Phytoremediation technology for recovery of Ni by Acacia plants in association with Bacillus amyloliquefaciens isolated from E-waste contaminated site.

Electronic waste (e-waste) illegally disposal in Thailand is becoming more widespread. A sustainable metal recovery technology is needed. A phytotechnology called "phytomining" of metals such as nickel (Ni) is a promising technology providing a sustainable solution to the growing e-waste problems. This study investigated the ability of Acacia species in association with e-waste site isolated, plant growth-promoting rhizobacteria (PGPR), Bacillus amyloliquefaciens. Acacia mangium accumulated higher Ni in their tissues when Ni concentrations in soil were lower than 200 mg kg-1. The inoculation of PGPR B. amyloliquefaciens enhanced Ni uptake and accumulation in the leaves, stem, and root. The results showed that the highest Ni concentration was found in the root ash (825.50 mg kg-1) when inoculated plants were grown in soil containing 600 mg kg-1 Ni. Hence, the Ni recovery process and mass balance were performed on root ashes. The highest Ni recovery was 91.3% from the acid (H2SO4) leachate of the ash of inoculated plant treated with 600 mg kg-1 Ni. This demonstrates the feasibility of PGPR-assisted phytomining from Ni-contaminated soil. Phytomining of Ni from any e-waste contaminated sites using Acacia mangium in combination with B. amyloliquefaciens can promote plant growth and improve the uptake of Ni.

Phytomining from electronic waste is an appealing technology that can provide a long-term waste management strategy while valuable trace metals can be recovered. In this study, we evaluated the nickel phytomining ability of Acacia mangium in association with PGPR Bacillus amyloliquefaciens. The results from this study showed that Ni recovery from phytomass using A. mangium with B. amyloliquefaciens can be further improved leading to a sustainable waste management strategy.

A Place-Based Conceptual Model (PBCM) of Neotricula aperta/Schistosoma mekongi habitat before and after dam construction in the Lower Mekong River.

In 1971, scientists from Mahidol University in Thailand and the Smithsonian Institution in the USA formed a research team to study a new species of Schistosoma in the Mekong River in Thailand and Laos. The studies, completed during 1971-1973, prior to the construction of any dams or restrictions to the natural flow regime of the Mekong River, provide a unique description of the natural ecological state of the river that can serve as a baseline for current research. The natural transmission of Schistosoma japonicum, Mekong Strain, was first reported on Khong Island, Laos in 1973 using sentinel mice. The first detailed description of the habitat ecology of the snail vector Neotricula aperta was done on-site in 1971 simultaneously with that research and is unique in providing the only description of the river shoreline habitat before any dams were built and any alteration of the natural flow regime was in place. Aggregating current information in a Place-Based Conceptual Model (PBCM) as an organizing template, along with current habitat models that combine ecological data with e-flows, can be developed and used as a tool to predict suitable habitats for snails. The natural flow regime of the Mekong River prior to any impoundments is described with current updates on the potential impacts of climate change and dams with flow-related snail habitat characteristics, including sediment drift and water quality. The application of the PBCM to describe and compare descriptive information on current and potential future N. aperta/S. mekongi habitat is discussed.

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.

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). Application 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.

Plant-enhanced phenanthrene and pyrene biodegradation in acidic soil.

A study was undertaken to assess if corn plant (Zea may L.) maybe able to enhance the degradation of phenanthrene and pyrene in acidic soil inoculated with a bacterial strain (Pseudomonas putida MUB1) capable of degrading polycyclic aromatic hydrocarbons (PAHs). Planting with corn, inoculating with MUB1, ora combination of the two were found to promote the degradation of phenanthrene and pyrene in acidic soil at different rates. In the presence of corn plants, the rates of phenanthrene and pyrene removal were 41.7 and 38.8% in the first 10 days, while the rates were 58.8 and 53.6%, respectively in the treatment which received MUB1 only. After 60 days, the corn + MUB1 treatment led to the greatest reduction in both phenanthrene and pyrene biodegradation (89 and 88.2%, respectively). In control autoclaved soil, the rates of phenanthrene and pyrene removal were 14.2 and 28.7%, respectively while in non-autoclaved soil, the rates were 68.7 and 53.2%, respectively. These results show that corn, which was previously shown to grow well in PAH-contaminated acidic soil, also can enhance PAH degradation in such soil. Inoculation with a known PAH degrader further enhanced PAH degradation in the presence of corn.

