BACKGROUND: Stevensite and biochar were investigated to covalently immobilize laccase from Myceliophthora thermophila (MtL) and Pleurotus eryngii (PeL) through the sequential application of aminopropyltriethoxysilane and glutaraldehyde. The immobilized preparations were tested to remove three tetracyclines and six sulfonamides at 0.1 mmol L −1 of each antibiotic. Degradation experiments were conducted both in the absence and in the presence (0.2 mmol L −1 ) of ABTS, 1-hydroxybenzotriazol (HBT), syringaldehyde or violuric acid. The residual antibiotic activity was tested towards five bacterial species and a bacterial consortium from wastewater. RESULTS: Higher values of activity yields (74% and 70.3%) and catalytic capabilities (1426 and 1405 IU g −1 ) were obtained with PeL on stevensite and biochar than with MtL. Stevensite enabled higher reusability and storage stability than biochar. Best removals of tetracyclines and sulfonamides were obtained with immobilized-laccase systems coupled to ABTS or syringaldehyde. Immobilized-laccase/ABTS systems removed 100% of tetracyclines while only chlortetracycline was completely removed in the presence of syringaldehyde. With ABTS, the most effectively removed sulfonamides were sulfathiazole and sulfadiazine (up to 100% and 54%), while syringaldehyde best supported the removal of sulfanilamide, sulfamethazine and sulfamethoxazole (up to 42%, 45% and 46%, respectively). In some cases, an effective antibiotics removal led to either low or no residual antibiotic activity. CONCLUSION: MtL and PeL were immobilized successfully on biochar and stevensite. The addition of either ABTS or syringaldehyde enhanced significant removals, up to 100%, of tetracyclines and sulfonamides by the immobilized laccase systems. Noteworthy, biochar-immobilized laccases/ABTS led to complete suppression of the antibiotic activity of tetracyclines.
The presence of antibiotics in crops is mainly caused by their irrigation with reclaimed wastewater and by the use of organic amendments of animal origin. During this work, the fate of sulfonamide antibiotics in tomato crop has been assessed in two commercial greenhouses located in Almería (Spain) irrigated with reclaimed wastewater. Samplings were made annually for two years. Sulfonamides in several parts of the plant (roots, leaves and fruits) as well as reclaimed wastewater, amendments and soils were analyzed by UHPLC-MS/MS. The results showed that sulfonamides accumulated in soils (sulfamethoxazole between 2 and 14 µg Kg−1; sulfadiazine, sulfathiazole, sulfapyridine, sulfamerazine and sulfadimethoxine in concentrations below 1 µg Kg−1) were in the reclaimed wastewater at concentrations in the ng L−1 range. Their distribution in plants depended on the sulfonamide. The sulfonamides detected in tomato were sulfadiazine, sulfapyridine, sulfamethazole, sulfamethoxazole and sulfadimethoxine. Sulfamethoxazole was the antibiotic with highest concentration in tomato fruit, exceeding 30 µg Kg−1. All sulfonamides were below the Acceptable Daily Intake, however, further studies and legislation are needed to assure food safety.
A double strategy based on the removal of sulfonamide antibiotics by Pleurotus ostreatus and adsorption on spent mushroom substrate was assessed to reclaim contaminated wastewater. P. ostreatus was firstly tested a in liquid medium fortified with five sulfonamides: sulfamethoxazole, sulfadiazine, sulfathiazole, sulfapyridine and sulfamethazine, to evaluate its capacity to remove them, to test for any adverse effects on fungal growth and for any reduction in residual antibiotic activity. P. ostreatus was effective in removing sulfonamides up to 83 to 91 % of the applied doses over 14 days. The antibiotic activity of the sulfonamide residues was reduced by 50 %. Sulfamethoxazole transformation products by laccase were identified and the degradation pathway was proposed. In addition, P. ostreatus growth on a semi-solid medium of spent mushroom substrate and malt extract agar was used to develop a biofilter for the removal of sulfonamides from real wastewater. The biofilter was able to remove more than 90% of the sulfonamide concentrations over 24 hours by combining adsorption and biodegradation mechanisms.
The aim of this study was to determine the effects of new Micro Carbon Technology (MCT®) fertilizers based on humic acids biologically digested on the growth and development of pepper plants. In this work, the biostimulant effect of MCT® fertilizers was compared to conventional mineral fertilizers. In order to evaluate MCT® fertilizers, a previous chemical characterization (infrared spectroscopy, liquid chromatography and mass spectrometry) of seven MCT® fertilizers was performed. Two fertilization tests of pepper plants were carried out in hydroponic conditions, where the fertilization and the age of the plants were studied in order to evaluate the specific effects on roots and leaves. Plant weight and foliar analysis (chlorophyll indices and nutrients) have been determined. Transmission electron microscopy was used to visualize the morphological differences in the root and leaf cells. Comparison between conventional and MCT® based fertilizers showed that, with the MCT® fertilizers, the plant is exposed to the presence of free amino acids (Glycine and Alanine), polyphenols and humic substances. Although no significant differences were found in plant mass production, the plants fertilized with MCT® products presented better nutritional status than plants treated with conventional fertilization in terms of nutrient content in leaves. Important morphological differences in root cells were found. A large central vacuole that represented the 68–83% of the total root cell area was shown if the MCT® products were used, suggesting significant changes of membrane permeability in terms of water adsorption and consequently nutrient storage. The morphological differences observed in the root cells were more noticeable in adult plants.
Trichloroethylene (TCE) is a proven carcinogenic chlorinated organic compound widely used as a solvent in industrial cleaning solutions; it is easily found in the soil, air, and water and is a hazardous environmental pollutant. Most studies have attempted to remove TCE from air and water using different anaerobic bacteria species. In addition, a few have used white-rot fungi, although there are hardly any in soil. The objective of the present work is to assess TCE removal efficiency using two species of the genus Pleurotus that have not been tested before: Pleurotus ostreatus and Pleurotus eryngii, growing on a sandy loam soil. These fungi presented different intra- and extracellular enzymatic systems (chytochrome P450 (CYP450), laccase, Mn peroxidase (MnP)) capable of aerobically degrading TCE to less harmful compounds. The potential toxicity of TCE to P. ostreatus and P. eryngii was firstly tested in a TCE-spiked liquid broth (70 mg L−1 and 140 mg L−1) for 14 days. Then, both fungi were assessed for their ability to degrade the pollutant in sandy loam soil spiked with 140 mg kg−1 of TCE. P. ostreatus and P. eryngii improved the natural dissipation of TCE from soil by 44%. Extracellular enzymes were poorly expressed, but mainly in the presence of the contaminant, in accordance with the hypothesis of the involvement of CYP450.
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