Bioavailability and extraction of heavy metals from contaminated soil by Atriplex halimus

Pot experiments were performed to evaluate the phytoremediation capacity of plants of Atriplex halimus grown in contaminated mine soils and to investigate the effects of organic amendments on the metal bioavailability and uptake of these metals by plants. Soil samples collected from abandoned mine sites north of Madrid (Spain) were mixed with 0, 30 and 60 Mg ha⁻¹ of two organic amendments, with different pH and nutrients content: pine-bark compost and horse- and sheep-manure compost. The increasing soil organic matter content and pH by the application of manure amendment reduced metal bioavailability in soil stabilising them. The proportion of Cu in the most bioavailable fractions (sum of the water-soluble, exchangeable, acid-soluble and Fe–Mn oxides fractions) decreased with the addition of 60 Mg ha⁻¹ of manure from 62% to 52% in one of the soils studied and from 50% to 30% in the other. This amendment also reduced Zn proportion in water-soluble and exchangeable fractions from 17% to 13% in one of the soils. Manure decreased metal concentrations in shoots of A. halimus, from 97 to 35 mg kg⁻¹ of Cu, from 211 to 98 mg kg⁻¹ of Zn and from 1.4 to 0.6 mg kg⁻¹ of Cd. In these treatments there was a higher plant growth due to the lower metal toxicity and the improvement of nutrients content in soil. This higher growth resulted in a higher total metal accumulation in plant biomass and therefore in a greater amount of metals removed from soil, so manure could be useful for phytoextraction purposes. This amendment increased metal accumulation in shoots from 37 to 138 mg pot⁻¹ of Cu, from 299 to 445 mg pot⁻¹ of Zn and from 1.8 to 3.7 mg pot⁻¹ of Cd. Pine bark amendment did not significantly alter metal availability and its uptake by plants. Plants of A. halimus managed to reduce total Zn concentration in one of the soils from 146 to 130 mg kg⁻¹, but its phytoextraction capacity was insufficient to remediate contaminated soils in the short-to-medium term. However, A. halimus could be, in combination with manure amendment, appropriate for the phytostabilization of metals in mine soils.     

Evaluation of phosphorus distribution and bioavailability in sediments of a subtropical wetland reserve in southeast China

The phosphorus (P) fractions and bioavailable P in the sediments from the Quanzhou Bay Estuarine Wetland Nature Reserve were investigated using chemical extraction methods for the first time to study the distribution and bioavailability of P in the reserve sediments. A hypothesis was presented suggesting that the bioavailable P in the sediments could be evaluated using the P fractions. The total phosphorus (TP), inorganic phosphorus (IP), organic phosphorus (OP), non-apatite phosphorus (NAIP), and apatite phosphorus (AP) contents in the sediments were in the ranges of 303.87–761.59 mg kg⁻¹, 201.22–577.66 mg kg⁻¹, 75.83–179.16 mg kg⁻¹, 28.86–277.90 mg kg⁻¹, and 127.36–289.94 mg kg⁻¹, respectively. The water soluble phosphorus (WSP), readily desorbable phosphorus (RDP), algal available phosphorus (AAP), and NaHCO₃ extractable phosphorus (Olsen-P) contents in the sediments were in the ranges of 0.58–357.17 mg kg⁻¹, 80.77–586.75 mg kg⁻¹, 1.09–24.12 mg kg⁻¹, and 54.96–676.82 mg kg⁻¹, respectively. The correlation analysis results showed that the NAIP was the major component of the bioavailable P and that the impact of the AP on the bioavailable phosphorus may be minimal. Due to the low TP content in the sediments of the Quanzhou Bay Estuarine Wetland Nature Reserve, the potential pollution risks of P in the sediments may not be very high. The results also show that the bioavailable P concentrations in the sediments of the Quanzhou Bay Estuarine Wetland Nature Reserve could not be evaluated by measuring the P fractions and that the hypothesis was untenable.     

