Pharmacokinetics of the Recalcitrant Drug Lamotrigine: Identification and Distribution of Metabolites in Cucumber Plants.

Treated wastewater is an important source of water for irrigation. As a result, irrigated crops are chronically exposed to wastewater-derived pharmaceuticals, such as the anticonvulsant drug lamotrigine. Lamotrigine is known to be taken up by plants, but its plant-derived metabolites and their distribution in different plant organs are unknown. This study aimed to detect and identify metabolites of lamotrigine in cucumber plants grown for 35 days in a hydroponic solution by using LC-MS/MS (Orbitrap) analysis. Our data showed that 96% of the lamotrigine taken up was metabolized. Sixteen metabolites possessing a lamotrigine core structure were detected. Reference standards confirmed two; five were tentatively identified, and nine molecular formulas were assigned. The data suggest that lamotrigine is metabolized via N-carbamylation, N-glucosidation, N-alkylation, N-formylation, N-oxidation, and amidine hydrolysis. The metabolites LTG-N2-oxide, M284, M312, and M370 were most likely produced in the roots and were translocated to the leaves. Metabolites M272, M312, M314, M354, M368, M370, and M418 were dominant in leaves. Only a few metabolites were detected in the fruits. With an increasing exposure time, lamotrigine leaf concentrations decreased because of continuous metabolism. Our data showed that the metabolism of lamotrigine in a plant is fast and that a majority of metabolites are concentrated in the roots and leaves.

Application of biostimulant products and biological control agents in sustainable viticulture: A review.

Current and continuing climate change in the Anthropocene epoch requires sustainable agricultural practices. Additionally, due to changing consumer preferences, organic approaches to cultivation are gaining popularity. The global market for organic grapes, grape products, and wine is growing. Biostimulant and biocontrol products are often applied in organic vineyards and can reduce the synthetic fertilizer, pesticide, and fungicide requirements of a vineyard. Plant growth promotion following application is also observed under a variety of challenging conditions associated with global warming. This paper reviews different groups of biostimulants and their effects on viticulture, including microorganisms, protein hydrolysates, humic acids, pyrogenic materials, and seaweed extracts. Of special interest are biostimulants with utility in protecting plants against the effects of climate change, including drought and heat stress. While many beneficial effects have been reported following the application of these materials, most studies lack a mechanistic explanation, and important parameters are often undefined (e.g., soil characteristics and nutrient availability). We recommend an increased study of the underlying mechanisms of these products to enable the selection of proper biostimulants, application methods, and dosage in viticulture. A detailed understanding of processes dictating beneficial effects in vineyards following application may allow for biostimulants with increased efficacy, uptake, and sustainability.

Pharmacokinetics in Plants: Carbamazepine and Its Interactions with Lamotrigine.

Carbamazepine and lamotrigine prescribed antiepileptic drugs are highly persistent in the environment and were detected in crops irrigated with reclaimed wastewater. This study reports pharmacokinetics of the two drugs and their metabolites in cucumber plants under hydroponic culture, testing their uptake, translocation, and transformation over 96 h in single and bisolute systems at varying pH. Ruling out root adsorption and transformations in the nutrient solution, we demonstrate that carbamazepine root uptake is largely affected by the concentration gradient across the membrane. Unlike carbamazepine, lamotrigine is adsorbed to the root and undergoes ion trapping in root cells thus its translocation to the shoots is limited. On the basis of that, carbamazepine uptake was not affected by the presence of lamotrigine, while lamotrigine uptake was enhanced in the presence of carbamazepine. Transformation of carbamazepine in the roots was slightly reduced in the presence of lamotrigine. Carbamazepine metabolism was far more pronounced in the shoots than in the roots, indicating that most of the metabolism occurs in the leaves, probably due to higher concentration and longer residence time. This study indicates that the uptake of small nonionic pharmaceuticals is passive and governed by diffusion across the root membrane.

Composted biosolids and treated wastewater as sources of pharmaceuticals and personal care products for plant uptake: A case study with carbamazepine.

