil. In this study, the effect of SDZ on the growth, changes in antioxidant metabolite content and enzyme activities related to oxidative stress were analysed. Moreover, the proteome alterations in Arabidopsis thaliana roots in response to SDZ was examined by means of a combined iTRAQ-LC-MS/MS quantitative proteomics approach. A dose-dependent decrease in leaf biomass and root length was evidenced in response to SDZ. Increased malondialdehyde content at higher concentration (2 µM) of SDZ indicated increased lipid peroxidation and suggest the induction of oxidative stress. Glutathione levels were significantly higher compared to control, whereas there was no increase in ascorbate content or the enzyme activities of glutathione metabolism, even at higher concentrations. In total, 48 differentially abundant proteins related to stress/stimuli response followed by transcription and translation, metabolism, transport and other functions were identified. Several proteins related to oxidative, dehydration, salinity and heavy metal stresses were represented. Upregulation of peroxidases was validated with total peroxidase activity. Pathway analysis provided an indication of increased phenylpropanoid biosynthesis. Probable molecular mechanisms altered in response to SDZ are highlighted.

Bioconcentration and ecotoxicity of sulfadiazine in the aquatic midge Chironomus riparius.

Although sulfadiazine (SDZ) is widespread in aquatic environments, information regarding the effects of SDZ on aquatic insects is still limited. In the present study, the bioconcentration and the effects of SDZ on the antioxidant system and the expression of endocrine and stress-related genes in Chironomus riparius larvae were investigated. The larvae were exposed to SDZ at the nominal concentrations of 2, 20 and 200 µg/L for 48 h. The results showed that SDZ was taken up by C. riparius despite presenting low bioconcentration factor values (0.99-3.92). In addition, superoxide dismutase activity was markedly reduced compared with the control group, whereas the levels of malondialdehyde were not significantly affected by SDZ. Moreover, the mRNA expression of genes related to heat shock proteins (Hsp70 and Hsp27) and ecdysone pathway (EcR and E74) were significantly up-regulated following all SDZ treatments. In aggregate, our work provides novel and interesting results regarding the potential biochemical and genetic effects of SDZ on freshwater insects.

Effects of biogas residues containing antibiotics on soil enzyme activity and lettuce growth.

To investigate the effects of biogas residues containing antibiotics on lettuce growth and soil enzyme activity, the antibiotics oxytetracycline and sulfadiazine were used as additives; two kinds of feedstocks including pig manure and straw were chosen. Results showed that the higher the concentrations of the antibiotics are, the stronger the inhibition of soil sucrase activity becomes. The activity of sucrose enzyme in the test group with a single concentration of 60 mg/kg was significantly lower than the group that was treated with 0 mg/kg. In the mass, the organic fertilizer containing antibiotics can improve the soil urease activity and increase the urease activity of the soil. In the growth phase, urease activity increased by 27% from 0.85 mg NH3-N/(g day) in the blank group to 1.08 NH3-N/(g day) in the 0-mg test group. During all growing stages of lettuce, though the effects of the biogas residues containing oxytetracycline and sulfadiazine on the soil catalase activity were not obvious, the field application could inhibit the activity of soil catalase to some extent.

Negative bottom-up effects of sulfadiazine, but not penicillin and tetracycline, in soil substitute on plants and higher trophic levels.

Veterinary antibiotics are widely used in livestock production and can be released to the environment via manure, affecting non-target organisms. Recent studies provide evidence that antibiotics can adversely affect both plants and insects but whether antibiotics in soil also affect trophic interactions is unknown. We tested whether antibiotics grown in sand as soil substitute with environmentally relevant concentrations of penicillin, sulfadiazine and tetracycline affect the survival of aphids feeding on plants (two crop and one non-crop plant species). Apera spica-venti, Brassica napus, and Triticum aestivum individuals were infested with aphids that were monitored over four weeks. We did not observe effects of penicillin or tetracycline on plants or aphids. However, sulfadiazine treatments reduced plant growth and increased mortality in the two tested grass species, but not in B. napus. Sulfadiazine subsequently decreased aphid density indirectly through reduced host plant biomass. We thus show that an antibiotic at realistic concentrations in a soil substitute can affect several trophic levels, i.e. plants and herbivores. This study contributes to the environmental risk assessment of veterinary antibiotics as it implies that their use potentially affects plant-insect interactions at environmentally relevant concentrations.

Co-exposure of Freshwater Microalgae to Tetrabromobisphenol A and Sulfadiazine: Oxidative Stress Biomarker Responses and Joint Toxicity Prediction.

