Effects of Amendments on Growth and Uptake of Cd and Zn by Wetland Plants, Typha angustifolia and Colocasia esculenta from Contaminated Sediments.

A pot study was conducted to compare the effects of amendments (CaHPO4 and cow manure) on growth and uptake of Cd and Zn from contaminated sediments by two wetland plant species, Typha angustifolia and Colocasia esculenta. Contaminated sediments (Cd 33.2 mg kg(-1) and Zn 363 mg kg(-1)) were collected from Mae Tao basin, Mae Sot district, Tak province, Thailand. The experiment consisted of 4 treatments: control (uncontaminated sediment), Cd/Zn, Cd/Zn + 5% CaHPO4, and Cd/Zn + 10% cow manure. Plants were grown for 3 months in the greenhouse. The addition of CaHPO4 resulted in the highest relative growth rate (RGR) and highest Cd accumulation in both T. angustifolia and C. esculenta while the lowest RGR was found in C. esculenta grown in the cow manure treatment. Both plant species had higher concentrations of metals (Cd, Zn) in their belowground parts. None of the amendments affected Zn accumulation. C. esculenta exhibited the highest uptake of both Cd and Zn. The results clearly demonstrated the phytoremediation potential of C. esculenta and the enhancement of this potential by CaHPO4 amendment.

Phytoremediation potential of Cd and Zn by wetland plants, Colocasia esculenta L. Schott., Cyperus malaccensis Lam. and Typha angustifolia L. grown in hydroponics.

Cadmium and zinc phytoremediation potential of wetland plants, Colocasia esculenta, Cyperus malaccensis, and Typha angustifolia, was investigated. Plants were grown for 15 days in nutrient solutions containing various concentrations of Cd (0, 5, 10, 20, 50 mg l(-1)) and Zn (0, 10, 20, 50, 100 mg l(-1)). T angustifolia was tolerant to both metals as indicated by high RGR when grown in 50 mg I(-1) Cd and 100 mg I(-1) Zn solutions. All these plants accumulated more metals in their underground parts and > 100 mg kg(-1) in their aboveground with TF values < 1. Only C. esculenta could be considered a Zn hyperaccumulator because it could concentrate > 10,000 mg kg(-1) in its aboveground parts with TF > 1. T angustifolia exhibited highest biomass production and highest Cd and Zn uptake, confirming that this plant is a suitable candidate for treating of Cd contaminated soil/sediments.

Histopathological effects of contaminated sediments on golden apple snail (Pomacea canaliculata, Lamarck 1822).

Pomacea canaliculata were exposed experimentally to contaminated sediments from a tributary of the Mae Klong River, Thailand, for 3 months. The highest concentration of Cr, Zn and Fe accumulated in the digestive gland while the gill exhibited the highest concentration of Cu. In addition, histopathological changes (increased mucus vacuoles, loss of cilia, dilation of cells in the epithelial cells of digestive tract organs, and an increase in the number of dark granules in the digestive cells) were observed. The gill exhibited loss of cilia, wider hemolymph space, and degeneration of columnar epithelial cells.

Tolerance and accumulation of lead in Vetiveria zizanioides and its effect on oil production.

Experiments were conducted to evaluate lead tolerance and accumulation in vetiver grass Vetiveria zizanioides (L.), grown in hydroponics and a pot study and to examine the effect of lead on vetiver oil production. Elevated concentrations of lead decreased the length of shoots and roots of plants. However, vetiver grown in highly contaminated soils showed no apparent phytotoxicity symptoms. Lead concentrations in the shoots and roots of vetiver plants grown in hydroponics were up to 144 and 19530 mg kg(-1) and those grown in soil were 38 and 629 mg kg(-1), respectively. Lead had an effect on vetiver oil production and composition by stimulating oil yield and the number of its constituents. Oil yield ranged from 0.4-1.3%; the highest yields were found in plants grown in nutrient solution with 100 mg Pb l(-1) for 5 weeks (1.29%) and 7 weeks (1.22%). The number of total constituents of vetiver oil also varied between 47-143 compounds when lead was presentin the growth medium. The highest number (143) was found in plants grown in soil spiked with 1000 mg Pb kg(-1). The predominant compound was khusimol (10.7-18.1%) followed by (E)-isovalencenol (10.3-15.6%). Our results indicated that lead could increase the oil production of vetiver.

Genome-wide identification of genes involved in tolerance to various environmental stresses in Saccharomyces cerevisiae.