Trace amounts of nickel in belowground rhizomes of Vaccinium myrtillus L. decrease anthocyanin concentrations in aerial shoots without water stress

Nickel (Ni) may impair plant water balance through detrimental effects on the belowground level. Bilberry (Vaccinium myrtillus L.) plants were grown in a mesic heath forest-type soil and subjected to Ni sulphate (NiSO₄·6H₂O) concentrations of 0, 10, 50, 100 and 500 mg m⁻² during an entire growing season in northern Finland (65°N). Biomass of belowground rhizomes, and tissue water content (TWC) and anthocyanin concentrations of aerial shoots were determined from mature plants in order to study rhizospheric Ni stress, and its possible long-distance effects on aerial shoots. As the major proportion of biomass of bilberry is invested in belowground parts, it was hypothesised that Ni-induced rhizospheric disturbance causes water stress in aerial shoots and increases their anthocyanin concentrations for osmotic regulation. Uptake of Ni from the soil to the rhizome and aerial shoots was measured with X-ray fluorescence spectrometry. Ni concentrations in the soil and rhizome exhibited a dose–response relationship, but the concentrations in the rhizome were about 10-fold lower (<3 mg Ni kg⁻¹) than those in the soil (<30 mg Ni kg⁻¹). Translocation of Ni from the rhizome to aerial shoots did not occur, as Ni concentrations in shoots remained at 1 mg Ni kg⁻¹. Although Ni concentrations in the rhizome were below the threshold values of Ni toxicity (i.e. 10–50 mg Ni kg⁻¹), Ni decreased the rhizome biomass. Anthocyanins decreased in aerial shoots along with the Ni accumulation in the rhizome, while TWC was unaffected. The result suggests that anthocyanins are not involved in osmotic regulation under Ni stress, since anthocyanins in aerial shoots responded to the Ni concentrations in the rhizome despite the lack of water stress.     

Stress responses and specific metal exclusion on mine soils based on germination and growth studies by Australian golden wattle

We reported the Australian golden wattle as a copper stabilizer in abandoned copper mine soils earlier. Here we investigate to confirm this plant’s suitability to grow on metal contaminated mine soils based on stress indication. The seeds of Acacia pycnantha collected from mining area were germinated after heat and no heat treatment on two types of irrigation. The daily irrigated and heat treated seeds gave up to 85% germination on sandy soil. The A. pycnantha was grown under greenhouse condition in six different soils collected from abandoned copper mine at Kapunda in South Australia. Among the six soil samples, soil-1 with the highest copper concentration produced 2.05 mmol g⁻¹ tissue of proline. Proline expression was prominent in more saline soils (1, 5 and 6) having electrical conductivity (EC) 1184, 1364 and 1256 μS, respectively. Chlorophyll a, b and carotenoid levels in plants showed a gradually decreasing trend in all the soils as experiment progressed. The plants grown on soil sample-1, containing 4083 ± 103 mg kg⁻¹ of copper resulted in 18 ± 2 mg kg⁻¹ accumulation in its leaf. The calcium accumulation was significant up to 11648 ± 1209 mg kg⁻¹ in leaf. Although pore water samples showed higher Cu concentration in soils, an increased mobility of arsenic and lead was observed in all the soil samples. Our experiment points out the need for proper monitoring of revegetation processes to avoid revegetation and reclamation failure.     

Assessment of Lemna gibba L. (duckweed) as a potential ecological indicator for contaminated aquatic ecosystem by boron mine effluent

Duckweeds, as a group, are important early warning indicators for the assessment of contaminated ecosystems due to their propensity to accumulate pollutants. In the present study, we investigated the potential use of Lemna gibba L. (Lemnaceae) as an ecological indicator for boron (B) mine effluent containing B concentration above 10 mg l⁻¹. For this purpose, L. gibba fronds were grown for 7 days in simulated water contaminated with B mine effluent. The important note is that this study was carried out in Kırka (Eskişehir, Turkey) B reserve area, which is the largest borax reserve in all over the world, under natural climatic conditions in the field. The results demonstrated that accumulations of B by L. gibba gradually increased based on the initial B concentrations (10, 25, 50, 100, and 150 mg l⁻¹) of the mine effluent. B concentration in the dry weight of the plant reached 639 mg kg⁻¹ when the minimum initial dosage (10 mg l⁻¹) was applied and 2711 mg kg⁻¹ when the maximum initial dosage (150 mg l⁻¹) was applied during the study. However, significant reductions in their relative growth rates occurred in 50, 100 and 150 mg l⁻¹ initial B concentrations. Results suggest that 25 mg l⁻¹ B concentration in water seemed to be a sensitive endpoint for L. gibba that could be used as a critical bioindicator level of B contaminated water. Following our data, we also constructed a simple growth model under the climatic conditions in this region of Turkey, but in instructive as a worldwide model. L. gibba is, therefore, suggested to be able to use as both an indicator and a phytoremediation tool because of its high accumulation capacity for B contaminated water.     