Combined toxicity and oxidative stress biomarker responses were determined for tetrabromobisphenol A (TBBPA) and sulfadiazine (SDZ) to the unicellular green alga Scenedesmus obliquus. Concentration-response analyses were performed for single toxicants and for mixtures containing TBBPA and SDZ with two different mixture ratios. The effect concentrations and the observed effects of the mixtures were compared to the predictions of the joint toxicity by the concentration addition (CA) model and independent action (IA) model. Results showed that the observed joint toxicity was within the scope of the highest (TBBPA) and lowest (SDZ) toxicity observed for the individual components. Furthermore, co-exposure of S. obliquus to TBBPA and SDZ provided preliminary evidence that the mixtures induced oxidative stress leading to cell damage. The CA and IA models proved to be valid for the prediction of the joint toxicity of TBBPA and SDZ. This study highlights a combined environmental risk assessment for two emerging pollutants.

Bioaccumulation of sulfadiazine and subsequent enzymatic activities in Chinese mitten crab (Eriocheir sinensis).

The bioaccumulation of sulfadiazine and subsequent enzymatic activities in Chinese mitten crab (Eriocheir sinensis) were studied in microcosms, by exposing to 50, 100, 500 and 1000ng/L of sulfadiazine for 44days. An effective method for extracting sulfadiazine in crab tissues was established by modifying the cleanup method after ultrasound extraction, with improved recoveries of 61.8%, 93.7% and 100.5% in gill, muscle and liver samples. The results showed that sulfadiazine residues were all <3ng/g dry weight in different tissues, and that sulfadiazine bioaccumulation in crab was not dose-dependent. A significantly negative correlation was observed between acetylcholinesterase activity and the residue concentration of sulfadiazine during exposure to 50ng/L and 1000ng/L, and between alkaline phosphatase and sulfadiazine residues in the 100ng/L exposure group in the gill, suggesting that the two enzymes played an important role in the metabolism of sulfadiazine in crab.

Toxicity of sulfadiazine and copper and their interaction to wheat (Triticum aestivum L.) seedlings.

A pot experiment was carried out to investigate the single and combined effect of different concentrations of sulfadiazine (SDZ) (1 and 10mgkg-1) and copper (Cu) (20 and 200mgkg-1) stresses on growth, hydrogen peroxide (H2O2), malondialdehyde (MDA), antioxidant enzyme activities of wheat seedlings and their accumulation. High SDZ or Cu level significantly inhibited the growth of wheat seedlings, but the emergence rate was only inhibited by high SDZ level. The presence of Cu reduced the accumulation of SDZ, whereas the effect of SDZ on the accumulation of Cu depended on their concentrations. Low Cu level significantly increased the chlorophyll content, while high Cu level or both SDZ concentrations resulted in a significant decrease in the chlorophyll content as compared to the control. Additionally, H2O2 and MDA contents increased with the elevated SDZ or Cu level. The activities of superoxide dismutase, peroxidase and catalase were also stimulated by SDZ or Cu except for the aerial part treated by low Cu level and root treated by high SDZ level. The joint toxicity data showed that the toxicity of SDZ to wheat seedlings was generally alleviated by the presence of Cu, whereas the combined toxicity of SDZ and Cu was larger than equivalent Cu alone.

Reaction of antibiotic sulfadiazine with manganese dioxide in aqueous phase: Kinetics, pathways and toxicity assessment.

Sulfonamide antibiotics are often detected in terrestrial and aquatic environment, but little is known about abiotic degradation of these antibiotics. In the present study, the degradation of the sulfonamide antibiotic sulfadiazine by a synthesized δ-MnO2 was investigated. The initial reaction rate of sulfadiazine oxidized by manganese dioxide increased as the solution pH decreased by weakening electrostatic attraction between sulfadiazine and MnO2 and enhancing the reduction potential of MnO2. The presence of metal ions (Mn2+, Na+ and Ca2+), especially Mn2+, decreased the initial reaction rate by competitively adsorbing and reacting with MnO2. Two different products were identified during the reaction of sulfadiazine with MnO2 and the transformation of parent compound started with the formation of sulfadiazine radicals. Furthermore, toxicity assay results showed that the toxicity of products produced by bacteria decreased with elapse of reaction time. Results from the present study indicate that manganese dioxides in environmental matrix could be helpful in dissipation of sulfadiazine released into the environment.

Estimation of the toxicity of sulfadiazine to Daphnia magna using negligible depletion hollow-fiber liquid-phase microextraction independent of ambient pH.