During fermentation, yeast cells are exposed to a number of stresses -- such as high alcohol concentration, high osmotic pressure, and temperature fluctuation - so some overlap of mechanisms involved in the response to these stresses has been suggested. To identify the genes required for tolerance to alcohol (ethanol, methanol, and 1-propanol), heat, osmotic stress, and oxidative stress, we performed genome-wide screening by using 4828 yeast deletion mutants. Our screens identified 95, 54, 125, 178, 42, and 30 deletion mutants sensitive to ethanol, methanol, 1-propanol, heat, NaCl, and H2O2, respectively. These deleted genes were then classified based on their cellular functions, and cross-sensitivities between stresses were determined. A large number of genes involved in vacuolar H(+)-ATPase (V-ATPase) function, cytoskeleton biogenesis, and cell wall integrity, were required for tolerance to alcohol, suggesting their protective role against alcohol stress. Our results revealed a partial overlap between genes required for alcohol tolerance and those required for thermotolerance. Genes involved in cell wall integrity and the actin cytoskeleton are required for both alcohol tolerance and thermotolerance, whereas the RNA polymerase II mediator complex seems to be specific to heat tolerance. However, no significant overlap of genes required for osmotic stress and oxidative stress with those required for other stresses was observed. Interestingly, although mitochondrial function is likely involved in tolerance to several stresses, it was found to be less important for thermotolerance. The genes identified in this study should be helpful for future research into the molecular mechanisms of stress response.

Residue analysis of organochlorine pesticides in the Mae Klong River of Central Thailand.

The most natural waterways in Thailand are utilized for conventional agricultures, e.g. irrigation and wastewater discharge. Concentrations of 20 organochlorine pesticides (OCPs) were measured in surface sediments collected from the tributaries of the Mae Klong River during 2003-2005. The sampling sites chosen were nearby the rice paddy which organochlorine pesticides were intensively used for crop cultivation. Solid-liquid extraction technique was used for sediment extraction, and gas chromatography with electron capture detector was applied for pesticides determination. Total concentrations of OCPs found were ranged from 4.12 to 214.91 microg g(-1) dw in the first year. In the second year, total residue concentrations in the surface sediments were in the range of 3.26-215.09 microg g(-1) dw. Concentrations of pesticides were greater in dry-season rice cultivation period in both sampling years, and heptachlor epoxide was the contaminant found at highest concentration in this study. In addition, the occurrence of some selected contaminants including DDT, DDD and DDE reflects the recent contamination in the study area. This demonstrates not only that the natural waterway was polluted and contained much pesticide residues, but it also illustrates the necessity of a continuous long-term monitoring of the affected environment.

Uptake and accumulation of cadmium, lead and zinc by Siam weed [Chromolaena odorata (L.) King & Robinson].

The Siam weed, Chromolaena odorata (L.) King & Robinson, Family Asteraceae, was found to be a new Pb hyperaccumulator by means of field surveys on Pb soil and hydroponic studies. Plants from field collection accumulated 1377 and 4236mgkg(-1) Pb in their shoots and roots, respectively, and could tolerate soil Pb concentrations up to 100000 mgkg(-1) with a translocation factor of 7.62. Very low concentrations of Cd and Zn were found in plants collected from the field. Under nutrient solution culture condition, C. odorata from the contaminated site (CS) and from non-contaminated site (NCS) grew normally with all three metals (Pb, Cd, Zn) supplied. However, the relative growth rates of all treated plants decreased with increased metal concentrations. The percentage uptakes of Pb, Cd, and Zn by C. odorata increased with increasing metal concentrations. Pb concentration in shoots and roots reached its highest values (1772.3 and 60655.7mgkg(-1), respectively) at a Pb supply level of 10mgl(-1). While the maximum concentrations of Cd (0.5mgl(-1)) in shoots and roots of C. odorata were 102.3 and 1440.9mgkg(-1), and the highest concentrations of Zn (20mgl(-1)) were 1876.0 and 7011.8mgkg(-1), respectively. The bioaccumulation coefficients of Pb and Cd were greater than 1000. These results confirm that C. odorata is a hyperaccumulator which grows rapidly, has substantial biomass, wide distribution and has a potential for the phytoremediation of metal contaminated soils.

Growth and lead accumulation by the grasses Vetiveria zizanioides and Thysanolaena maxima in lead-contaminated soil amended with pig manure and fertilizer: a glasshouse study.