Selenium uptake,speciation and stressed response of Nicotiana tabacum L.

The research on the function and mechanism of selenium (Se) is of great significance for the development of Se-enriched agricultural products. In this paper, uptake, speciation distribution, the effects on the flue-cured tobacco growth and antioxidant system of Se at different levels (0–22.2 mg Se kg⁻¹) were studied through a pot experiment, aiming to clarify flue-cured tobacco's response to Se stress and the relationship between Se speciation and antioxidant system. The results showed that the leaf area and number, the biomass and the chlorophyll content reached the maximum at 4.4 mg kg⁻¹ of Se treatment. Selenium at low levels (≤4.4 mg kg⁻¹) stimulated the growth of flue-cured tobacco by elevating the capability of antioxidant stress and reducing the malondialdehyde (MDA) content to 0.6–0.8 times of that of the control. However, high Se levels (≥11.1 mg kg⁻¹) depressed the capability of antioxidant stress and raised the MDA content to 1.5-fold of that of the control, and meanwhile the biomass of the aboveground parts and underground parts declined notably. The Se content in different parts of flue-cured tobacco significantly increased with the growth of Se levels. The range of Se content in roots, leaves and stems at 2.2–22.2 mg kg⁻¹ of Se treatment were 16.7–58.6 mg kg⁻¹, 2.6–37.3 mg kg⁻¹ and 2.2–10.3 mg kg⁻¹, respectively. According to the detection of different Se speciation, only selenocysteine (SeCys) was detectable in leaves at 2.2 mg kg⁻¹ Se treatment; SeCys, selenite [Se(IV)]and selenate [Se(VI)] were detected in flue-cured tobacco leaves at Se treatment (≥4.4 mg kg⁻¹), which accounted for 4.6–10%, 9–18.7% and 71–86% respectively; SeCys, selenomethionine (SeMet) and Se(IV) were detected in roots, and organic selenium(66–84%) was the main Se species at Se ≤ 11.1 mg kg⁻¹ treatment; four Se species [SeCys, SeMet, Se(IV) and Se(VI)] were detected in flue-cured tobacco roots, and the main Se species was inorganic Se (60%) at 22.2 mg kg⁻¹ Se treatment. That was to say, the percentage of organic Se species (SeCys and SeMet in flue-cured tobacco leaves and root) declined, whereas the ratio of inorganic Se species [Se(IV) and Se(VI)] increased with the growth of Se levels. The correlation analysis showed that the superoxide dismutase (SOD) activity as well as the glutathione (GSH) and MDA contents were positively correlated with the Se(IV) and Se(VI) contents at P < 0.01 and excessive inorganic Se might destruct the reactive oxygen species (ROS) balance and enhance the MDA content, thus causing damage to the plant growth. In a word, the present study suggested that the ratio of inorganic Se [Se(IV) and Se(VI)] was closely related with the growth and the antioxidant capacity of flue-cured tobacco and the excessive application of Se led to the higher proportion of inorganic Se and poorer antioxidant capacity, which ultimately inhibited the growth of flue-cured tobacco.     

In vivo antimicrobial evaluation of an alanine-rich peptide derived from Pleuronectes americanus

In several organisms, the first barrier against microbial infections consists of antimicrobial peptides (AMPs) which are molecules that act as components of the innate immune system. Recent studies have demonstrated that AMPs can perform various functions in different tissues or physiological conditions. In this view, this study was carried out in order to evaluate the multifunctional activity in vivo of an alanine-rich peptide, known as Pa-MAP, derived from the polar fish Pleuronectes americanus. Pa-MAP was evaluated in intraperitoneally infected mice with a sub-lethal concentration of Escherichia coli at standard concentrations of 1 and 5 mg kg⁻¹. At both concentrations, Pa-MAPs exhibited an ability to prevent E. coli infection and increase mice survival, similar to the result observed in mice treated with ampicillin at 2 mg kg⁻¹. In addition, mice were monitored for weight loss. The results showed that mice treated with Pa-MAPs at 1 mg kg⁻¹ gained 0.8% of body weight during the 72 h of experiment. The same was observed with Pa-MAP at 5 mg kg⁻¹, which had a gain of 0.5% in body weight during the treatment. Mice treated with ampicillin at 2 mg kg⁻¹ show a significant weight loss of 5.6% of body weight. The untreated group exhibited a 5.5% loss of body weight. The immunomodulatory effects were also evaluated by the quantification of IL-10, IL-12, TNF-α, IFN-γ and nitric oxide cytokines in serum, but no immunomodulatory activity was observed. Data presented here suggest that Pa-MAP should be used as a novel antibiotic against infection control.     

