Ecotoxicity of soils contaminated with industrial and domestic wastewater in western shenyang,China

Soil samples were collected from 7 sites in the up-, mid-and down-reach along and nearby the wastewater irrigation channel, western Shenyang of China. The concentrations of selected pollutants (mineral oil, PAHs - polycycle aromatic hydrocarbons and Cd) were determined by UV spectrometer, HPLC and AAS (atomic adsorption spectrometer) spectrometer, respectively. Toxicity effects of soils were evaluated by seedling emergence test with root length of wheat as the end-point and by earthworms test with the mortality rate and inhibition rates of body weight as endpoints. Results showed accumulation of pollutants for most soils with concentration of 200.2 mg.kg^?1～1600 mg.kg^?1 for mineral oil, 0.33 mg.kg^?1～1.81 mg.kg^?1 for Cd and 900.16 mg.kg^?1 ～ 2737.91 mg.kg^?1 for PAHs. The inhibition rates of root elongation were from ?20% up to 40 %, and mortality rates of earthworms ranged from 0%～40% from the exposure period of two weeks to eight weeks by sampling interval of two weeks, the inhibition rates of earthworm growth were from ?19.36% to 34.53%, showing effects of stimulation at 2 weeks to an increasing effects of inhibition at 4, 6 and 8 weeks, respectively. Mortality rates correlated with the loss of body weight of earthworms.This study indicated the potential risk of pollutants of environmental low content in soil by the determination of selected chemicals combined with toxicity indexes.     

Ecotoxicity of soils contaminated with industrial and domestic wastewater in western Shenyang, China

Soil samples were collected from 7 sites in the up-, mid- and down-reach along and nearby the wastewater irrigation channel, western Shenyang of China. The concentrations of selected pollutants (mineral oil, PAHs - polycycle aromatic hydrocarbons and Cd) were determined by UV spectrometer, HPLC and AAS (atomic adsorption spectrometer) spectrometer, respectively. Toxicity effects of soils were evaluated by seedling emergence test with root length of wheat as the end-point and by earthworms test with the mortality rate and inhibition rates of body weight as endpoints. Results showed accumulation of pollutants for most soils with concentration of 200.2 mg.kg-1 ～1600 mg.kg-1 for mineral oil, 0.33 mg.kg-1～1.81 mg.kg-1 for Cd and 900.16 mg.kg-1～ 2737.91 mg.kg-1 for PAHs. The inhibition rates of root elongation were from -20% up to 40 %, and mortality rates of earthworms ranged from 0%～40% from the exposure period of two weeks to eight weeks by sampling interval of two weeks, the inhibition rates of earthworm growth were from -19.36% to 34.53%, showing effects of stimulation at 2 weeks to an increasing effects of inhibition at 4, 6 and 8 weeks, respectively. Mortality rates correlated with the loss of body weight of earthworms.This study indicated the potential risk of pollutants of environmental low content in soil by the determination of selected chemicals combined with toxicity indexes.     

Initial results for urban metal distributions in house dusts of syracuse,New York,USA

A program of house dust sample collection and analysis has begun in Syracuse, New York, USA, in order to determine the feasibility of a geography-based exposure assessment for urban metals. The sampling program, and the protocols it employs, is described for two different types of wipe media, Ghost Wipes and Whatman Filters. Preliminary results show that strong spatial patterns of floor dust loading (mg dust per square foot) can be observed for data aggregated at a spatial scale of about 1600 m (∼2.5 km²). Floor dust metal concentrations were similar to those found in other urban environments, with some regional variation. The median floor dust Pb concentration was ∼108 mg · kg⁻¹ for this initial data set of ∼264 sampled residential locations, and varied from 50 to 1100 mg Pb · kg⁻¹.

     

Polycyclic Aromatic Hydrocarbon (PAH) Contamination in the Urban Topsoils of Shenyang,China

Polycyclic aromatic hydrocarbons (PAHs) in the surface urban soils of Shenyang in Northeastern China were investigated. The total concentration of the PAHs ranged from 0.09 to 8.35 mg kg^?1, with an average value of 1.51 ± 1.64 mg kg^?1. 3–5-ring PAHs accounted for 90% of total PAHs. The functional areas, such as the industrial regions (4.95 mg kg^?1) and main roads (1.56 mg kg^?1), as well as the administrative divisions, including the districts of Shenhe (1.49 mg kg^?1), Heping (2.08 mg kg^?1), and Tiexi (2.14 mg kg^?1), were heavily polluted by PAHs. The diagnostic ratios and principal component analysis (PCA) for PAHs indicate that the pollutants probably originated primarily from coal combustion and petroleum sources. The Nemerow composite index, used to assess environmental quality, shows that the soil samples were heavily polluted with PAHs, and although 52.8% of the soil sampling sites were safe, 47.2% of the soil sampling sites registered different grades of PAH pollution. The PAH contamination in Shenyang emphasizes the need for controlling fossil fuel combustion and traffic exhaust.     