Bo Ngam lead mine soils contain high concentrations of lead (up 1% total Pb) and low amounts of organic matter and major nutrients (N, P, K). A glasshouse study was conducted to compare growth performance, metal tolerance and metal uptake by two grasses, Thysanolaena maxima (Roxb.) O. Kuntze and four ecotypes of Vetiveria zizanioides (L.) Nash, syn. Chrysopogon zizanioides (L.) Roberty (three from Thailand: Surat Thani, Songkhla and Kamphaeng Phet, and one from Sri Lanka) and to study the effects of pig manure (20% and 40% w/w) and inorganic fertilizer (75 and 150 mg kg(-1)) amendments to this lead mine soil. The results showed that both T. maxima and V. zizanioides (Surat Thani and Songkhla) could tolerate high Pb concentrations in soil (10750 mg kg(-1)) and had very good growth performance. Application of pig manure increased electrical conductivity (EC) and reduced DTPA-extractable Pb concentration in the soils. Pig manure application improved the growth of vetiver, especially at 20%, application dosage. Vetiver had the highest biomass. T. maxima could not tolerate high EC values. The uptake by roots and transport of Pb to shoots of both species was reduced when soils were amended with pig manure. Application of inorganic fertilizer did not improve growth of vetiver but did improve that of T. maxima. Fertilizer application did not have any great influence on the Pb uptake in vetiver while T. maxima took up more Pb as a result of the fertilizer enhancing its biomass yield. Both species transported low Pb concentrations to shoots (8.3-179 mg kg(-1)) and accumulated higher concentrations in roots (107-911 mg kg(-1)). In summary, both species may be species well suited for phytostabilization in tropical lead mine areas.

Uptake and accumulation of lead by plants from the Bo Ngam lead mine area in Thailand.

A field survey of terrestrial plants growing on Bo Ngam lead mine area, Thailand, was conducted to identify species accumulating exceptionally high concentrations of lead. Plant and soil samples were collected from five areas. Lead concentrations in surface soil ranged from 325 to 142,400 mg/kg. The highest lead concentration in soil was found at the ore dressing plant area and lowest at a natural pond area. In different areas, the concentrations of lead in plants were different when comparing various study sites. A total of 48 plant species belonging to 14 families were collected from five sampling sites. Twenty-six plant species had lead concentrations more than 1000 mg/kg in their shoots. Three species (Microstegium ciliatum, Polygala umbonata, Spermacoce mauritiana) showed extremely high lead concentrations in their shoots (12,200-28,370 mg/kg) and roots (14,580-128,830 mg/kg).

Growth of mixed erythroid-granulocytic colonies in culture derived from bone marrow of patients with paroxysmal nocturnal hemoglobinuria without addition of exogenous stimulator.

Hematopoietic progenitor cell cultures were studied in 16 patients with paroxysmal nocturnal hemoglobinuria (PNH). Both erythroid and granulocyte-macrophage progenitors in the blood and bone marrow decreased despite hypercellular marrow. In eight out of 16 patients, mixed erythroid-granulocytic colonies were observed in the bone marrow cultures without erythropoietin and colony-stimulating factor. Removal of monocytes and T-lymphocytes before culture failed to abrogate spontaneous mixed-colony formation. Conditioned media prepared from blood and bone marrow of PNH patients could not promote the growth of mixed colonies in normal individuals. The autonomous, spontaneous mixed-colony formation in patients with PNH is an abnormality of PNH stem cells not described previously.

Towards practical cadmium phytoextraction with Noccaea caerulescens.

A series of field trials were conducted to investigate the potential of Noccaea caerulescens F.K. Mey [syn. Thlaspi caerulescens J &C Presl. (see Koch and Al-Shehbaz 2004)] populations (genotypes) derived from southern France to phytoextract localized Cd/Zn contamination in Thailand. Soil treatments included pH variation and fertilization level and application of fungicide. N. caerulescens populations were transplanted to the field plots three months after germination and harvested in May, prior to the onset of seasonal rains. During this period growth was rapid with shoot biomass ranging from 0.93-2.2 g plant(-1) (280-650 kg ha(-1)) DW. Shoot Cd and Zn concentrations for the four populations evaluated ranged from 460-600 and 2600-2900 mg kg(-1) DW respectively. Cadmium and Zn Translocation Factors (shoot/root) for the populations tested ranged from 0.91-1.0 and 1.7-2.1 and Bioaccumulation Factors ranged from 12-15 and 1.2-1.3. We conclude that optimizing the use of fungicidal sprays, acidic soil pH, planting density and increasing the effective cropping period will increase rates of Cd and Zn removal enough to facilitate practical Cd phytoextraction from rice paddy soils in Thailand.

Cadmium biosorption by Sphingomonas paucimobilis biomass.