The toxicity of ionizable organic compounds to organisms depends on the pH, which therefore affects risk assessments of these compounds. However, there is not a direct chemical method to predict the toxicity of ionizable organic compounds. To determine whether hollow-fiber liquid-phase microextraction (HF-LPME) is applicable for this purpose, a three-phase HF-LPME was used to measure sulfadiazine and estimate its toxicity to Daphnia magna in solutions of different pH. The result indicated that the sulfadiazine concentrations measured by HF-LPME decreased with increasing pH, which is consistent with the decreased toxicity. The concentration immobilize 50% of the daphnids (EC50) in 48 h calculated from nominal concentrations increased from 11.93 to 273.5 mg L-1 as the pH increased from 6.0 to 8.5, and the coefficient of variation (CV) of the EC50 values reached 104.6%. When calculated from the concentrations measured by HF-LPME (pH 12 acceptor phase), the EC50 ranged from 223.4 to 394.6 mg L-1, and the CV decreased to 27.60%, suggesting that the concentrations measured by HF-LPME can be used to estimate the toxicity of sulfadiazine irrespective of the solution pH.

Interspecific differences in growth response and tolerance to the antibiotic sulfadiazine in ten clonal wetland plants in South China.

Pollution caused by residual antibiotics is a worldwide environmental issue. Antibiotic residues often occur in aquatic ecosystems, posing threats to the health of aquatic organisms. The effects of antibiotic residues on the growth of crop plants and on human health are reasonably well known. However, less is known about antibiotic effects on wetland plants. Therefore, we studied the response and tolerance of ten clonal wetland plants grown in soil spiked with sulfadiazine at 10 mg kg(-1) (an environmentally relevant concentration) and 100 mg kg(-1). At 10 mg kg(-1), ramet number was the least affected trait, while root number was the most affected among plant species. Plant shoot and total biomass were reduced in all species except in Cyperus malaccensis var. brevifolius and Panicum repens. Chlorophyll content was reduced in Alocasia macrorrhiza, Saururus chinensis, and Commelina diffusa. In general, Panicum paludosum and C. malaccensis var. brevifolius showed the least reduction of growth parameters, whereas growth of both A. macrorrhiza and S. chinensis was severely reduced. At 100 mg kg(-1), negative responses occurred in all species. Comprehensive tolerance analysis revealed that P. paludosum and C. malaccensis var. brevifolius were the species most resistant to sulfadiazine. These species are potential candidates for sulfadiazine polluted wetland restoration. A. macrorrhiza and S. chinensis were the most susceptible species and they should be protected from sulfadiazine pollution. Relative plant shoot biomass and height were the most useful indicators for evaluating plant tolerance to sulfadiazine. Plant tolerance to sulfadiazine was associated with the differences of plants in height and shoot biomass.

Acute Toxicity and Environmental Risks of Five Veterinary Pharmaceuticals for Aquatic Macroinvertebrates.

Due to the high use of antibiotics and antiparasitics for the treatment of livestock, there is concern about the potential impacts of the release of these compounds into freshwater ecosystems. In this context, the present study quantified the acute toxicity of two antibiotics (sulfadiazine and sulfadimidine), and three antiparasitic agents (flubendazole, fenbendazole, ivermectin) for nine freshwater invertebrate species. These experiments revealed a low degree of toxicity for the sulfonamide antibiotics, with limited implications in the survival of all test species at the highest test concentrations (50 and 100 mg/L). In contrast, all three antiparasitic agents indicated on the basis of their acute toxicity risks for the aquatic environment. Moreover, chronic toxicity data from the literature for antiparasitics, including effects on reproduction in daphnids, support the concern about the integrity of aquatic ecosystems posed by releases of these compounds. Thus, these pharmaceuticals warrant further careful consideration by environmental risk managers.

Chemical oxidation of sulfadiazine by the Fenton process: kinetics, pathways, toxicity evaluation.

This paper investigated sulfadiazine oxidation by the Fenton process under various reaction conditions. The reaction conditions tested in the experiments included the initial pH value of reaction solutions, and the dosages of ferrous ions and hydrogen peroxide. Under the reaction conditions with pH 3, 0.25 mM of ferrous ion and 2 mM of hydrogen peroxide, a removal efficiency of nearly 100% was achieved for sulfadiazine. A series of intermediate products including 4-OH-sulfadiazine/or 5-OH-sulfadiazine, 2-aminopyrimidine, sulfanilamide, formic acid, and oxalic acid were identified. Based on these products, the possible oxidation pathway of sulfadiazine by Fenton's reagent was proposed. The toxicity evaluation of reaction solutions showed increased antimicrobial effects following the Fenton oxidation process. The results from this study suggest that the Fenton oxidation process could remove sulfadiazine, but also increase solution toxicity due to the presence of more toxic products.