Response of miscanthus to toxic cadmium applications during the period of maximum growth

Plants of miscanthus were grown in a Cd-free solution up to 1 month before heading and then were exposed to 0, 0.75, 1.5, 2.25 and 3 mg l⁻¹ cadmium for 36 days. All cadmium levels were toxic to miscanthus. Growth response was not dose-dependent and two toxicity thresholds were identified: one between 0 and 0.75 mg l⁻¹ Cd, the other between 2.25 and 3 mg l⁻¹ Cd. The former caused a biomass decrease by about 50%, whereas the latter completely inhibited growth and disrupted the mechanisms that restricted Cd translocation to the shoot. Growth of the aerial part was affected by cadmium more than that of the hypogeal one. Cadmium did not change the N concentration of different plant parts, but markedly reduced the N uptake of the plant, the N net uptake rate (NUR) and the N net translocation rate (NTR) from the rhizome to the aerial part. These two indexes equalled zero when plants ceased to grow. Otherwise, the Cd-NUR increased with Cd supply and the Cd-NTR from rhizome to aerial part showed the highest increment when plants did not grow at all. This suggests different uptake pathways for the two elements, active for nitrogen and passive for cadmium. The Cd concentration and the Cd content markedly increased with all Cd levels, following the order roots ≫ rhizome > culms > leaves. The Cd concentration and the Cd content of aerial organs increased with Cd supply, but increments were highest between 2.25 and 3 mg l⁻¹ Cd. The highest Cd concentrations were recorded in plants grown with 3 mg l⁻¹ Cd and were 41 and 122 mg kg⁻¹, respectively, for the aerial and the hypogeal plant parts. The hypogeal plant part retained most of the cadmium taken up from solution, accounting for approximately 87% of total plant cadmium with the three lower Cd levels, and for 73% with the highest one. The maximum Cd content of the entire plant was achieved with the two higher Cd levels and was approximately 4.7 mg, while the Cd content of the aerial part was highest with 3 mg l⁻¹ Cd (1.2 mg Cd per plant) and that of the hypogeal one with 2.25 mg l⁻¹ Cd (4 mg Cd per plant). The highest aerial content achieved in this experiment was 10-fold that obtained in a previous research when small-sized plants were exposed to the same Cd level.     

Response of native grasses and Cicer arietinum to soil polluted with mining wastes: Implications for the management of land adjacent to mine sites

Mine tailings are an environmental problem in Southern Spain because wind and water erosion of bare surfaces results in the dispersal of toxic metals over nearby urban or agricultural areas. Revegetation with tolerant native species may reduce this risk. We grew two grasses, Lygeum spartum and Piptatherum miliaceum, and the crop species Cicer arietinum (chickpea) under controlled conditions in pots containing a mine tailings mixed into non-polluted soil to give treatments of 0%, 25%, 50%, 75% and 100% mine tailings. We tested a neutral (pH 7.4) mine tailings which contained high concentrations of Cd, Cu, Pb and Zn. Water-extractable metal concentrations increased in proportion to the amount of tailings added. The biomass of the two grasses decreased in proportion to the rate of neutral mine-tailing addition, while the biomass of C. arietinum only decreased in relation to the control treatment. Neutron radiography revealed that root development of C. arietinum was perturbed in soil amended with the neutral tailings compared to those of the control treatment, despite a lack of toxicity symptoms in the shoots. In all treatments and for all metals, the plants accumulated higher concentrations in the roots than in shoots. The highest concentrations occurred in the roots of P. miliaceum (2500 mg kg^?1 Pb, 146 mg kg^?1 Cd, 185 mg kg^?1 Cu, 2700 mg kg^?1 Zn). C. arietinum seeds had normal concentrations of Zn (70–90 mg kg^?1) and Cu (6–9 mg kg^?1). However, the Cd concentration in this species was ～1 mg kg^?1 in the seeds and 14.5 mg kg^?1 in shoots. Consumption of these plant species by cattle and wild fauna may present a risk of toxic metals entering the food chain.     

