An ecological dose-response model approach to short- and long-term effects of heavy metals on arylsulphatase activity in soil

Summary The aim of this study was to provide data to evaluate the short- and long-term effects of heavy metals on arylsulphatase activity in five soils. The effects are fitted on a logistic dose-response model and are presented graphically as the ecological dose (heavy metal concentration corresponding to 50% inhibition; ED₅₀) and ecological dose range (heavy metal concentration range corresponding to 10–90% inhibition; EDR). In 7 out of 22 comparable soil-metal combinations the ED₅₀ decreased significantly over 6 weeks to 18 months of incubation and in two cases the ED₅₀ increased. Toxicity (defined as ED₅₀) was highest in sand and sandy loam and lowest in sandy peat. Cd toxicity in sand, silty loam, and clay varied from 1.08 to 9.04 mmol kg^-1. Both Cr and Ni toxicity varied strongly and decreased with time in some soils while increasing in others. The Cu toxicity ranged from 4.51 to 2 mmol kg^-1 in sand and silty loam, respectively, but remained fairly constant over time. Pb was the least toxic element (14.5 to 59.9 mmol kg^-1). The toxicity of Zn ranged from 5.73 to 148 mmol kg^-1 in sand and sandy peat, respectively. At critical concentrations set by the Dutch Soil Protection Act, Cr, Cu, Ni, and Zn inhibited arylsulphatase by 53, 35, 48 and 97%, respectively.     

Short- and long-term effects of heavy metals on phosphatase activity in soils: An ecological dose-response model approach

Summary The aim of this study was to provide manageable data to help establish permissible limits for the pollution of soil by heavy metals. Therefore the short-and long-term effects of heavy metal pollution on phosphatase activity was studied in five different soil types. The results are presented graphically as logistic dose-response curves. It was possible to construct a curve for sand and silty loam soil but it was more difficult to establish a curve for sandy loam and clay soil and nearly impossible (except for Cu) for peat. The toxicity of the various metals can be compared on the basis of mmol values. In clay soils, for Cd, Cr, Cu, and Zn, the 50% effective ecological dose (ED₅₀) values were comparable (approximately 45 mmol kg^–1), but the ED₁₀ values were very different, at 7.4, 41.4, 15.1, and 0.55, respectively. At the ED₅₀ value, toxicity did not decrease with time and, in sandy soils, was approximately 2.6 mmol kg ^–1 dry soil for Cd, Cu, and Zn. In four out of five soils, the Cd toxicity was higher 1.5 years after the addition of heavy metal salts than after 6 weeks. Toxicity was least in the sandy loam, silty loam, and clay soil, and varied in general between 12 and 88 mmol kg^–1. In setting limits, the criteria selected (no-effect level, ED₁₀ or ED₅₀) determine the concentration and also the toxicity of the sequence. It is suggested that the data presented here could be very useful in helping to set permissible limits for heavy metal soil pollution.     

The Ecological Dose of Nickel in a Semiarid Soil Amended with Sewage Sludge Related to the Unamended Soil

Nickel pollution may affect microbial communitydevelopment and its activity in soil and, therefore, itsfertility, while organic amendment may affect Ni mobilityand bioavailability. The aim of this research was toascertain the extent to which the addition of sewage sludgeto soil may affect Ni toxicity and to establish theecological dose of this heavy metal. Unamended and amendedsoils with sewage sludge were spiked with differentconcentrations of Ni (0–8000 mg kg^-1) and then incubatedfor 3 hours, 12 days and 40 days. Soil dehydrogenaseactivity, ATP content, microbial respiration and microbialbiomass C were measured in the samples containing differentNi concentrations. A mathematical model which describe theinhibition of these soil parameters were used to calculatethe ecological dose of Ni: ED₅₀. Soil microbialbiomass C measurements of all treatments better fitted themodel (r² = 0.95–0.69 with p < 0.05) than the otherparameters studied; but the ED₅₀ values predicted by the modelhad a high coefficient of variation. For both ATP contentand microbial biomass C, the ED₅₀ values calculatedfor the amended soil were higher than for the unamendedsoil and so that the effect of Ni toxicity on theseparameters of microbial activity may be considered lower inthe former. In this study, an increase in ED₅₀ valueswith the time elapsed was also observed.     

