Suchart Upatham

Phytoextraction and Accumulation of Lead from Contaminated Soil by Vetiver Grass: Laboratory and Simulated Field Study

A soil-culture study was conducted to investigate the phytoextraction of lead (Pb) in two species of vetiver grass (Vetiveria zizanioides and V. nemoralis) irrigated with an increasing level of Pb(NO3)2 (5, 7, 9 and 11 g L-1) for 12 weeks. In a laboratory study, the removalof lead from soil was correlated with lead accumulation by roots and shoots of both species of vetiver grass. High concentration of lead (9—11 g L-1) resulted in decrease of growth, total chlorophyllcontent and biomass of V. zizanioides, while V. nemoralis died after one week of application. Toxicity symptoms (e.g., burning leaf margins, shoot die back) occurred in vetiver grass at a high concentration of lead. Based on the data V. zizanioides tolerated and accumulated the greatest amount of lead most effectively. A simulated field experiment was conducted to examine the efficiency of vetiver grass in removing lead from contaminated soil. The vetiver grasses, V. zizanioides and V. nemoralis, were grown in soil contaminated with Pb(NO3)2 (5, 7, 9, and 11 g L-1) for 3 months. The removal of lead from soil was correlated with lead accumulation by roots and shoots of both grass species. The grass roots took up more lead than the shoots. V. zizanioides could uptake more lead from soil than V. nemoralis. The effects of lead on the biomass of V. zizanioides and V. nemoralisshowed that in both species, the biomass was decreased when the lead concentration was increased. In comparison, V. zizanioidesshowed greater biomass than V. nemoralis.

Effects of Soil Amendments and EDTA on Lead Uptake by Chromolaena Odorata: Greenhouse and Field Trial Experiments

Greenhouse and field trial experiments were performed to evaluate the use of Chromolaena odorata with various soil amendments for phytoextraction of Pb contaminated soil. Pb mine soils contain low amount of nutrients, so the additions of organic (cow manure) and inorganic (Osmocote and NH4NO3 and KCl) fertilizers with EDTA were used to enhance plant growth and Pb accumulation. Greenhouse study showed that cow manure decreased available Pb concentrations and resulted in the highest Pb concentration in roots (4660 mg kg−1) and shoots (389.2 mg kg−1). EDTA increased Pb accumulation in shoots (17-fold) and roots (11-fold) in plants grown in soil with Osmocote with Pb uptake up to 203.5 mg plant−1. Appli- cation of all fertilizers had no significant effects on relative growth rates of C. odorata. Field trial study showed that C. odorata grown in soil with 99545 mg kg−1 total Pb accumulated up to 3730.2 and 6698.2 mg kg−1 in shoots and roots, respectively, with the highest phytoextraction coefficient (1.25) and translocation factor (1.18). These results indicated that C. odorata could be used for phytoextraction of Pb contaminated soil. In addition, more effective Pb accumulation could be enhanced by Osmocote fertilizer. However, the use of EDTA in the field should be concerned with their leaching problems.

Effect of Corn Plant on Survival and Phenanthrene Degradation Capacity of Pseudomonas Sp. UG14Lr in Two Soils

A study was undertaken to assess if corn (Zea mays L.) can enhance phenanthrene degradation in two soils inoculated with Pseudomonas sp. UG14Lr. Corn increased the number of UG14Lr cells in both soils, especially in the acidic soil. Phenanthrene was degraded to a greater extent in UG14Lr-inoculated or corn-planted soils than uninoculated and unplanted soils. The spiked phenanthrene was completely removed within 70 days in all the treatments in slightly alkaline soil. However, in acidic soil, complete phenanthrene removal was found only in the corn-planted treatments. The shoot and root lengths of corn grown in UG14Lr-inoculated soils were not different from those in non-inoculated soil between the treatments. The results showed that in unplanted soil, low pH adversely affected the survival and phenanthrene degradation ability of UG14Lr. Planting of corn significantly enhanced the survival of UG14Lr cells in both the bulk and rhizospheric soil, and this in turn significantly improved phenanthrene degradation in acidic soil. Re-inoculation of UG14Lr in the acidic soil increased the number of UG14Lr cells and enhanced phenanthrene degradation in unplanted soil. However, in corn-planted acidic soils, re-inoculation of UG14Lr did not further enhance the already active phenanthrene degradation occurring in both the bulk or rhizospheric soils.

Toxic effects of selenium inhalation: acute damage of the respiratory system of mice

Accidental inhalation of selenium by humans has been associated with damage of respiratory tissues that is lacking a detailed histological definition. We have investigated the natural history of injury to the tracheal epithelium and lungs induced by a single intratracheal instillation of CD-1 mice with two different doses of dimethyl selenide (0.05 and 0.1 mg Se/kg of body weight). The animals were sacrificed 1, 7, 14, and 28 days after the single selenium treatment. Samples of the trachea and lungs were studied by light microscopy. The tracheal epithelium showed loss of cilia and acute necrosis that was followed by metaplastic transformation. Edema and diffuse alveolar damage was observed in the lungs. Our data suggest that: i) severity of respiratory lesions caused by selenium is dose dependent; ii) selenium causes transient metaplastic transformation of the tracheal epithelium; iii) chronic inflammation and increased thickness of alveolar septa occur in the lungs; iv) 4 weeks after selenium treatment, mice recover from the tracheal injury, whereas no amelioration of pulmonary lesions was observed.