Lead,zinc, cadmium hyperaccumulation and growth stimulation in Arabis paniculata Franch

Metal hyperaccumulation is of great interest in recent years because of its potential application for phytoremediation of heavy metal contaminated soils. In this study, a field survey and a hydroponic experiment were conducted to study the accumulation characteristics of lead (Pb), zinc (Zn) and cadmium (Cd) in Arabis paniculata Franch., which was found in Yunnan Province, China. The field survey showed that the wild population of A. paniculata was hyper-tolerant to extremely high concentrations of Pb, Zn and Cd, and could accumulate in shoots an average level of 2300 mg kg^?1 dry weight (DW) Pb, 20,800 mg kg^?1 Zn and 434 mg kg^?1 Cd, with their translocation factors (TFs) all above one. Under the hydroponic culture, stimulatory effects of Pb, Zn and Cd on shoot dry biomass were noted from 24 to 193 μM Pb, 9 to 178 μM Cd and all Zn supply levels in nutrient solution, while the effects were not obvious in the roots. Chlorophyll concentrations in Pb, Zn and Cd treatments showed an inverted U-shaped pattern, consistent with the change of plant biomass. Pb, Zn and Cd concentrations in the shoots and roots increased sharply with increasing Pb, Zn and Cd supply levels. They reached > 1000 mg kg^?1 Pb, 10,000 mg kg^?1 Zn and 100 mg kg^?1 Cd DW in the 24 μM Pb, 1223 μM Zn and 9 μM Cd treatment, respectively, in which the plants grew healthy and did not show any symptoms of phytotoxicity. The TFs of Zn were basically higher than one and the amount of Zn taken by shoots ranged from 78.7 to 90.4% of the total Zn. However, the TFs of Pb and Cd were well below one, and 55.0–67.5% of total Pb and 57.8–83.5% of total Cd was accumulated in the shoots. These results indicate that A. paniculata has a strong ability to tolerate and hyperaccumulate Pb, Zn and Cd. Meanwhile, suitable levels of Pb, Zn and Cd could stimulate the biomass production and chlorophyll concentrations of A. paniculata. Thus, it provides a new plant material for understanding the mechanisms of stimulatory effect and co-hyperaccumulation of multiple heavy metals.     

The relationship between Pseudo-nitzschia (Peragallo) and domoic acid in Scottish shellfish

The diatom genus Pseudo-nitzschia (Peragallo) associated with the production of domoic acid (DA), the toxin reposnsible for amnesic shellfish poisoning, is abundant in Scottish waters. A two year study examined the relationship between Pseudo-nitzschia cells in the water column and DA concentration in blue mussels (Mytilus edulis) at two sites, and king scallops (Pecten maximus) at one site. The rate of DA uptake and depuration differed greatly between the two species with M. edulis whole tissue accumulating and depurating 7 μg g⁻¹ (now expressed as mg kg⁻¹) per week. In contrast, it took 12 weeks for DA to depurate from P. maximus gonad tissue from a concentration of 68 μg g⁻¹ (now mg kg⁻¹) to <20 μg g⁻¹ (now mg kg^‐1). The DA depuration rate from P. maximus whole tissue was <5% per week during both years of the study. Correlations between the Pseudo-nitzschia cell densities and toxin concentrations were weak to moderate for M. edulis and weak for P. maximus. Seasonal diversity on a species level was observed within the Pseudo-nitzschia genus at both sites with more DA toxicity associated with summer/autumn Pseudo-nitzschia blooms when P. australis was observed in phytoplankton samples. This study reveals the marked difference in DA uptake and depuration in two shellfish species of commercial importance in Scotland. The use of these shellfish species to act as a proxy for DA in the environment still requires investigation.     