Activated sludge under free ammonia treatment using gel immobilization technology for long-term partial nitrification with different initial biomass

To achieve stable partial nitrification, activated sludge from a wastewater treatment plant using free ammonia (FA) inhibition was immobilized in a polyvinyl alcohol carrier. After FA treatment at 16.44 mg L⁻¹ for 1 day, due to the increased growth rate gap between ammonium-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB), AOB enrichment and NOB inhibition were achieved within 12 days, with AOB and NOB accounting for 65.61 and 0.05%, respectively. Subsequently, with dissolved oxygen concentrations of 4−5 mg L⁻¹, pH of 7.6–7.8 and temperature of 25 ± 1 °C, the immobilized carrier made of activated sludge achieved more than 90% and more than 86% of nitrite accumulation rate at the influent ammonia concentration of 90−110 mg L⁻¹ and 35−50 mg L⁻¹, respectively. After 50 days operation, the NOB content was 0.10%, indicating the immobilized carrier provided favorable conditions for maintaining the low NOB content. Furthermore, due to the low NOB content in the inoculum and the oxygen-limited environment formed by the increase in the AOB numbers in the carrier, immobilized carrier with different initial biomass (1, 2.5 and 5%) can achieve stable partial nitrification.     

Assessment of germination and carnivorous activities of a nematode-trapping fungus Arthrobotrys dactyloides in fungistatic and fungicidal soil environment

Carnivorism is the ability of nematode-trapping fungi to trap and digest the nematodes by sophisticated devices called traps. Delivery of nematode-trapping fungi in soil for bio-control of pest nematodes often fails or gives inconsistent results. Possible reasons for failure could be the effect of soil fungistasis on germination of nematode-trapping fungi in soil environment, use of avirulent species and sensitivity of these fungi to fungicidal residues in soil. Exploitation of nematode-trapping fungi for nematode control demands that it be compatible with fungicides applied in soil or crops and proliferate in soil. This investigation represents is one of the first to evaluate the effect of fungicides on the nematode-trapping fungus Arthrobotrys dactyloides. A. dactyloides showed in vitro carnivorous potential against Meloidogyne incognita, Meloidogyne javanica, Meloidogyne graminicola, Helicotylenchus dihystera and Heterodera cajani. Conidia of A. dactyloides exposed to agricultural soils showed poor germination but formed conidial traps, which captured and killed the soil nematodes. Conidial traps, which trapped the nematodes, grew well in all soils after killing and nutrient absorption from nematode body. Soil amended with 20 mg ai kg⁻¹ of carbendazim and thiram, 30 mg ai kg⁻¹ of mancozeb, 50 mg ai kg⁻¹ of captan, and 100 mg ai kg⁻¹ of carboxin completely checked the conidial trap formation and nematode capturing. 30, 50 and 100 mg ai kg⁻¹ of metalaxyl adversely affected the conidial trap formation and nematode capturing in soil. Propiconazole inhibited 15.2% conidial trap formation up to 50 mg ai kg⁻¹ but caused 93.3% inhibition of conidial traps formation and complete inhibition of nematode capturing at 100 mg ai kg⁻¹. Sulphur, triademefon, and tricyclazole showed least toxic effect on conidial trap formation and nematode capturing activities of A. dactyloides in soil up to 100 mg ai kg⁻¹.     