Evaluation of isotopic dilution method for measuring N2 fixation in Azolla: Comparison with other methods 1

An isotopic dilution method that overcomes the drawbacks of commonly used methods for measuring N₂ fixation by aquatic N‐fixers such as Azolla pinnata‐Anabaena azollae association (Azolla) is presented. The method was compared with ¹⁵N₂ gas (while maintaining CO₂) and the difference methods of measuring N₂ fixation. The isotopic dilution method was used for two conditions: a. For ¹⁵N‐free growth medium, Azolla was pre‐enriched with ¹⁵N, and N₂ fixation was determined by measuring the dilution of ¹⁵N in the tissue. b. For the growth medium containing N, N₂ fixation was determined by providing ¹⁵N enriched ammonium sulfate in the growth medium and measuring ¹⁵N to ¹⁴N ratio in the tissue. An airtight chamber, necessary for ¹⁵N₂ gas and acetylene reduction methods, was not representative of the growing environment of Azolla. Temperature in the airtight chamber was far from uniform and CO₂ was rapidly depleted. The isotopic dilution method is simpler, relatively inexpensive, subject to fewer errors and applicable to more diverse conditions, and yet was as accurate as ¹⁵N₂‐gas method.     

Porous tubing for use in monitoring soil CO2 concentrations

The composition of the soil atmosphere is an indicator of biological processes, and soil CO₂ gradients have been used to estimate CO₂ efflux from the surface. Soil atmosphere samplers, constructed with gas-permeable materials, have been used to quantify soil CO₂ concentrations. The type of material used can influence the perceived real-time concentrations of CO₂ in the soil. Previous works have not directly compared different types of materials under the same conditions. The objective of this study was to determine the diffusion coefficient (D) and time of 95% equilibrium (t_eq) of CO₂ through several materials, and to evaluate the effect of long-term soil burial (183 days) on diffusion characteristics. Materials tested included silicone, expanded Teflon (ePTFE), and ultra high molecular weight polyethylene (PE) tubing. The D of each material was determined using a closed-loop system consisting of a CO₂-enriched (7800 ppm) chamber, a CO₂ analyzer and an inner tube (experimental tubing) placed inside the chamber. Air was re-circulated through the inner tube, and as CO₂ diffused from the chamber into the tubing, the analyzer recorded the increase in concentration. The silicone tubes had values of D ranging from 8.64 to 5.80×10⁻⁶ cm² s⁻¹ with corresponding t_eq between 3.9 and 9.7 h. Diffusion coefficients of the ePTFE (1.25×10⁻⁴ cm² s⁻¹) and PE (7.70×10⁻⁴ cm² s⁻¹) materials were 2 orders of magnitude greater, with t_eq<6 min. Exposure to the soil environment for 183 days did not visibly deteriorate the materials or significantly affect the D or t_eq values. Use of the ePTFE or PE materials, over the silicone materials, may allow for better characterization of dynamic CO₂ concentrations in the soil based on the greater D and lesser t_eq values of these materials.     

Evolution of CO2 from soil incubated with dieldrin-C and the action of soil bacteria on labelled dieldrin

Arable soil containing 10 ppm of dieldrin-¹⁴C uniformly labelled in its chlorinated ring released 0·30 and 1.86 per cent of the activity as ¹⁴CO₂ from sterile and non-sterile samples respectively during 7 weeks incubation. In a second experiment with percolated or aerated soil samples containing 50 ppm dieldrin-¹⁴C with or without glucose about 0·30 per cent was lost as ¹⁴CO₂. The different experimental conditions only influenced the time pattern of ¹⁴CO₂ evolution. About half of 177 bacterial strains isolated from the same soil produced water-soluble dieldrin metabolites in culture. From 14 selected strains the three most active strains (Nocardia, Corynebacterium and aMicrococcus sp.) were incubated for 5 weeks with 0·4 ppm dieldrin-¹⁴C and released 0·06–0·11 per cent recoverable as BaCO₃ in aerated culture and 0·14–0·2 per cent in stationary culture.     