Among microorganisms isolated in Bangkok, the gram-negative bacterium Sphingomonas paucimobilis exhibited the greatest cadmium tolerance. It was able to survive in the medium containing cadmium as high as 200 mg/l. However, concentrations of cadmium at 25-200 mg/l inhibited its growth. The biosorption properties for cadmium of this bacterial biomass and the effects of environmental factors (i.e., biosorbent type, initial pH and biosorbent concentration) on the cadmium biosorption were explored. The results showed that the cadmium removal capacity of living cells was markedly higher than that of nonliving cells. Cadmium biosorption by S. paucimobilis biomass was also affected by the initial pH and biosorbent concentration.

Kinetics of basic dye (methylene blue) biosorption by giant duckweed (Spirodela polyrrhiza).

Wastewater containing pigments and/or dyes can cause serious water pollution problems in the form of reduced light penetration and photosynthesis, and the toxicity from heavy metals associated with pigments and/or dyes. Laboratory investigations, of the potential use of dried Spirodela polyrrhiza biomass as an adsorbent for the removal of the basic dye methylene blue from aqueous solution were conducted. A series of experiments were undertaken in an agitated batch adsorber to assess the effect of the system variables, i.e. sorbent dosage, pH, and contact time. The results showed that as the amount of the dried S. polyrrhiza increased, the percentage of dye sorption increased accordingly. At pH 2.0 the sorption of dye was not favorable, while the sorption at other pHs (3.0-11.0) was remarkable. There was no significant difference in the dye concentration remaining when the pH was increased from 3.0 to 11.0. The dye removal time was influenced by the initial dye concentration, and the process followed the first-order rate kinetics. The rate constants for intraparticle diffusion were 1.00 and 3.27 mg/g/min1/2 for 300 and 500 mg/l of dye, respectively.

Phytoremediation potential of Spirulina (Arthrospira) platensis: biosorption and toxicity studies of cadmium.

This study examines the possibility of using Spirulina (Arthrospira) platensis TISTR 8217 to remove low concentrations of cadmium (less than 100 mg/l) from wastewater. The cyanobacteria were exposed to six different cadmium concentrations for 96 h, and the growth rate was determined using an optical density at 560 nm. The inhibiting concentration (IC50) was estimated using probit analysis. The IC50 at 24, 48, 72, and 96 h were 13.15, 16.68, 17.28, and 18.35 mg/l Cd, respectively. Cellular damage was studied under a light microscope and a transmission electron microscope. Swollen cells and fragmented filaments were observed. Cell injury increased with increasing concentrations of cadmium. Ultrastructural changes were observed in the algae exposed to cadmium concentrations both close to IC50 (14.68 mg/l) and at IC50 (18.35 mg/l). The alterations induced by cadmium were disintegration and disorganization of thylakoid membranes, presence of large intrathylakoidal space, increase of polyphosphate bodies, and cell lysis. In addition, the cadmium adsorption by algal cells was studied. Environmental factors were found to have an effect on biosorption. The uptake of cadmium was not affected by the temperature of the solution, but the sorption was pH dependent. The optimum pH for biosorption of algal cells was 7. The cadmium uptake process was rapid, with 78% of metal sorption completed within 5 min. The sorption data fit well to the Langmuir isotherm. The maximum adsorption capacity for S. platensis was 98.04 mg Cd per g biomass.

Histopathological alterations of white seabass, Lates calcarifer, in acute and subchronic cadmium exposure.

Histopathological alterations to white seabass, Lates calcarifer aged 3 months in acute and subchronic cadmium exposure were studied by light and scanning electron microscopy. The 96-h LC50 values of cadmium to L. calcarifer was found to be 20.12 +/- 0.61 mg/l and the maximum acceptable toxicant concentration (MATC) was 7.79 mg/l. Fish were exposed to 10 and 0.8 mg/l of Cd (as CdCl,H2O) for 96 h and 90 days, respectively. The study showed that gill lamellae and kidney tubules were the primary target organs for the acute toxic effect of cadmium while in the subchronic exposure, the toxic effect to gills was less than that of kidney and liver. Gill alterations included edema of the epithelial cells with the breakdown of pillar cell system, aneurisms with some ruptures, hypertrophy and hyperplasia of epithelial and chloride cells. The liver showed blood congestion in sinusoids and hydropic swelling of hepatocytes, vacuolation and dark granule accumulation. Lipid droplets and glycogen content were observed in hepatocytes at the second and third month of subchronic exposure. The kidney showed hydropic swelling of tubular cell vacuolation and numerous dark granule accumulation in many tubules. Tubular degeneration and necrosis were seen in some areas.

Investigation of Nematopsis spp. oocysts in 7 species of bivalves from Chonburi province, Gulf of Thailand.