Nanoplastics alter ecosystem multifunctionality and may increase global warming potential

Although the presence of nanoplastics in aquatic and terrestrial ecosystems has received increasing attention, little is known about its potential effect on ecosystem processes and functions. Here, we evaluated if differentially charged polystyrene (PS) nanoplastics (PS-NH₂ and PS-SO₃H) exhibit distinct influences on microbial community structure, nitrogen removal processes (denitrification and anammox), emissions of greenhouse gases (CO₂, CH₄, and N₂O), and ecosystem multifunctionality in soils with and without earthworms through a 42-day microcosm experiment. Our results indicated that nanoplastics significantly altered soil microbial community structure and potential functions, with more pronounced effects for positively charged PS-NH₂ than for negatively charged PS-SO₃H. Ecologically relevant concentration (3 g kg⁻¹) of nanoplastics inhibited both soil denitrification and anammox rates, while environmentally realistic concentration (0.3 g kg⁻¹) of nanoplastics decreased the denitrification rate and enhanced the anammox rate. The soil N₂O flux was always inhibited 6%–51% by both types of nanoplastics, whereas emissions of CO₂ and CH₄ were enhanced by nanoplastics in most cases. Significantly, although N₂O emissions were decreased by nanoplastics, the global warming potential of total greenhouse gases was increased 21%–75% by nanoplastics in soils without earthworms. Moreover, ecosystem multifunctionality was increased 4%–12% by 0.3 g kg⁻¹ of nanoplastics but decreased 4%–11% by 3 g kg⁻¹ of nanoplastics. Our findings provide the only evidence to date that the rapid increase in nanoplastics is altering not only ecosystem structure and processes but also ecosystem multifunctionality, and it may increase the emission of CO₂ and CH₄ and their global warming potential to some extent.     

Effects of heavy metals on aerobic denitrification by strain Pseudomonas stutzeri PCN-1

Effects of heavy metals on aerobic denitrification have been poorly understood compared with their impacts on anaerobic denitrification. This paper presented effects of four heavy metals (Cd(II), Cu(II), Ni(II), and Zn(II)) on aerobic denitrification by a novel aerobic denitrifying strain Pseudomonas stutzeri PCN-1. Results indicated that aerobic denitrifying activity decreased with increasing heavy metal concentrations due to their corresponding inhibition on the denitrifying gene expression characterized by a time lapse between the expression of the nosZ gene and that of the cnorB gene by PCN-1, which led to lower nitrate removal rate (1.67∼6.67 mg L⁻¹ h⁻¹), higher nitrite accumulation (47.3∼99.8 mg L⁻¹), and higher N₂O emission ratios (5∼283 mg L⁻¹/mg L⁻¹). Specially, promotion of the nosZ gene expression by increasing Cu(II) concentrations (0∼0.05 mg L⁻¹) was found, and the absence of Cu resulted in massive N₂O emission due to poor synthesis of N₂O reductase. The inhibition effect for both aerobic denitrifying activity and denitrifying gene expression was as follows from strongest to least: Cd(II) (0.5∼2.5 mg L⁻¹) > Cu(II) (0.5∼5 mg L⁻¹) > Ni(II) (2∼10 mg L⁻¹) > Zn(II) (25∼50 mg L⁻¹). Furthermore, sensitivity of denitrifying gene to heavy metals was similar in order of nosZ > nirS ≈ cnorB > napA. This study is of significance in understanding the potential application of aerobic denitrifying bacteria in practical wastewater treatment.

     

The effects of arsenic and induced-phytoextraction methods on photosynthesis in Pteris species with different arsenic-accumulating abilities