Nitrogen fixation in association with the root systems of goldenrods (Solidago L.)

The roots of seven species of goldenrod (Asteraceae: Solidago) were assayed for associative nitrogen fixation. We used the acetylene reduction method to measure N₂ fixation rates in sampling jars that contained roots of sampled plants and soil. The rate of C₂H₂ reduction in jars with S. rigida (12.9 ± 1.2 nmol h⁻¹, mean ± SE) and S. canadensis (6.0 ± 2.0 nmol h⁻¹) was significantly higher than in soil-only control jars. There was measurable C₂H₂ reduction in jars of the other five species, but the rates were not significantly different from controls. The rates of C₂H₂ reduction per root dry weight and per total plant dry weight also were higher for S. rigida and S. canadensis than for the other species.     

Effects of 2,4-d on nitrogen fixation and carbon dioxide evolution in a soil microcosm

Two concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) 1.7 kg ha^?1 and 3.4 kg ha^?1 were applied to oats (Avena sativa L. ‘Orbit’) grown in terrestrial microcosms in a sandy soil. Carbon dioxide evolution and non-symbiotic N₂ fixation (C₂H₂ reduction) were measured weekly. On day 70 of the study, 2,4-D was applied a second time at the same application rates and soil CO₂ evolution and N₂ fixation were measured more frequently. Soil CO₂ evolution over 24 h period was significantly decreased by 40 to 50% at both application rates of 2,4-D. This response lasted less than 7 days. Nitrogen fixation was unaffected by 2,4-D except for an unexplained decrease observed in the 1.7 kg ha^?1 treatment 35 days after 2,4-D application. This effect was not observed on the following sampling date. The second application of 2,4-D failed to produce any significant change in soil CO₂ evolution or N₂ fixation. From these studies we conclude soil microbial populations either degraded or became acclimated to 2,4-D as a result of the initial application and that 2,4-D has no significant adverse effect on N₂ fixation or soil CO₂ evolution.     

Dynamics of C natural abundance in wood decomposing fungi and their ecophysiological implications

Factors that affect the δ¹³C values of fungi need to be analyzed for the progress of isotope-based studies of food-chain or organic matter dynamics in soils. To analyze the factors that control δ¹³C values of the fungal body, basidiomycete and ascomycete species were grown on a beechwood substrate (six species) and in glucose medium (nine species), and the δ¹³C value of produced fungal body was compared to that of the carbon source. The ¹³C enrichment (Δδ¹³C) in the fungal aggregates compared to the decomposed wood varied from 1.2 to 6.3‰ among six species. In the glucose substrate experiment, the degree of ¹³C enrichment in the hyphal mat was relatively small and varied from −0.1 to 2.8‰ among nine basidiomycetes species depending on their growth stage. Calculated δ¹³C values of the respired CO₂ were lower than those of the hyphal mat, organic metabolites and the glucose used. The degree of ¹³C enrichment was affected by fungal species, substrate and growth stage. Fungal internal metabolic processes are the plausible mechanism for the observed isotopic discrimination between fungal bodies and substrates. Especially, dark fixation of ambient CO₂ and kinetic isotope fractionation during assimilation and dissimilation reactions could well explain Δδ¹³C dynamics in our experiments. Through the analysis of field Δδ¹³C, we could know undisturbed fungal status about starvation, aeration and type of decomposition.     

Effect of elevated atmospheric CO2 and temperature on disease severity of Fusarium oxysporum f.sp. lactucae on lettuce plants

