Uptake of veterinary medicines from soils into plants

Medicines play an important role in the treatment and prevention of disease. Whereas the side effects on human and animal health resulting directly from treatment have been widely documented, only recently have the occurrence and fate of medicines in the environment and the potential consequences for human health been recognized as an issue warranting consideration. Medicines have been shown to be released to soils and to persist in the environment. This study was performed to investigate the potential for a range of veterinary medicines to be taken up from soil by plants used for human consumption and to assess the potential significance of this exposure route in terms of human health. Soil analyses indicated that, for selected substances, measurable residues of these are likely to occur in soils for at least 5 months following application of manure containing these compounds. Experimental studies on the uptake of veterinary medicines into carrot roots (tubers) and lettuce leaves showed that only florfenicol, levamisole, and trimethoprim were taken up by lettuces, whereas diazinon, enrofloxacin, florfenicol, and trimethoprim were detected in carrot roots. Measured concentrations in plant material were used to model potential adult human exposure to these compounds. Although exposure concentrations were appreciable in a few instances, accounting for approximately 10% of the acceptable daily intake values (ADI), all were lower than the ADI values, indicating that, at least for compounds with properties similar to those considered here, there is little evidence of an appreciable risk. This exposure route may, however, be important when veterinary medicines have a very low ADI, at which they elicit subtle effects over prolonged periods, or when exposure is occurring via a number of routes at once. Although degradation products (produced in the soil or the plant) were not measured, it is possible for some substances that these could increase the risks to consumers.     

The efficacy of three techniques to alleviate soil compaction at a restored sand and gravel quarry

Reinstated soil at restored sites often suffers from severe compaction which can significantly impede root development. Several methods, such as ripping and complete cultivation, are available to alleviate compaction that may occur as a result of soil reinstatement. This paper examines the effectiveness of the industry standard industrial ripper and a prototype modern ripper, the Mega‐Lift, in comparison with the recommended best practice method of complete cultivation. An investigation of the penetration resistance of the soil at a restored sand and gravel quarry was carried out using a cone penetrometer and a ‘lifting driving tool’ (dropping weight penetrometer) 3 years following cultivation. All the cultivation treatments reduced soil compaction to some degree compared with the untreated control. However, the penetration resistance values suggest that rooting would be restricted at relatively shallow depths in the plots cultivated using the industrial and Mega‐Lift ripper; penetration resistance exceeded 2 MPa within the first 0.33 m. Complete cultivation maintained penetration resistance values of less than 2 MPa within the depth limit of the penetrometer of 0.42 m. In addition, the results from the ‘lifting driving tool’ indicate that soils treated using complete cultivation remained significantly looser than those treated with the ripper to a depth of at least 0.80 m. The results demonstrate that complete cultivation remains the most effective method of alleviating soil compaction on restored sites, although it is recognized that its relatively high cost may restrict the uptake of the technique.     

Spatial modelling of the fraction of photosynthetically active radiation absorbed by a boreal mixedwood forest using a lidar–hyperspectral approach

The spatial variability of the fraction of photosynthetically active radiation absorbed by the canopy (fPAR) was characterized for a heterogeneous boreal mixedwood forest site located in northern Ontario, Canada, based on relationships found between fPAR and light detection and ranging (lidar) data over different canopy architectures. Estimates of fPAR were derived from radiation measurements made above the canopy at a flux tower and below-canopy radiation was measured across a range of species compositions and canopy architectures. Airborne lidar data were used to characterize spatial variability of canopy structure around the flux tower and a map of mean canopy chlorophyll concentration was derived from airborne hyperspectral imagery. Different volumes of lidar points for the locations directly above each photosynthetically active radiation (PAR) sensor were examined to determine if there is an optimal method of relating lidar returns to estimated fPAR values.The strongest correlations between mean lidar height and fPAR occurred when using points that fell within a theoretical cone which originated at the PAR sensor having a solid angle α = 55°. For diffuse conditions, the correlation (r) between mean lidar height versus fPAR × chlorophyll was stronger than between mean lidar height versus fPAR by 8% for mean daily fPAR and from 10 to 20% for diurnal fPAR, depending on solar zenith angle. For direct light conditions, the relationship was improved by 12% for mean daily fPAR and 12–41% for diurnal relationships.Linear regression models of mean daily fPAR × chlorophyll versus mean lidar height were used in conjunction with gridded lidar data and the canopy chlorophyll map to generate maps of mean daily fPAR for direct and diffuse sunlight conditions. Site average fPAR calculated from these maps was 0.79 for direct light conditions and 0.78 for diffuse conditions. When compared to point estimates of mean daily fPAR calculated on the tower, the average fPAR was significantly lower than the point estimate. Subtracting the direct sunlight fPAR map from the diffuse sunlight fPAR map revealed a distinct spatial pattern showing that areas with open canopies and relatively low chlorophyll (e.g., black spruce patches) have a higher fPAR under direct sunlight conditions, while closed canopies with higher chlorophyll (e.g., deciduous species) absorb more PAR under diffuse conditions. These findings have implications for scaling from point measurements at flux towers to larger resolution satellite imagery and addressing local scale heterogeneity in flux tower footprints.     