Use of lincomycin to control respiratory infections in lambs: Effects on health and production

The efficacy of lincomycin to control respiratory infections in lambs was assessed in two trials. In trial I, 72 lambs with active mycoplasmal pneumonia were allocated as follows: lambs in group T2 were treated with lincomycin (5 mg kg⁻¹ bodyweight, intramuscularly) twice 2 days apart, those in group T3 with lincomycin (5 mg kg⁻¹ bodyweight, intramuscularly) thrice with 2-day intervals, those in group O with oxytetracycline (20 mg kg⁻¹ bodyweight, intramuscularly) twice 4 days apart and those in group C were controls. In trial II, 48 25–30-day-old clinically healthy lambs were allocated as follows: lambs in group P2 received two injections of lincomycin (5 mg kg⁻¹ bodyweight, intramuscularly) when 30- and 60-day-old, lambs in group P1/30 received one injection of lincomycin (5 mg kg⁻¹ bodyweight, intramuscularly) when 30-day-old, lambs in group P1/60 received one injection of lincomycin (5 mg kg⁻¹ bodyweight, intramuscularly) when 60-day-old and lambs in group C were controls. In trial I, treatment with lincomycin was associated with improved clinical scores; clinical cure rate 42 days after treatment was 87%, 100%, 87% and 0% for group T2, T3, O and C, respectively (P < 0.001); treated lambs produced 18.5% (T2) or 26.5% (T3) heavier carcass than controls; no lung lesions were seen in group T3 lambs, whilst they were evident in 22% of group T2 or group O lambs and in 72% of control lambs; microorganisms were isolated from lung tissue samples of 5 group C and 1 group O lambs. In trial II, administration of lincomycin was associated with smaller clinical scores; prevalence rate of respiratory disorders at the end of the trial was 17%, 42%, 42% and 58% for group P2, P1/30, P1/60 and C, respectively (P < 0.01); treated lambs were >4.5% heavier than controls; lung lesions were recorded in 1 group P2, 2 group P1/30 and group P1/60 and 5 group C lambs; microorganisms were isolated from 1 group P2, 3 group P1/30, 2 group P1/60 and 5 group C lambs. It is concluded that administration of lincomycin is effective for the treatment and the prevention of mycoplasmal atypical pneumonia in lambs.     

Brachythecium rutabulum and Betula pendula as bioindicators of heavy metal pollution around a chlor-alkali plant in Poland

Chlor-alkali plants are known to be major sources of Hg emissions into the air. Therefore level of this metal in their surrounding must be carefully controlled. The aim of this work was to study the impact of the chlor-alkali industry in Brzeg Dolny (SW Poland) on the length of the vegetative short shoots of the pollution tolerant Betula pendula using the concentrations of Cd, Co, Cr, Cu, Fe, Hg, Mn, Ni, Pb and Zn in the ubiquitous, terrestrial moss Brachythecium rutabulum as the pollution indicator of the environment. This investigation showed up to 14 mg kg⁻¹ elevated concentrations of Hg in mosses from sites the most close (0–500 m) to the chlor-alkali factory. This concentration decreased with increasing distance from the factory. Two and half km away from the factory the Hg concentration falls to values of 0.1–0.2 mg kg⁻¹ being still higher than background concentrations of 0.03–0.04 mg kg⁻¹. Decreasing concentrations of Co, Cr, Fe and Ni were also correlated with increasing distance from the plant. The results indicate that B. rutabulum may be a suitable ecological indicator of metal pollution by chlor-alkali industry. Higher concentration of accumulated metals by this moss corresponds with longer vegetative short shoots of B. pendula. Vegetative short shoots may be used as bioindicators of metal pollution where mosses are absent. This study demonstrates the importance of controlling the emissions of chlor-alkali industry especially if situated in the midst of densely populated areas with potential risks to the inhabitants.     

Protocorm culture of Dendrobium candidum in balloon type bubble bioreactors

Protocorm cultures of Dendrobium candidum were established in balloon type bubble bioreactors using Murashige and Skoog (MS) medium with 0.5 mg l⁻¹ α-naphthaleneacetic acid (NAA), 2.5% (w/v) sucrose, 5:25 mM NH₄:NO₃ and 1% (v/v) banana homogenate for the production of biomass and bioactive compounds. In 3 l bioreactor containing 2 l medium, a maximum protocorm biomass (21.0 g l⁻¹ dry biomass) and also optimum quantities of total polysaccharides (389.3 mg g⁻¹ DW), coumarins (18.0 mg g⁻¹ DW), polyphenolics (11.9 mg g⁻¹ DW), and flavonoids (4.5 mg g⁻¹ DW) were achieved after 7 weeks of culture. Based on these studies, 5 and 10 l bioreactor cultures were established to harvest 80 g and 160 g dry biomass. In 10 l bioreactors, the protocorms grown were accumulated with optimal levels of polysaccharides (424.1 mg g⁻¹ DW), coumarins (15.8 mg g⁻¹ DW), polyphenols (9.03 mg g⁻¹ DW) and flavonoids (4.7 mg g⁻¹ DW). The bioreactor technology developed here will be useful for the production of important bioactive compounds from D. candidum.     