Climate change effects on Fusarium oxysporum f.sp. lactucae (FOL) on lettuce plants grown under simulated climate change were studied. An artificial growing substrate was infested with FOL to reach a final concentration of 1 × 10⁴ CFU g⁻¹ of substrate. A non infested tank was used as control.Subsequently, 32 pots (2 l each) were prepared from the infested tank and other 32 pots were prepared from the non infested tank (control). Lettuce plants were then transplanted into the pots and grown in phytotrons under four simulated environmental conditions: (1) 800 ppm CO₂, 22–26 °C; (2) 800 ppm CO₂, 18–22 °C; (3) 400 ppm CO₂, 22–26 °C and (4) 400 ppm CO₂, 18–22 °C. Substrate samples were collected from each phytotron at 7, 14, 21 and 28 days after transplanting. Plate counts, enzymatic assays, and polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) analyses were performed to evaluate the effect of climate change on the microbial population. The abundance of Fusarium spp. and the severity of Fusarium wilt of lettuce varied significantly as a consequence of increased temperature (22–26 °C). Increased CO₂ levels showed no effect on the severity of Fusarium wilt of lettuce and on the abundance of Fusarium spp. On the other hand, the total bacterial abundance was reduced at elevated CO₂ concentration (800 ppm). PCR-DGGE fingerprints of the ascomycete community obtained from DNA directly extracted from infested substrate samples did not change as a consequence of elevated temperature and CO₂.Enzymatic activities were not affected by the elevated CO₂ level. Our study indicates that the CO₂ concentration used in our experiment had no detectable impact on Fusarium wilt of lettuce. Only temperature influenced all observed parameters, but did not affect the fungal species diversity. Other factors, such as nutrient limitation and the effect of plant species needs further study.     

Elevated CO<Subscript>2</Subscript> alters the abundance but not the structure of diazotrophic community in the rhizosphere of soybean grown in a Mollisol

The effect of elevated CO₂ (eCO₂) on rhizospheric diazotrophic community in cropland has little been studied, although eCO₂ facilitates nodulation and N₂ fixation in legumes. In this study, four soybean cultivars (Xiaohuangjin, Suinong 8, Suinong 14, and Heinong 45) were grown in Mollisols for 65 days under ambient CO₂ (aCO₂) (390 ppm) or eCO₂ (550 ppm). Quantitative PCR and Illumina MiSeq sequencing targeting the nifH gene that reflects the composition of diazotrophic community were determined. Elevated CO₂ significantly increased the abundance of nifH gene copies in the rhizospheres of the Suinong 8 and Heinong 45 cultivars, but not in the Suinong 14 and Xiaohuangjin cultivars. The nifH abundance correlated negatively with nodule density (p?≤?0.01) but positively with nodule size (p?≤?0.01). Elevated CO₂ did not significantly alter the composition of diazotrophic community, nor shift dominant bacterial operational taxonomic units (OTUs). These results indicated that eCO₂ stimulated the growth but did not alter the community composition of diazotrophs in the rhizosphere of soybean, which depended on cultivar and might contribute to nodulation responses to eCO₂.     

Microbial activity and bacterial composition of H2-treated soils with net CO2 fixation

The effects of H₂ gas treatment of an agricultural soil cultivated previously with a mixture of clover (Trifolium pratense) and alfalfa (Medicago sativa) on CO₂ dynamics and microbial activity and composition were analyzed. The H₂ emission rate of 250 nmol H₂ g⁻¹ soil h⁻¹ was similar to the upper limit of estimated H₂ amounts emitted from N₂ fixing nodules into the surrounding soil ([Dong, Z., Layzell, D.B., 2001. H₂ oxidation, O₂ uptake and CO₂ fixation in hydrogen treated soil. Plant and Soil 229, 1-12.]). After 1 week of H₂ supply to soil samples simultaneously with H₂ uptake net CO₂ production declined continuously and this finally led to a net CO₂ fixation rate in the H₂-treated soil of 8 nmol CO₂ g⁻¹ soil h⁻¹. The time course of H₂ uptake and CO₂ fixation in the soils corresponded with an increase in microbial activity and biomass of the H₂-treated soil determined by microcalorimetric measurements, fluorescence in situ hybridization analysis (FISH) and DNA staining (DAPI). Shifts in the bacterial community structure caused by the supply of H₂ were recorded. While the H₂ treatment stimulated β-and γ-subclasses of Proteobacteria, it had no significant effect on α-Proteobacteria. In addition, FISH-detectable bacteria of the Cytophaga-Flavobacterium-Bacteroides phylum increased in numbers.     