In vivo ingestion of heavy metal particles of Se, Hg and W by murine macrophages. A study using scanning electron microscopy coupled with X-ray microanalysis.

Several heavy metals that are currently employed in industry may become polluters of work and natural environments. As particulate matter, heavy metals are suitable for entering the human body through the respiratory and digestive systems. They often end up inside phagocytes; the size of the microscopic particles modulates both their phagocytosis, and the physiology of macrophages. Here we have adopted an experimental model to investigate the ingestion of particles of three industrial heavy metals (Se, Hg, W) by murine peritoneal macrophages in vivo. The phagocytes were studied by scanning electron microscopy coupled with X-ray elemental microanalysis (SEM-XRM), a method that allows specific identification of Se, W and Hg in cells at high resolution. We found that Hg that was taken up by macrophages was organized into small, round particles (0.319 / 0.14 mm). This was in contrast with the larger size of intracellular particles of Se (2.379 / 1.84 mm) or W (1.759-1.34 mm). Ingested particles of Se and W, but not Hg, often caused bulging of the cell surface of macrophages. We conclude that particulate matters of Se, W and Hg are organized in particles of different size inside macrophages. This size difference is likely to be associated with distinct phlogistic activities of these heavy metals, Se and W causing a milder inflammatory reaction than Hg.

Tubular Kidney Damage and Centrilobular Liver Injury after Intratracheal Instillation of Dimethyl Selenide

Accidental inhalation of selenium (Se) derivatives, such as dimethyl selenide (DMSe), has been associated with damage of respiratory tissues. However, systemic effects of inhaled Se have not been thoroughly established. We have investigated whether mouse kidney and liver show cellular pathology as a result of a single intratracheal instillation of two different doses of DMSe (0.05 and 0.1 mg Se/kg BW). The animals were sacrificed 1, 7, 14, and 28 days after either 1 of the 2 DMSe treatments; samples were studied by light microscopy. Instillation of the low DMSe dose resulted in acute and transient tubular disease of the kidney expressed by swelling and vacuolation of epithelial cells of proximal tubules; in some mice, tubular necrosis was observed. After 14 days of the DMSe treatment, these lesions were ameliorated and, by day 28, the kidney tubular epithelium depicted a normal morphology. The same low dose of DMSe caused sustained damage to centrilobular hepatocytes characterized by swollen and vacuolized liver cells. After the instillation of the high DMSe dose, the mice presented sustained liver and kidney focal necrosis. Our data suggest that inhalation of DMSe results in: (i) acute tubular injury of the kidney and damage to centrilobular liver cells and (ii) this systemic pathology induced by DMSe is a dose-dependent phenomenon.

Acute Pulmonary Inflammation Induced by Lung Overloading with Selenium Particles: Leukocyte Response and In Situ Detection of Selenium at High Resolution

The kinetics of the acute inflammatory response of the lung was triggered in CD-1 mice by a single intratracheal instillation of a large amount of Se (10 mg); it was studied by quantitative cytology of bronchoalveolar lavage samples, light microscopy, and scanning electron microscopy coupled with x-ray elemental microanalysis. Bronchoalveolar lavage leukocytes were mostly neutrophils and increased from 12 to 24 h of Se treatment and decreased at 72 h. Only less than half of the granulocytes showed ingested Se particles; in contrast, virtually all BAL macrophages contained Se particles. Scanning electron microscopy coupled with X-ray elemental microanalysis revealed that the intracellular Se particles were heterogeneous in size (diameters from 0.4 and up to 14 μm), and that Se inclusions were sometimes accumulated at a pole of the cell. At 72 h after instillation of the particles, Se-loaded alveolar macrophages were migrated in the interstitial space of the alveoli. Se-positive regions had a focal distribution in the lung; accumulation of inflammatory cells erased the alveolar architecture of these areas of the deep lung. Our data indicates that Se overloading of the lung results in: (1) an acute inflammatory response that is dominated by neutrophils; (2) early removal of Se done mostly by alveolar macrophages, and (3) formation of focal areas of invasion of the lung parenchyma by inflammatory infiltrates.

Changes in Bronchoalveolar Lavage Cells after Intratracheal Instillation of Dimethyl Selenide in Mice

CD-1 mice were exposed to a single intratracheal instillation of either 0.025 or 0.075 mg Se/kg wt of dimethyl selenide (DMSe). They were studied over 4 weeks to define the cellular inflammatory response of the airways to DMSe. Bronchoalveolar (BAL) lavage was used to collect the DMSe-induced inflammatory exudates. The DMSe instillation resulted in phlogistic responses that had the neutrophil as the main leukocyte; they were present in BAL samples, mostly at days 1 and 7. Macrophages were also increased during DMSe-induced inflammation. The lower DMSe dose resulted in an inflammatory reaction lasting for 2 weeks. Mice treated with the higher DMSe dose still showed elevated numbers of neutrophils and macrophages 4 weeks after instillation. DMSe did not change the number of lymphocytes harvested from the airways. An early increase in total protein of BAL, and late enhancement in lactate dehydrogenase was observed in mice treated with the high DMSe dose. We conclude that inhalation of DMSe triggers a moderate and dose-dependent inflammatory reaction in the mouse airways, and that this phlogistic reaction is likely to participate in the damage of respiratory epithelia that occurs upon DMSe inhalation.