Arsenic (As) accumulation and photosynthesis occur simultaneously in plants under As exposure. We investigated the effects of As and induced-phytoextraction methods on photosynthesis in two As hyperaccumulators (Pteris vittata and Pteris cretica var. nervosa) and two non-hyperaccumulators (Pteris semipinnata and Pteris ensiformis) under soil culture conditions. Chlorophyll fluorescence parameters (the maximum [F_v/F_m] and actual quantum efficiency [F_PSII]) and the activities of three photosynthetic enzymes (ATPase, ribulose-1, 5-bisphosphate carboxylase [RuBPC] and glyceraldehyde-3-phosphate dehydrogenase [GAPDH]) were measured. Arsenic accumulation and photosynthetic behaviours in response to enhanced-phytoextraction methods (trans-1, 2-cyclobexylenedinitrilotetraacetic acid [CDTA] and phosphorous [P] addition and soil pH adjustment) of P. cretica and P. semipinnata were monitored and compared under conditions of 100 mg As kg⁻¹. Significant decreases in the F_v/F_m (19.9%) and F_PSII (36.1%) were observed in P. vittata when exposed to 100 mg As kg⁻¹ in comparison to the control (0 mg As kg⁻¹). Compared to the control (0 mg As kg⁻¹), the activities of GAPDH increased by 0.5% in P. cretica var. nervosa and decreased by only 8.3% in P. vittata even when both of them were treated with 200 mg As kg⁻¹, whereas a significant decrease, 56.1% and 51.7%, of this enzyme was observed in P. semipinnata and P. ensiformis, respectively, when exposed to 50 mg As kg⁻¹. Compared to the control (0 mg CDTA kg⁻¹ or 0 mg P kg⁻¹), the activities of ATPase increased by 53.7% and 82.7% in P. cretica when exposed to 0.5 g CDTA kg⁻¹ and 50 mg P kg⁻¹, respectively, and an increase of up to 175% was also observed in P. semipinnata when exposed to 600 mg P kg⁻¹. The activities of GAPDH increased by 68.9% and 90.7% in P. cretica when exposed to 2 g CDTA kg⁻¹ and 600 mg P kg⁻¹, respectively, but a decrease of up to 60% was observed in P. semipinnata when exposed to 2 g CDTA kg⁻¹. The uptake of As in P. semipinnata increased by 80.9% and 73.3% when 1 g CDTA kg⁻¹ and 600 mg P kg⁻¹ were added, respectively, compared to the control (0 g CDTA kg⁻¹ or 0 mg P kg⁻¹). It was concluded that GAPDH played an important role in the photosynthesis of As hyperaccumulators under As treatments.     

Bioremediation of PAH-contaminated soil with fungi – From laboratory to field scale

The purpose of this study was to develop a fungal bioremediation method that could be used for soils heavily contaminated with persistent organic compounds, such as polyaromatic hydrocarbons (PAHs). Sawmill soil, contaminated with PAHs, was mixed with composted green waste (1:1) and incubated with or without fungal inoculum. The treatments were performed at the laboratory and field scales. In the laboratory scale treatment (starting concentration 3500 mg kg⁻¹, sum of 16 PAH) the high molecular weight PAHs were degraded significantly more in the fungal-inoculated microcosms than in the uninoculated ones. In the microcosms inoculated with Phanerochaete velutina, 96% of 4-ring PAHs and 39% of 5- and 6-ring PAHs were removed in three months. In the uninoculated microcosms, 55% of 4-ring PAHs and only 7% of 5- and 6-ring PAHs were degraded. However, during the field scale (2 t) experiment at lower starting concentration (1400 mg kg⁻¹, sum of 16 PAH) the % degradation was similar in both the P. velutina-inoculated and the uninoculated treatments: 94% of the 16 PAHs were degraded in three months. In the field scale experiment the copy number of gram-positive bacteria PAH-ring hydroxylating dioxygenase genes was found to increase 1000 fold, indicating that bacterial PAH degradation also played an important role.     

Biodegradation of polyacyclic aromatic hydrocarbons in contaminated soil by biostimulation and bioaugmentation in the presence of copper(II) ions

The effect of a combination of biostimulation, bioaugmentation and the presence of CuSO₄ on the biodegradation of the polycyclic aromatic hydrocarbons (PAHs) dibenzofuran 210 mg kg⁻¹, benzo(a)anthracene 200 mg kg⁻¹, dibenzo-p-dioxine 100 mg kg⁻¹ and 9-fluorenone 100 mg kg⁻¹ in a sandy loam soil was studied. The pH of the soil was raised from 5 to 7 with agricultural lime. The soil was then divided into two sets labelled Treatments A and B. Six subsets numbered 1–6 were established for each of Treatments A and B. Treatment A was not amended with CuSO₄ while Treatment B was amended with 5% CuSO₄. Sterile control was established for both sets of treatment by adding 75 g sodium azide twice during the treatment period to Treatment 1. Treatment 2 was established as a natural control, without further amendments. Treatments 3–6 were tilled daily and maintained at 70% moisture. Mono-ammonium phosphate (MAP) (1 g kg⁻¹ soil) was added to Treatments 4–6 twice during the treatment period. H₂O₂ at 500 mg l⁻¹ (600 μl of a 39% solution) was added weekly to Treatment 5. A biosupplement was added to Treatment 6 every week. The experiments were incubated for 10 weeks at ambient temperature. The C:N:P ratio of the soil after the addition of MAP was 38:1:2.2. Counts of microorganisms were larger in the treatments not amended with CuSO₄. For example counts were 77×10⁶ in A6, as compared to 61×10⁶ in B6. CO₂ evolution was also consistently lower in the CuSO₄-amended treatments. Reduction in the concentrations of the PAHs in the treatments without CuSO₄ reached a maximum of 28.6% (dibenzofuran) in the sterile control and 39.1% (dibenzofuran) in the natural control. In contrast, maximum reduction in Treatment 3 reached 85% (dibenzofuran). Reduction in Treatments 4, 5 and 6 was between 90% and 100% in both Treatments A and B after 10 weeks. The combination of biostimulation and bioaugmentation (Treatment 6) enhanced PAH reduction by 100% reduction of all PAHs tested, better than only biostimulation (Treatments 4 & 5), which reduced the PAHs concentrations by between 90% and 100%. The effect of CuSO₄ on the biodegradation of the tested PAHs was more prominent during the early stages of the experiment. However, there was no significant difference in the overall residual concentrations of the PAHs in both treatments A and B at the end of the treatment period.     