This is the first detailed report of Nematopsis spp. in Thai bivalves. A monthly survey was conducted on 7 species of commercial bivalves from Chonburi province, on the eastern seaboard of Thailand, from November 2000 to November 2001 to investigate the prevalence of the apicomplexan parasite Nematopsis Schneider, 1892. Nematopsis spp. sporozoites were found in the cultivated bivalves Arcuatula arcuatula, Anadara granosa and Perna viridis as well as the locally harvested Paphia undulata. They were not found in Donax faba, Meretrix meretrix or Saccostrea cucullata. Using light microscopy, we were able to identiby 4 oocyst morphotypes of the gregarine Nematopsis spp. Prevalence of Nematopsis spp. during the 13 mo sampling period was highest in A. arcuatula (91.8%; n = 110) and lowest in A. granosa (59.2%; n = 130). The morphology of the oocysts differed between hosts, with an average (x +/- SD) length/width of 16.28 +/- 0.64/12.01 +/- 0.35 microm (n = 50) for A. arcuatula, 16.90 +/- 0.71/12.69 +/- 0.33 microm (n = 50) for A. granosa, 17.61 +/- 0.69/12.72 +/- 0.36 microm (n = 50) for P. viridis, and 11.21 +/- 0.62/8.55 +/- 0.52 microm (n = 50) for P. undulata. Identification of oocysts of these apicomplexan gregarines to species was not attempted. The prevalence of infection in relation to habitat and time of sampling is discussed.

Effects of cercarial concentration and length of exposure on the infection of mice by Schistosoma japonicum and S. mekongi cercariae.

The effects of cercarial concentration and length of exposure on the infection of mice by Schistosoma japonicum and S. mekongi were studied under laboratory conditions. For all cercarial concentrations of 10, 20 and 30, mice exposed for 1 min acquired little or no infection. Total worm recovery tended to rise with an increase either in cercarial concentration or exposure time. The highest infection rate and worm burden were obtained in mice exposed for 64 min to 30 cercariae per mouse. The present investigation suggested that exposures of 1 min or less carried little risk of infection at low cercarial concentration. There was a greater risk of infection in prolonged exposure to low cercarial concentration. Prolonged exposures to high cercarial concentration carried the greatest risk of high infection rate and worm burden.

Effects of physico-chemical factors on the infection of mice with Schistosoma japonicum and S. mekongi cercariae.

The three physico-chemical characteristics of water tested-pH, temperature and salinity were found to affect the infection of mice by cercariae of S. japonicum and S. mekongi. For both species of schistosomes, the range of optimal pH that showed highest infection and worm recovery rates was between 5.4-7.4 and decreased as the pH increased. Cercariae exposed to temperatures between 15 degrees-30 degrees C showed the highest infection and worm recovery rates. As temperature decreased or increased, the infection and worm recovery rates decreased. Sodium chloride concentration at low level (0.5-200 ppm) did not have great effect on the infection and worm recovery rates. The infection and worm recovery rates decreased as salinity increased. High sodium chloride concentration at 3200 ppm for S. mekongi cercariae and at 6400 ppm for S. japonicum cereariae inhibited cercarial penetration and no infection occurred.

Scanning electron microscopy of adult Schistosoma incognitum.

The integumental surfaces of adult Schistosoma incognitum were studied by scanning electron microscopy. The anterior part of the male which bears oral and ventral suckers is spongy in appearance due to the presence of ridges, while the middle and posterior parts are covered with numerous bosses with variable numbers of spines. Surfaces between bosses possess tegumental papillae. Bosses disappear at the edge of the gynecophoral canal which is composed of ridges. They are replaced by variable forms of papillae. The lining of the gynecophoral canal is composed of ridges and papillae, free of spines, with small number of sensory bulbs. The female tegument is relatively smooth and appears to be pitted with minute spines around the oral and ventral suckers. The posterior end of the body, in the vicinity of the excretory pore, is composed of prominent anteriorly directed spines with trabeculae and sensory bulbs.

Scanning and transmission electronmicroscopy of Mekong Schistosoma eggs and adults.

Eggs and adult males and females of Mekong Schistosoma were studied by scanning and transmission electronmicroscopy. The observation of the eggs by the scanning and light microscopy revealed fine shell fenestration and a prominent knoblike spine. There are marked differences between the surface structures of male and female as studied by scanning and transmission electronmicroscopy. The surface of the male schistosome is moderately rough while that of the female is relatively smooth. SEM reveals certain basic features such as spines in the oral sucker, minute spines and folds in the gynecophoral canal of the male, and general features of male and female tegumental surfaces. The observations of the cross sections of adult schistosomes by transmission technique revealed certain features such as spines or ridges, and mucin droplets on the surface, the smooth muscles lining the integument, the mucin-producing cells and numerous lipid droplets in the body of the Schistosoma.