C and N natural abundance of the soil microbial biomass

Stable isotope analysis is a powerful tool in the study of soil organic matter formation. It is often observed that more decomposed soil organic matter is ¹³C, and especially ¹⁵N-enriched relative to fresh litter and recent organic matter. We investigated whether this shift in isotope composition relates to the isotope composition of the microbial biomass, an important source for soil organic matter. We developed a new approach to determine the natural abundance C and N isotope composition of the microbial biomass across a broad range of soil types, vegetation, and climates. We found consistently that the soil microbial biomass was ¹⁵N-enriched relative to the total (3.2 ‰) and extractable N pools (3.7 ‰), and ¹³C-enriched relative to the extractable C pool (2.5 ‰). The microbial biomass was also ¹³C-enriched relative to total C for soils that exhibited a C3-plant signature (1.6 ‰), but ¹³C-depleted for soils with a C4 signature (−1.1 ‰). The latter was probably associated with an increase of annual C3 forbs in C4 grasslands after an extreme drought. These findings are in agreement with the proposed contribution of microbial products to the stabilized soil organic matter and may help explain the shift in isotope composition during soil organic matter formation.     

The biomass of fungal mycelium in buried paleosols and surface soils of the steppe zone

The contents of fungal mycelium have been studied in paleosols of ancient archeological monuments and in surface soils within the steppe, dry steppe, and desert zones of European Russia, on the Stavropol, Privolzhskaya, and Ergeni uplands. The buried paleosols date back to the Bronze Age (4600–4500 and 4000–3900 BP), the Early Iron Age (1900–1800 BP), and the early 18th century (1719–1721). The fungal mycelium has been found in all these paleosols. The biomass of fungal mycelium varies from 2 to 124 μg/g of soil. The distribution patterns of fungal mycelium in the profiles of buried paleosols and surface soils have been identified. It is shown that the dark-colored mycelium is typical of the ancient paleosols. In some cases, the content of the dark-colored mycelium in them may reach 100% of the total mycelium biomass.     

Stability of copigmented anthocyanins and ascorbic acid in a grape juice model system

The stability of red grape anthocyanins (Vitis vinifera) was evaluated in a model juice system during normal (25 degrees C) and accelerated storage (35 degrees C) in the presence of ascorbic acid. Rosemary polyphenolic cofactors (0, 0.2, and 0.4% v/v) were evaluated as anthocyanin stabilizing agents. Cofactor addition resulted in concentration-dependent hyperchromic (up to 178%) and bathochromic (up to 23 nm) shifts, indicating a more intense red coloration of the models. Anthocyanin and ascorbic acid degradation followed first-order kinetics during storage. Results showed that copigmented treatments underwent a lower conversion of L-ascorbic acid into dehydroascorbic acid during storage when compared to the control, favorably impacting the vitamin retention of these models. Copigmentation did not affect anthocyanin degradation in the absence of ascorbic acid but in its presence aided to retain a higher anthocyanin content than the control. This study indicated that the addition of anthocyanin cofactors could be used to reduce the pigment and vitamin degradation while masking detrimental color changes in anthocyanin containing products.     

Dynamic headspace analysis of the release of volatile organic compounds from ethanolic systems by direct APCI-MS

Static equilibrium headspace was diluted with a stream of nitrogen to study the stability of the volatile headspace concentration. The headspace dilution profile of 18 volatile compounds above aqueous and ethanolic solutions was measured in real time using atmospheric pressure chemical ionization-mass spectrometry. Under dynamic conditions the volatiles headspace concentration above water solutions decreased readily upon dilution. The presence of ethanol helped to maintain the volatile headspace concentration when the ethanol solution concentration was above 50 mL/L. This effect was such that under dynamic conditions the absolute volatile concentration above an ethanolic solution was higher than that above an aqueous solution, contrary to results observed in equilibrium studies. The ratio of the headspace concentration of volatiles above ethanolic 120 mL/L and water solutions was correlated to their air/water partition coefficient.     