Reducing nutrient loss from farms through silvopastoral practices in coarse-textured soils of Florida,USA

Trees integrated into the range- and pasturelands of Florida could remove nutrients from deeper soil profiles that would otherwise be transported to water bodies and cause pollution. Soil nitrogen (N) and phosphorus (P) concentrations were monitored in three pastures: a treeless pasture of bahiagrass (Paspalum notatum); a pasture of bahiagrass under 20-year-old slash pine (Pinus elliotti) trees (silvopasture); and a pasture of native vegetation under pine trees (native silvopasture). Soil analysis from 10 profiles within each pasture showed that P concentrations were higher in treeless pasture (mean: 9.11 mg kg⁻¹ in the surface to 0.23 mg kg⁻¹ at 1.0 m depth) compared to silvopastures (mean: 2.51 and 0.087 mg kg⁻¹, respectively), and ammonium–N and nitrate–N concentrations were higher in the surface horizon of treeless pasture. The more extensive rooting zones of the combined stand of tree + forage may have caused higher nutrient uptake from silvopastures than treeless system. Further, compared to treeless system, soils under silvopasture showed higher P storage capacity. The results suggest that, compared to treeless pasture, silvopastoral association enhances nutrient retention in the system and thus reduces chances for nutrient transport to surface water. The study reflects the scope for applying ecological-engineering and ecosystem-restoration principles to silvopastoral-system design.     

Investigation on the properties and kinetics of glucose-fed aerobic granular sludge

Aerobic granulation is a process in which suspended biomass aggregate and form discrete well-defined granules in aerobic systems. To investigate the properties and kinetics of aerobic granular sludge, aerobic granules were cultivated with glucose synthetic wastewater in a series of sequencing batch reactors (SBR). The spherical shaped granules were observed on 8th day with the mean diameter of 0.1 mm. With the organic loading rate (OLR) being increased to 4.0 g COD L⁻¹ d⁻¹, aerobic granules grew matured with spherical shape. The size of granules ranged from 1.2 to 1.8 mm, and the corresponding settling velocity of individual granule was 24.2–36.4 m h⁻¹. The oxygen utilization rate (OUR) of mature granules was 41.90 g O₂ kg MLSS⁻¹ h⁻¹, which was two times higher than that of activated sludge (18.32 g O₂ kg MLSS⁻¹ h⁻¹). The experimental data indicated that the substrate utilization and biomass growth kinetics generally followed Monod's kinetics model. The corresponding kinetic coefficients of k (maximum specific substrate utilization rate), K_s (half velocity coefficient), Y (growth yield coefficient) and K_d (decay coefficient) were determined as follows, k_c = 23.65 d⁻¹, K_c = 3367.05 mg L⁻¹, K_N = 0.038 d⁻¹, K_N = 29.65 mg L⁻¹, Y = 0.1927–0.2022 mg MMLS (mg COD)⁻¹ and K_d = 0.00845–0.0135 d⁻¹, respectively. Those properties of aerobic granules made aerobic granules system had a short setup period, high substrate utilization rate and low sludge production.     

The phytoremediation potential for strontium of indigenous plants growing in a mining area

This study investigated the distribution and accumulation of strontium (Sr) in the shoots and roots of Euphorbia macroclada (EU), Verbascum cheiranthifolium (VR), and Astragalus gummifer (AS), with respect to their potential use in phytoremediation. Plant samples and their associated soils were collected from the arid and semi-arid Keban mining area and were analyzed inductively by ICP-MS for Sr. Mean Sr values in the shoots, roots and soil were, respectively, 453, 243 and 398 mg kg^?1 for E. macroclada; 149, 106 and 398 mg kg^?1 for V. cheiranthifolium; and 278, 223 and 469 mg kg^?1 for A. gummifer. The enrichment factors for root (ECR) and shoot (ECS) of these plants were lower than 1 or close to 1, except for the shoot of E. macroclada. The mean translocation factors (TLF) of these plants were higher than 1 and 2.08 for E. macroclada, 1.47 for V. cheiranthifolium, 1.18 for A. gummifer. It thus appeared that the shoots of these plants can be an efficient bioaccumulator plant for Sr and it can be used in cleaning or rehabilitating of the contaminated soil and areas by Sr because of their high translocation factors.     