Using the Caenorhabditis elegans soil toxicity test to identify factors affecting toxicity of four metal ions in intact soil

A previously developed soil toxicity test for rapidly determining the toxicity of chemicals to the soil-dwelling nematode Caenorhabditis elegans (Donkin and Dusenbery, 1993) was used to measure the toxicity of four metals (Zn²⁺, Cd²⁺, Cu²⁺, and Pb²⁺) added to four soils common to the southeastern United States. Nematode survival after a 24-hour exposure in the presence of a bacterial food source was assessed. All soils reduced the toxicity of most metal ions compared to solutions without soil. Pb was the most strongly affected, while Cd toxicity was not much influenced by the soils. Correlations between the LC₅₀S and various soil or metal characteristics were determined. No significant correlation was found between LC₅₀s and many soil characteristics commonly cited as having large effects on soil bioavailability of metals. Although sample size was limited, the indication was that bioavailability of metals to nematodes is determined by a complex array of many interacting soil, as well as metal, properties. Comparison of the relative mobilities of these ions in other soils with the relative toxicity measured here suggests that mobility may be a good predictor of toxicity. The C. elegans soil toxicity test is shown to be as sensitive and more rapid than the commonly used earthworm soil toxicity test.     

Lead accumulation from non-saline and saline environment by Tamarix smyrnensis Bunge

Tamarix smyrnensis plants were exposed to Pb(NO₃)₂, with and without addition of salt into soil for 10 weeks. Salt as NaCl was added to water for watering in concentrations 0.5% and 3%. Subsequently, lead uptake was quantified in leaves and roots of the plants by ICP-AES spectrometry. In addition, the influence of Pb on photosynthesis and other physiological parameters was also examined by means of biomass production and shoot length determination. Plant appearance was observed also. The influence of Pb presence in the soil on chlorophyll content was examined. In order to quantify the fate of Pb in the plant, metal excretion by salt glands was investigated as well. It was found that the roots were the main accumulation site of Pb in all plants under all treatments irrespective of salt concentration in soil. Low salinity in soil did not affect Pb accumulation in roots but at high salt concentration Pb accumulation decreased. The opposite was observed in the leaves where more Pb was accumulated at high salt concentration in soil. The toxic effect of Pb was visible only in the group treated with Pb without addition of NaCl into water for watering. The visible toxicity symptoms were connected only with high salinity. No excretion of Pb by salt glands was observed.     

Inhibitory effects of the total and water-soluble concentrations of nine different metals on the dehydrogenase activity of a loess soil

 This study focuses on a comparison of the microbial toxicity of nine metals, including As as a metalloid and two species of Cr. A loess soil [Ap horizon, clay 15.2%, organic C 1.12%, pH(CaCl₂) 7.02] was spiked with 8–12 geometrically increasing doses of the metals. The dehydrogenase assay (2-p-iodophenyl-3-p-nitrophenyl-5-phenyltetrazoliumchloride method) was combined with sorption and solubility experiments. The resulting dose-response curves and sorption isotherms were used to derive total doses that caused definite percentage inhibitions [i.e. effective doses (ED) causing a 10–90% reduction in dehydrogenase activity (dha)] as well as the corresponding toxic solution concentrations causing the same reductions in dha (i.e. effective concentrations; EC₁₀–EC₉₀). Based on total doses, the toxicity decreased in the following order with ED₅₀ values (mg kg^–1) given in brackets: Hg (2.0)>Cu (35)>Cr(VI) (71)>Cr(III) (75)>Cd (90)>Ni (100)>Zn (115)>As (168)>Co (582)>Pb (652). With regard to solution concentrations, toxicity decreased in the order (EC₅₀ in mg l^–1): Hg (0.003)>Pb (0.04)>Cu (0.05)>Cd (0.14)>Zn (0.19)>Cr(III) (0.62)>Ni (0.69)>Co (30.6)>As (55.5)>Cr(VI) (78.1). The retention of the metals by the soil differed strongly. Pb, Cu, and Hg exhibited the highest and Ni, As, and Cr(VI) the lowest sorption constants (Freundlich K values: 2455, 724, 348, 93, 13, and 0.06 mg kg^–1, respectively). The sorptivity of the metals and their microbial toxicity in the aqueous phase were characteristically related: metals with a strong toxic action in the soil solution were adsorbed by the soil to a high degree and vice versa. Therefore, especially for metals with a high inherent toxicity, sorption is an effective way of immobilizing them and temporarily detoxifying soil. Received: 2 July 1998     