Quantitative detection of the oil-degrading bacterium Acinetobacter sp. strain MUB1 by hybridization probe based real-time PCR.

Quantitative detection of the oil-degrading bacterium Acinetobacter sp. strain MUB1 was performed using the SoilMaster() DNA Extraction Kit (Epicentre, Madison, Wisconsin) and hybridization probe based real-time PCR. The detection target was the alkane hydroxylase gene (alkM). Standard curve construction showed a linear relation between log values of cell concentrations and real-time PCR threshold cycles over five orders of magnitude between 5.4+/-3.0x10(6) and 5.4+/-3.0x10(2)CFUml(-1) cell suspension. The detection limit was about 540CFUml(-1), which was ten times more sensitive than conventional PCR. The quantification of Acinetobacter sp. strain MUB1 cells in soil samples resulted in 46.67%, 82.41%, and 87.59% DNA recovery with a detection limit of 5.4+/-3.0x10(4)CFUg(-1) dry soil. In this study, a method was developed for the specific, sensitive, and rapid quantification of the Acinetobacter sp. strain MUB1 in soil samples.

Enhanced biodegradation of anthracene in acidic soil by inoculated Burkholderia sp. VUN10013.

The ability of Burkholderia sp. VUN10013 to degrade anthracene in microcosms of two acidic Thai soils was studied. The addition of Burkholderia sp. VUN10013 (initial concentration of 10(5) cells g(-1) dry soil) to autoclaved soil collected from the Plew District, Chanthaburi Province, Thailand, supplemented with anthracene (50 mg kg(-1) dry soil) resulted in complete degradation of the added anthracene within 20 days. In contrast, under the same test conditions but using autoclaved soil collected from the Kitchagude District, Chanthaburi Province, Thailand, only approximately 46.3% of the added anthracene was degraded after 60 days of incubation. In nonautoclaved soils, without adding the VUN10013 inocula, 22.8 and 19.1% of the anthracene in Plew and Kitchagude soils, respectively, were degraded by indigenous bacteria after 60 days. In nonautoclaved soil inoculated with Burkholderia sp. VUN10013, the rate and extent of anthracene degradation were considerably better than those seen in autoclaved soils or in uninoculated nonautoclaved soils in that only 8.2 and 9.1% of anthracene remained in nonautoclaved Plew and Kitchagude soils, respectively, after 10 days of incubation. The results showed that the indigenous microorganisms in the pristine acidic soils have limited ability to degrade anthracene. Inoculation with the anthracene-degrading Burkholderia sp. VUN10013 significantly enhanced anthracene degradation in such acidic soils. The indigenous microorganisms greatly assisted the VUN10013 inoculum in anthracene degradation, especially in the more acidic Kitchagude soil.

Phytotoxicity assay of crop plants to phenanthrene and pyrene contaminants in acidic soil.

Four selected plants (corn, groundnut, cow pea, and mungbean) were tested for their ability to germinate and grow in an acidic soil contaminated with phenanthrene or pyrene, two typical polycyclic aromatic hydrocarbons (PAHs). The growth of corn root was the least sensitive to, but its germination rate was the lowest in the presence of, contaminants. Among the legumes, the growth of groundnut root was better than others. Corn and groundnut were selected to further test their ability to tolerate a mixture of phenanthrene and pyrene in the acidic soil. The presence of both PAHs led to a greater decrease in the lengths of shoot and root of groundnut than phenanthrene or pyrene alone, but the lengths of shoot and root of corn were decreased to a similar extent as when phenanthrene or pyrene was present alone. The growth of corn root was also better than that of groundnut root when they were grown in oil-contaminated soil. Based on these results, we conclude that corn is the most suitable to be grown in PAH-contaminated acidic soil.