Irrigation with treated wastewater (TWW) and application of biosolids to arable land expose the agro-environment to pharmaceuticals and personal care products (PPCPs) which can be taken up by crops. In this project, we studied the effect of a carrier medium (e.g., biosolids and TWW) on plant (tomato, wheat and lettuce) uptake, translocation and metabolism of carbamazepine as a model for non-ionic PPCPs. Plant uptake and bioconcentration factors were significantly lower in soils amended with biosolids compared to soils irrigated with TWW. In soils amended with biosolids and irrigated with TWW, the bioavailability of carbamazepine for plant uptake was moderately decreased as compared to plants grown in soils irrigated with TWW alone. While TWW acts as a continuous source of PPCPs, biosolids act both as a source and a sink for these compounds. Moreover, it appears that decomposition of the biosolids in the soil after amendment enhances their adsorptive properties, which in turn reduces the bioavailability of PPCPs in the soil environment. In-plant metabolism of carbamazepine was found to be independent of environmental factors, such as soil type, carrier medium, and absolute amount implemented to the soil, but was controlled by the total amount taken up by the plant.

Characterization of breeding sites of Phlebotomus orientalis - the vector of visceral leishmaniasis in northwestern Ethiopia.

We studied breeding sites of Phlebotomus orientalis (Diptera: Psychodidae) the vector of visceral leishmaniasis in northern Ethiopia. Although numbers were rather small, 165 sand flies were captured emerging from vertisol cracks. The most productive breeding sites were cracked vertisols, dry river banks and close to trees. No sand flies were caught emerging from sandy clay loam soils in peri-domestic habitats but a few were captured emerging from gaps in a stone wall. Abiotic parameters in vertisols close to trees and in open field from which P. orientalis had emerged, were compared. Soil pH was slightly alkaline and salinity was low. Organic matter contents were similar in both sites. Temperatures and RH remained relatively stable near trees from the end of the rainy season through mid dry season, yet fluctuated markedly at the shallower depth in the open field. During the rainy season, cracks in the soil were sealed resulting in significant lowering of the oxygen concentrations near the tree. Gravimetric water content of soil near trees was lower than open field at shallow depth but similar deeper down. We conclude that ambient conditions suitable for sand fly larvae at shallow depths (45cm) are restricted to areas close to trees. However, deeper in vertisols (90cm) suitable conditions are apparently maintained throughout the dry season even in open fallow fields.

Irrigation of root vegetables with treated wastewater: evaluating uptake of pharmaceuticals and the associated human health risks.

To meet mounting water demands, treated wastewater has become an important source of irrigation. Thus, contamination of treated wastewater by pharmaceutical compounds (PCs) and the fate of these compounds in the agricultural environment are of increasing concern. This field study aimed to quantify PC uptake by treated wastewater-irrigated root crops (carrots and sweet potatoes) grown in lysimeters and to evaluate potential risks. In both crops, the nonionic PCs (carbamazepine, caffeine, and lamotrigine) were detected at significantly higher concentrations than ionic PCs (metoprolol, bezafibrate, clofibric acid, diclofenac, gemfibrozil, ibuprofen, ketoprofen, naproxen, sulfamethoxazole, and sildenafil). PCs in leaves were found at higher concentrations than in the roots. Carbamazepine metabolites were found mainly in the leaves, where the concentration of the metabolite 10,11-epoxycarbamazepine was significantly higher than the parent compound. The health risk associated with consumption of wastewater-irrigated root vegetables was estimated using the threshold of toxicological concern (TTC) approach. Our data show that the TTC value of lamotrigine can be reached for a child at a daily consumption of half a carrot (∼60 g). This study highlights that certain PCs accumulated in edible organs at concentrations above the TTC value should be categorized as contaminants of emerging concern.

Insights into the uptake processes of wastewater-borne pharmaceuticals by vegetables.