Plant biomass influences rhizosphere priming effects on soil organic matter decomposition in two differently managed soils

We used a continuous labeling method of naturally ¹³C-depleted CO₂ in a growth chamber to test for rhizosphere effects on soil organic matter (SOM) decomposition. Two C3 plant species, soybean (Glycine max) and sunflower (Helianthus annus), were grown in two previously differently managed soils, an organically farmed soil and a soil from an annual grassland. We maintained a constant atmospheric CO₂ concentration at 400±5 ppm and δ¹³C signature at −24.4‰ by regulating the flow of naturally ¹³C-depleted CO₂ and CO₂-free air into the growth chamber, which allowed us to separate new plant-derived CO₂-C from original soil-derived CO₂-C in soil respiration. Rhizosphere priming effects on SOM decomposition, i.e., differences in soil-derived CO₂-C between planted and non-planted treatments, were significantly different between the two soils, but not between the two plant species. Soil-derived CO₂-C efflux in the organically farmed soil increased up to 61% compared to the no-plant control, while the annual grassland soil showed a negligible increase (up to 5% increase), despite an overall larger efflux of soil-derived CO₂-C and total soil C content. Differences in rhizosphere priming effects on SOM decomposition between the two soils could be largely explained by differences in plant biomass, and in particular leaf biomass, explaining 49% and 74% of the variation in primed soil C among soils and plant species, respectively. Nitrogen uptake rates by soybean and sunflower was relatively high compared to soil C respiration and associated N mineralization, while inorganic N pools were significantly depleted in the organic farm soil by the end of the experiment. Despite relatively large increases in SOM decomposition caused by rhizosphere effects in the organic farm soil, the fast-growing soybean and sunflower plants gained little extra N from the increase in SOM decomposition caused by rhizosphere effects. We conclude that rhizosphere priming effects of annual plants on SOM decomposition are largely driven by plant biomass, especially in soils of high fertility that can sustain high plant productivity.     

The ecological dose value (ED50) for assessing Cd toxicity on ATP content and dehydrogenase and urease activities of soil

Two soils of contrasting texture, organic matter content and pH were treated with CdSO₄ solutions to give a Cd concentration range of 0–4000 mg kg⁻¹ soil. The content of ATP and dehydrogenase and urease activities of soils were assayed after 3 h, and 7 and 28 days of Cd contamination. The relative ED₅₀ values were calculated by two kinetic models (model 1 and model 2) used by Speir et al. (1995) and by the sigmoidal dose–response model (model 3) employed by Haanstra et al. (1985). Model 1 was the most successful in calculating the ED₅₀ values for the ATP content, urease and dehydrogenase activities when both soils were contaminated by Cd. Similar ED₅₀ values were predicted by model 1 (describing the full inhibition) and model 3 only when the correlation coefficients r² were higher than 0.9. The ED₅₀ values of ATP calculated by model 1 were markedly higher than those calculated by model 2 (describing partial inhibition) when both models gave correlation coefficients higher than 0.9. This behavior was due to the high asymptote values obtained using model 2. According to model 2, some of the enzyme activities responsible for the ATP synthesis were probably not inhibited at the highest Cd concentrations. The inhibitory effect of Cd on the ATP content and both enzymatic activities was lower in the Castelporziano soil, which had the highest total organic carbon content.     