Batch and continuous packed column studies of cadmium biosorption by Hydrilla verticillata biomass.

The removal of heavy metal ions by the nonliving biomass of aquatic macrophytes was studied. We investigated Cd biosorption by dry Hydrilla verticillata biomass. Data obtained in batch experiments indicate that H. verticillata is an excellent biosorbent for Cd. Cd was rapidly adsorbed and such adsorption reached equilibrium within 20 min. The initial pH of the solution affected Cd sorption efficiency. Results obtained from the other batch experiments conformed well to those obtained using the Langmuir model. The maximum adsorption capacity q(max) for H. verticillata was 15.0 mg/g for Cd. The breakthrough curve from the continuous flow studies shows that H. verticillata in the fixed-bed column is capable of decreasing Cd concentration from 10 to a value below the detection limit of 0.02 mg/l. The presence of Zn ions affected Cd biosorption. It can be concluded that H. verticillata is a good biosorbent for treating wastewater with a low concentration of Cd contaminants.

Toxicity and bioaccumulation of cadmium and lead in Salvinia cucullata.

The toxicity and accumulation of heavy metals, cadmium (Cd) and lead (Pb) in aquatic fern, Salvinia cucullata were studied. Plants were cultured in Hoagland's medium which was supplemented with 0.5,1,2, and 4 mg/l of Cd and 5, 10, and 40 mg/l of Pb and were separately harvested after 2,4,6, and 8 days. The toxicity symptoms of Cd and Pb to S. cucullata showed chlorosis on leaves. There were significant derceases in the relative growth, biomass productivity and total chlorophyll content when the exposure time and concentration were increased. The accumulation study showed the significant increases of both metals when the exposure time and concentration were increased. The roots of S. cucullata had higher Cd and Pb contents than leaves suggesting that the metals were bound to the root cells and were partially transported to the leaves.

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.

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.

Molecular cloning and characterization of a glutathione S-transferase encoding gene from Opisthorchis viverrini.

An adult stage Opisthorchis viverrini cDNA library was constructed and screened for abundant transcripts. One of the isolated cDNAs was found by sequence comparison to encode a glutathione S-transferase (GST) and was further analyzed for RNA expression, encoded protein function, tissue distribution and cross-reactivity of the encoded protein with other trematode protein counterparts. The cDNA has a size of 893 bp and encodes a GST of 213 amino acids length (OV28GST). The most closely-related GST of OV28GST among those published for trematodes is a 28 kDa GST of Clonorchis sinensis as shown by multiple sequence alignment and phylogenetic analysis. Northern analysis of total RNA with a gene-specific probe revealed a 900 nucleotide OV28GST transcriptional product in the adult parasite. Through RNA in situ hybridization OV28GST RNA was detected in the parenchymal cells of adult parasites. This result was confirmed by immunolocalization of OV28GST with an antiserum generated in a mouse against bacterially-produced recombinant OV28GST. Both, purified recombinant and purified native OV28GST were resolved as 28 kDa proteins by SDS-PAGE. Using the anti-recOV28GST antiserum, no or only weak cross-reactivity was observed in an immunoblot of crude worm extracts against the GSTs of Schistosoma mansoni, S. japonicum, S. mekongi, Eurytrema spp. and Fasciola gigantica. The enzyme activity of the purified recombinant OV28GST was verified by a standard 1-chloro-2, 4-dinitrobenzene (CDNB) based activity assay. The present results of our molecular analysis of OV28GST should be helpful in the ongoing development of diagnostic applications for opisthorchiasis viverrini.

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 microm), 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.

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 (0.31 +/- 0.14 microm). This was in contrast with the larger size of intracellular particles of Se (2.37 +/- 1.84 microm) or W (1.75 +/- 1.34 microm). 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.