Efficiencies of selected biotreatments for the remediation of PAH in diluted bitumen contaminated soil microcosms

Unconventional oils such as diluted bitumen from oil sands differs from most of conventional oils in terms of physiochemical properties and PAHs composition. This raises concerns regarding the effectiveness of current remediation strategies and protocols originally developed for conventional oil. Here we evaluated the efficiency of different biotreatment approaches, such as fungi inoculation (bioaugmentation), sludge addition (bioaugmentation/biostimulation), perennial grasses plantation (phytoremediation) and their combinations as well as natural attenuation (as control condition), for the remediation of soil contaminated by synthetic crude oil (a product of diluted bitumen) in laboratory microcosms. We specifically monitored the PAHs loss percentage (alkylated PAHs and unsubstituted 16 EPA Priority PAHs), the residue of PAHs and evaluated the ecotoxicity of soil after treatment. All treatments were highly efficient with more than?~?80% of ∑PAHs loss after 60 days. Distinctive loss efficiencies between light PAHs (≤?3 rings,?~?96% average loss) and heavy PAHs (4–6 rings,?~?29% average loss) were observed. The lowest average PAHs residue (0.10?±?0.02 mg·kg^?1, for an initial concentration of 0.29?±?0.12 mg·kg^?1) was achieved with the “sludge—plants (grasses)” combination. Sludge addition was the only treatment that achieved significantly lower ecotoxicity (3%?±?4% of growth inhibition of L. sativa) than the control (natural attenuation, 13%?±?4% of inhibition). Sludge addition, grasses plantation and “sludge—fungi combination” treatments could result in lower PAH exposure (than other treatments) in post-treated soil when using the Canadian Soil Quality Guidelines for the protection of environmental and human health for potentially carcinogenic and other PAHs.

     

Role of earthworm-straw interactions on phytoremediation of Cu contaminated soil by ryegrass

It is well known that the earthworm's activities can increase the availability of soil nutrients, improve soil structure, and enhance the biomass of plants in uncontaminated soil. Recently, many researchers found that some metal-tolerant earthworms can survive and even change the fractional distribution of heavy metals in contaminated soil. Furthermore, it has been revealed that earthworms are able to increase metal availability, and therefore, accumulate more metals in plants through their burrowing and casting activity. It is clear that the influence of soil animals is an important factor for phyto-remedation that must be taken into account. ~In this article, the authors studied some effects of addition of earthworms (Metaphire guillelmi), corn straw, and in combinations of earthworms and corn straw on the growth and Cu uptake by ryegrass in Cu contaminated pot soils. The experiment consisted of four levels of Cu addition (0, 100, 200, 400 mg·kg?1) and four treatments. The treatments were 1. control (CK); 2.straw mulching only (M); 3. earthworm additions to soil only (E); and 4.straw mulching plus earthworm additions (ME). Each treatment had three replicates. 10 seeds of ryegrass (Lolium multiflorum) were sowed in each pot and harvested after 30 days. After 30 days of incubation, all earthworms were found to be alive and the pot soils were burrowed through by earthworms. Results showed that the biomass of earthworm declined with the increase of the dosage of Cu additions. The biomass of earthworm increased significantly in treatment 4 (ME) as compared with treatment 3 (E). Not only the earthworms could get more food from the straw, but also could counteract some negative effects of Cu on the earthworms. The rates of straw decomposition in ME treatment increased by about 58.11% ?77.32%. The earthworm activities increased root biomass of ryegrass significantly, but did not show the effect on plant root growth. On the contrary, straw enhanced roots biomass significantly instead of shoots biomass. It was also found that the concentration of Cu in the plant shoot and the plant root, as well as plant Cu uptake were enhanced by earthworm's activities and straw mulching. The increased amount by straw mulching was lower than that of earthworms (E). The treatment of the earthworm–straw mulching combinations enhanced plant Cu concentration, and the amount increased by it was lower than that of the earthworm treatment (E) but higher than that of straw mulching treatment (M). The accumulation factors of copper in the shoots of ryegrass were increased by 31.22% ?121.07%, 2.12% ?61.28% and 25.56% ?132.64%, respectively, in treatment 3(E), 2(M), and 4(ME), respectively. In conclusion, the earthworm activities, straw-mulching and their interactions may have potential roles in elevating phyto-extraction efficiency in low to medium level Cu contaminated soil.     