An increasing number of reports on plant uptake of pharmaceutical compounds (PCs) have been recently published, raising concerns of human exposure through dietary intake. In this study, PC uptake and translocation were evaluated in cucumber and tomato plants to elucidate the effects of PC physicochemical properties, soil type, and irrigation-water quality. Nonionic PCs were taken up and accumulated at higher levels in plants grown in soils of lower organic matter and clay content. While the concentration of most PCs in cucumber and tomato leaves were of similar order, their concentrations in the tomato fruit were much lower than in the cucumber fruit. This is related to differences in fruit physiology. Our data suggest that irrigation with treated wastewater reduces the bioavailability of acidic PCs for uptake by cucumber plants as compared to fresh water irrigation. This study sheds light on factors affecting the uptake of PCs by crops irrigated with treated wastewater, the governing role of PCs' physicochemical properties along with the physiological nature of the plant, soil properties and water quality that together determine uptake, translocation, and accumulation within plant organs. Occurrence of metabolites in plant suggests that PC metabolism has to be evaluated to reveal the total uptake.

Uptake of carbamazepine by cucumber plants--a case study related to irrigation with reclaimed wastewater.

Reclaimed wastewater is an important source of irrigation in semiarid and arid zones. Here we report data on carbamazepine (CBZ) uptake by cucumber plants in hydroponic culture and greenhouse experiments using different soil types irrigated with fresh water or reclaimed wastewater. Data obtained from the hydroponic culture experiments suggest that CBZ is mainly translocated by water mass flow, and thus it is concentrated and accumulated to the largest extent in the mature/older leaves. Carbamazepine concentration in cucumber fruits and leaves was negatively correlated with soil organic matter content. The concentrations of CBZ in the roots and stems were relatively low, and most CBZ in the plant (76-84% of total uptake) was detected in the leaves. A greenhouse experiment using fresh water and reclaimed wastewater spiked, or not, with CBZ at 1 µg L(-1) (typical concentration in effluents) revealed that CBZ can be taken up and bioaccumulated from its background concentration in reclaimed wastewater. Bioaccumulation factor (calculated as the ratio of CBZ concentration in the plant to that in the soil solution) for the fruits (0.8-1) was significantly lower than the value calculated for the leaves (17-20). This study emphasizes the potential uptake of active pharmaceutical compounds by crops in organic-matter-poor soils irrigated with reclaimed wastewater and highlights the potential risks associated with this agricultural practice.

Chemical fractionation of phosphorus in stabilized biosolids.

Three chemicals-ferrous sulfate (FeSul), calcium oxide (CaO), and aluminum sulfate (alum)-were applied at different rates to stabilize P in fresh, anaerobically digested biosolids (FBS) obtained from an activated sewage treatment plant. A modified Hedley fractionation procedure was used to assess P forms in these sludge-borne materials and in a biosolids compost (BSC) prepared from the same FBS. Each biosolids material exhibited a unique pattern of P distribution among fractions. The most available P forms, namely: (i) water-soluble P (WSP); (ii) membrane-P; and (iii) NaHCO(3)-P, were stabilized by small rates of each of the chemicals; but the P transformation into more stable forms depended on the type of chemical added. The stabilized P forms were enhanced by high rates of CaO and FeSul, but were reduced by high rates of alum. The organic P (P(o)) in the first three fractions of the FeSul- and alum-stabilized biosolids was enhanced by the chemical addition, and P(o) transformation from NaOH-P(o) into NaHCO(3)-P(o) was found in calcium-stabilized biosolids. A positive relationship was found between NaHCO(3)-P(o) and the NaHCO(3)-extracted organic C in all chemically stabilized biosolids. One-step extraction by NaHCO(3) or NaOH underestimated P extraction compared to the stepwise extraction. The reported results are consistent with solid-state P speciation reported earlier and contribute important information for optimizing biosolids stabilization to reduce P loss after incorporation in soils and for maximizing soil capacity to safely store pre-stabilized biosolids.

Solid phosphorus phase in aluminum- and iron-treated biosolids.