Evaluation of zinc tolerance and accumulation potential of the coastal shrub Limoniastrum monopetalum (L.) Boiss.

The coastal shrub Limoniastrum monopetalum is capable of growth in soil containing extremely high concentrations of heavy metals. A greenhouse experiment was conducted in order to investigate the effects of a range of Zn concentrations (0–130 mmol l⁻¹) on growth and photosynthetic performance, by measuring relative growth rate, total leaf area, plant height, gas exchange, chlorophyll fluorescence parameters and photosynthetic pigment concentrations. We also determined the total zinc, nitrogen, phosphorus, sulphur, calcium, magnesium, sodium, potassium, iron and copper concentrations in the plant tissues. The study species demonstrated hypertolerance to Zn stress, since survival was recorded with leaf concentrations of up to 1700 mg Zn kg⁻¹ dry mass when treated with 130 mmol Zn l⁻¹. L. monopetalum exhibited little overall effects on photosynthetic function at Zn levels of up to 90 mmol l⁻¹. At greater external Zn concentration, plant growth was negatively affected, due in all probability to the recorded decline in net photosynthetic rate, which may be linked to the adverse effect of the metal on photosynthetic electron transport. Growth parameters were virtually unaffected by leaf tissue concentrations as high as 1400 mg Zn kg⁻¹ dry mass thus indicating that this species could play an important role in the phytoremediation of Zn-polluted areas.     

Improvement of succinate production by overexpression of a cyanobacterial carbonic anhydrase in Escherichia coli

Succinate fermentation was investigated in Escherichia coli strains overexpressing cyanobacterium Anabaena sp. 7120 ecaA gene encoding carbonic anhydrase (CA). In strain BL21 (DE3) bearing ecaA, the activity of CA was 21.8 U mg⁻¹ protein, whereas non-detectable CA activity was observed in the control strain. Meanwhile, the activity of phosphoenolpyruvate carboxylase (PEPC) increased from 0.2 U mg⁻¹ protein to 1.13 U mg⁻¹ protein. The recombinant bearing ecaA reached a succinate yield of 0.39 mol mol⁻¹ glucose at the end of the fermentation. It was 2.1-fold higher than that of control strain which was just 0.19 mol mol⁻¹ glucose. EcaA gene was also introduced into E. coli DC1515, which was deficient in glucose phosphotransferase, lactate dehydrogenase and pyruvate:formate lyase. Succinate yield can be further increased to 1.26 mol mol⁻¹ glucose. It could be concluded that the enhancement of the supply of HCO₃⁻ in vivo by ecaA overexpression is an effective strategy for the improvement of succinate production in E. coli.     

Effect of the polycyclic aromatic hydrocarbon phenanthrene on root exudation of Sorghum bicolor (L.) Moench

The objective of this work was to study the effect of two concentrations (10 and 100 mg kg⁻¹) of phenanthrene, a ubiquitous polycyclic aromatic hydrocarbon (PAH), on root exudation of the remediating plant Sorghum bicolor (L.) Moench under controlled conditions in a pot experiment. It was found that the phenanthrene concentration of 10 mg kg⁻¹ did not cause significant effects on plant survival and growth but had little stimulating effect on carbohydrate exudation. The contamination with phenanthrene at 100 mg kg⁻¹ inhibited accumulation of plant shoot and root biomass, decreasing the carboxylic acid, carbohydrate, and amino acid amounts released by sorghum root into the rhizosphere. However, root exudation per unit of root surface was not changed significantly with increasing phenanthrene concentration. There were no differences in qualitative composition of root exudates under the influence of PAH were found. The observed alterations in the ratio between the main root-exuded components are assumed to manifest adaptive alterations occurring in the plant as a response to pollutant stress. The activity of three oxidoreductases (oxidase, peroxidase, and tyrosinase) released by sorghum roots was clearly progressive to the increasing phenanthrene concentration in the substrate. Under the influence of phenanthrene, the population of phenanthrene-degrading microorganisms in sorghum root zone increased, and their share in the total number of culturable heterotrophs increased as well. The main promotional factor was the pollutant; however, the stimulating effect of the plant root exudates was also involved. The increased pollutant-degrading microbial population and activity of the extracellular root enzymes are presumed to be important for the rhizodegradation of PAH.