Evaluation of an optimal extraction method for measuring d-ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) in agricultural soils and its association with soil microbial CO2 assimilation

Assimilating atmospheric carbon (C) into terrestrial ecosystems is recognized as a primary measure to mitigate global warming. Ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) is the dominant enzyme by which terrestrial autotrophic bacteria and plants fix CO₂. To investigate the possibility of using RubisCO activity as an indicator of microbial CO₂ fixation potential, a valid and efficient method for extracting soil proteins is needed. We examined three methods commonly used for total soil protein extraction. A simple sonication method for extracting soil protein was more efficient than bead beating or freeze–thaw methods. Total soil protein, RubisCO activity, and microbial fixation of CO₂ in different agricultural soils were quantified in an incubation experiment using ¹⁴C-CO₂ as a tracer. The soil samples showed significant differences in protein content and RubisCO activity, defined as nmol CO₂ fixed g⁻¹ soil min⁻¹. RubisCO activities ranged from 10.68 to 68.07 nmol CO₂ kg⁻¹ soil min⁻¹, which were closely related to the abundance of cbbL genes (r = 0.900, P = 0.0140) and the rates of microbial CO₂ assimilation (r = 0.949, P = 0.0038). This suggests that RubisCO activity can be used as an indicator of soil microbial assimilation of atmospheric CO₂.     

Comparing the Tolerance Limits of Selected Bacterial and Protozoan Species to Vanadium in Wastewater Systems

This study compared the tolerance limits of selected bacterial (Bacillus licheniformis, Brevibacillus lactosporus and Pseudomonas putida) and protozoan (Aspidisca, Trachelophyllum and Peranema) species to V⁵⁺ in wastewater systems. The isolates were exposed to various concentrations of V⁵⁺ (from 10 to 240?ppm), and their tolerance limits to this heavy metal were assessed at different temperatures (25, 30, 35 and 40°C) and pHs (4, 6, 7, 8 and 10) for 5?days. Chemical oxygen demand (COD), dissolved oxygen (DO) and die-off rate of the isolates were measured using standard methods. The results indicated that test isolates were tolerant to V⁵⁺, with a gradual decrease in their colony/cell counts when V⁵⁺ concentration gradually increased. Bacterial species were found to be more significantly tolerant (MIC: 110?C230?ppm?V⁵⁺) to V⁵⁺ than protozoan species which showed an earlier total inhibition/die-off rate (100%) at 60?C100?ppm?V⁵⁺ (MIC) (p?<?0.001). P. putida was the most tolerant bacterial species (MIC: 230?ppm?V⁵⁺) and Aspidisca sp. the most sensitive protozoan species (MIC: 60?ppm?V⁵⁺). An increase in COD and DO removal was observed throughout the experimental period. The highest COD increase (up to 237.11%) and DO removal (almost 100%) were observed in mixed liquor inoculated with P. putida after exposure to 10?ppm?V⁵⁺. Changes in pH and temperature affected the tolerance limits of all isolates. This study suggests the use of these tolerant bacterial and protozoan species in the bioremediation of V⁵⁺ from domestic and industrial wastewater under the control of pH and temperature.     

Acute and sub-chronic toxicity of lead to the early life stages of smallmouth bass (Micropterus Dolomieui)

The early life stages of smallmouth bass (Micropterus dolomieui) were exposed to Pb in acute (96 hr) and sub-chronic (90 day) bioassays (water hardness = 152 mg L^?1 as CaCO₃). After 96-hr static exposures at nominal Pb concentrations up to 15.9 mg L^?1, eggs and sac fry showed no increased mortality over that in controls. Swim-up fry (96-hr LC₅₀ = 2.8 mg Pb L^?1) were more sensitive to Pb exposure than were fingerlings (96-hr LC₅₀ of 29.0 mg Pb L^?1 ). The relation between dissolved Pb and mortality was non-significant for either swim-up fry or fingerlings. Fingerlings were exposed to Pb concentrations as high as 405 μg L^?1 for 90 day to evaluate effects on substrate selection, locomotor activity, hematology, and weight. Dark or light substrate selection (cover-seeking) and locomotor activity, weight and hemoglobin concentration in the blood were not significantly altered by any treatment. Hematocrit and leucocrit varied significantly but not in relation to Pb levels. Sub-chronic Pb exposure did not appear to represent a threat to smallmouth bass in waters of medium hardness and above-neutral pH (7.1 to 7.9).