Stabilization of phosphorus (P) in sewage sludge (biosolids) to reduce water-soluble P concentrations is essential for minimizing P loss from amended soils and maximizing the capacity of the soil to safely serve as an outlet for this waste material. The chemical form at which P is retained in biosolids stabilized by Al(2)(SO(4))(3) x 18H(2)O (alum) or FeSO(4) x 7H(2)O (FeSul) was investigated by scanning electron microscopy (SEM) equipped with energy-dispersive X-ray elemental spectrometry (EDXS) and by X-ray diffraction (XRD). Both treatments resulted in the formation of a Ca-P phase, probably brushite. Phosphorus was further retained in the alum-treated biosolids by precipitation of an Al-P phase with an Al/P molar ratio of about 1:1, while in the FeSul-treated biosolids, P was retained by both precipitation with Fe/P molar ratios of 1:1 or 1.5:1, and by adsorption onto newly formed Fe hydroxides exhibiting an Fe/P molar ratio of up to 11:1. All of these mechanisms efficiently reduced P solubility and are crucial in biosolids environmentally safe agronomic beneficial use for this waste product; however, each P phase formed may react differently in the amended soil, depending on soil properties. Thus, the proper P stabilization method would depend on the target soil.

Water-soluble and solid-state speciation of phosphorus in stabilized sewage sludge.

Three chemicals, ferrous sulfate (Fe-sul), calcium oxide (CaO), and aluminum sulfate (alum), were used to stabilize phosphorus (P) in fresh, anaerobically digested sewage sludge (FSS). The chemically stabilized sludge materials and biosolids compost (BSC) were compared with the FSS with respect to water-soluble phosphorus (WSP) content in its inorganic (WSP(i)) and organic (WSP(o)) forms as well as water-soluble organic carbon (DOC). Solid-state P speciation was further probed by X-ray diffraction (XRD) and scanning electron microscopy (SEM) equipped with energy-dispersive X-ray elemental spectrometry (EDXS). Water-soluble P was effectively controlled by a wide range of Fe-sul or CaO additions to the sludge (Ca to P ratio = 3.47-17.72, Fe to P ratio = 1.01-16.53), but by only a narrow range (Al to P ratio = 1.04-2.87) of alum addition. The WSP content in the BSC was also depressed, but to a lesser extent. The pH in the treated sludge ranged from 3.0 to 12.5 and served as a key factor to control P chemistry. No correlation was observed between DOC and WSP(o). No crystallized Ca-P minerals were detected in the CaO-stabilized sludge, but brushite crystallization seemed to be obtained by low addition of Fe-sul and alum. Variscite and strengite crystallization was obtained following high addition of Fe-sul or alum, as detected by XRD and SEM-EDXS. Adsorption of P by newly formed Fe-hydroxide seems to play an important role in the Fe-sul-stabilized sludge. We concluded that administration of the tested chemicals at the proper rate can effectively reduce the hazard of P release and leaching from sludge.

Manganese nutrition effects on tomato growth, chlorophyll concentration, and superoxide dismutase activity.

The effects of Mn nutrition of tomato (Lycopersicon esculentum Mill.) seedlings on Mn-, Fe- and CuZn-superoxide dismutase (SOD, EC 1.15.1.1) enzymatic activities, metal translocation, chlorophyll concentration, and plant growth were tested using a bioassay system consisting of chelator-buffered nutrient culture with Mn2+ activities set to pMn (-log activity of Mn2+) of 6.6, 7.6, 8.6, and 9.6. The two middle levels resulted in optimal plant growth, whereas the two extreme levels resulted in a gradual decrease in chlorophyll concentration and slower plant growth. At the end of the experiment, 26 days after transfer to the Mn treatments, significant differences in shoot Mn concentration were manifested, from 10.5 mg kg(-1) in plants grown in pMn 9.6 to 207.4 mg kg(-1) in plants grown in pMn 6.6. Other element concentrations in the leaf suggest that growth inhibition and chlorophyll synthesis were affected primarily by manganese deficiency and excess. Twenty days after transfer of plants to the Mn treatments Mn-, Fe- and CuZn-SOD activities were assayed in young expanded leaf tissues by electrophoresis running gel. Whereas chloroplastic CuZn-SOD activity did not differ among Mn treatments, the cytosolic CuZn-SOD and mitochondrial Mn-SOD activities increased in both Mn-excess and Mn-deficient plants.