Residues of spiroxamine in grapes following field application and their fate from vine to wine

Dissipation of the fungicide spiroxamine in grapes of two vine varieties, Roditis and Cabernet Sauvignon, exposed to field treatments was evaluated. Vines of a grape vineyard located in central Greece were sprayed once or twice with a commercial formulation of the fungicide at 30 g a.i./hL. Residues in grapes, must, and wine were determined by gas chromatography/IT-MS after extraction with cyclohexane-dichloromethane (9:1), with a limit of quantitation 0.02 mg/kg in grapes and 0.012 mg/kg in wine. Under field conditions, spiroxamine dissipation on grapes was faster during the first 2 weeks and then slower to the sixth week. About 7 days after application, half of the initial spiroxamine concentration remained on the grapes; the respective proportion at 42 days was about 10%. At 14 and 35 days, residues were lower than 0.44 and 0.22 mg/kg, respectively, values below the maximum residue levels set by the European Union (1 mg/kg). Spiroxamine residues transferred from grapes into the must and through the vinification process into the wine were also studied. Mean transfer factors of 0.26 and 0.55 were found from grapes into wine for the wines obtained without maceration and with maceration, respectively. Residues in wine, prepared from grapes with a spiroxamine content of 0.11-0.20 mg/kg, varied from <0.026 to 0.09 mg/kg. Spiroxamine diastereomer B was found to dissipate slower than diastereomer A in the field as well as during the vinification process.     

Comparative effects of applying leguminous and non-leguminous green manures and inorganic N on biomass yield and nitrogen uptake in flooded rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

The beneficial role of green manures in rice production is generally ascribed to their potential of supplying plant nutrients, particularly nitrogen (N). However, the mechanisms through which green manures enhance the crop productivity are poorly understood. Pot experiments were conducted using a ¹⁵N-tracer technique: (1) to compare the biomass production potential of sesbania (Sesbania aculeata Pers.) and maize (Zea mays L.) as green manuring crops for lowland rice and (2) to compare the effect of the two types of green manure and inorganic N on the dry matter accumulation and N uptake by two rice (Oryza sativa L.) cultivars, viz. IR-6 and Bas-370. Although maize produced three times higher shoot biomass compared with sesbania, the latter showed higher N concentration; and thus the total N yield was similar in the two types of plants. Applying the shoot material of the two plants to flooded rice significantly enhanced the dry matter yield and N uptake by the two rice cultivars, the positive effects generally being more pronounced with sesbania than with maize amendment. The difference in the growth-promoting potential of the two plant residues was related more to an increased uptake of the native soil N rather than to their direct role as a source of plant-available N. A positive added nitrogen interaction (ANI) was observed due to both plant residues, the effect was much more pronounced with the application of sesbania than with maize residues. In both rice cultivars, inorganic N also caused a substantial ANI, particularly at higher application rate. Losses from the applied N were 2–3 times lower from sesbania, compared with maize treatment. Green manuring with sesbania also caused much lower N losses than the inorganic N applied at equivalent or higher rates. The overall benefit of green manuring to rice plants was higher than inorganic N applied at comparable rates. The two rice cultivars differed in their response to green manuring, IR-6 generally being more responsive than Bas-370.     

Cloning, functional expression, and characterization of cystatin in sesame seed

A cDNA fragment encoding cystatin, a cysteine protease inhibitor, was obtained from maturing sesame seeds. The clone was constructed in a nonfusion or fusion vector and then overexpressed in Escherichia coli. The recombinant cystatins were found in the soluble fraction of cell extract and were demonstrated to be functionally active in a reverse zymographic assay. The corresponding endogenous 22 kDa cystatin of low abundance in mature seeds was purified to homogeneity via a papain-coupling affinity column and confirmed by western blotting with antibodies against the recombinant cystatin. Both endogenous and recombinant cystatin proteins showed effective inhibitory activities against papain with K(i) values of 7.89 x 10(-8) M and 2.77 x 10(-8) M, respectively. Immunodetection indicated that cystatin was specifically expressed in maturing seeds and rapidly degraded in germination. Accordingly, zymographic and inhibition analyses showed that sesame cystatin could not inhibit the de novo synthesized proteases in germinating seeds. It is suggested that sesame cystatin may play a role in the regulation of endogenous cysteine proteases during seed maturation and germination.