Characterization of polycyclic aromatic hydrocarbons degradation and arsenate reduction by a versatile Pseudomonas isolate

A Pseudomonas isolate, designated PAHAs-1, was found capable of reducing arsenate and degrading polycyclic aromatic hydrocarbons (PAHs) independently and simultaneously. This isolate completely reduced 1.5 mM arsenate within 48 h and removed approximately 100% and 50% of 60 mg l⁻¹ phenanthrene and 20 mg l⁻¹ pyrene within 60 h, respectively. Using PAHs as the sole carbon source, however, this isolate showed a slow arsenate reduction rate (4.62 μM h⁻¹). The presence of arsenic affected cell growth and concurrent PAHs removal, depending on PAH species and arsenic concentration. Adding sodium lactate to the medium greatly enhanced the arsenate reduction and pyrene metabolism. The presence of the alpha subunit of the aromatic ring-hydroxylating dioxygenase (ARHD) gene, arsenate reductase (arsC) and arsenite transporter (ACR3(2)) genes supported the dual function of the isolate. The finding of latter two genes indicated that PAHAs-1 possibly reduced arsenate via the known detoxification mechanism. Preliminary data from hydroponic experiment showed that PAHAs-1 degraded the majority of phenanthrene (>60%) and enhanced arsenic uptake by Pteris vittata L. (from 246.7 to 1187.4 mg kg⁻¹ As in the fronds). The versatile isolate PAHAs-1 may have potentials in improving the bioremediation of PAHs and arsenic co-contamination using the plant-microbe integrated strategy.     

Bioremediation of Polycyclic Aromatic Hydrocarbons in Creosote-Contaminated Soil by Peniophora incarnata KUC8836

ABSTRACT Polycyclic aromatic hydrocarbons (PAHs) are present in products made from creosote, coal tar, and asphalt. When wood pile treated with creosote is placed in soil, PAHs can contaminate it. Creosote has been used for wood preservation in the past and is composed of approximately 85% PAHs and 15% phenolic compounds. PAHs cause harmful effects to humans and the environment because of their carcinogenic and mutagenic properties. White rot fungi can degrade not only lignin, but also recalcitrant organic compounds such as PAHs. Among numerous white rot fungi used in previous studies, four species were selected to degrade PAHs in a liquid medium. From this evaluation of the degradation of PAHs by the four fungal isolates, two species were ultimately selected for the highest rates of removal. Following 2 weeks of incubation with Peniophora incarnata KUC8836, the degradation rates of phenanthrene, fluoranthene, and pyrene were 86.5%, 77.4%, and 82.6%, respectively. Mycoaciella bispora KUC8201 showed the highest degradation rate for anthracene (61.8%). Hence, bioremediation of creosote-contaminated soil with an initial concentration of 229.49 mg kg^?1 PAHs was carried out using the two selected fungi because they could simultaneously degrade 13 more PAHs than the comparison species. More importantly, isolates of P. incarnata KUC8836 were discovered as powerful degraders of PAHs by producing laccase and manganese-dependent peroxidase (MnP), with 1.7- and 1.1-fold higher than the comparison species, respectively. Therefore, the white rot fungus may be proposed for the removal of PAHs and xenobiotic compounds in contaminated environments.     

Toxicity,tissue accumulation and residue in egg and excreta of copper in laying hens

To study the effects of a high dietary copper dosage on the performance, liver function, and copper residues in the liver, egg and excreta of the laying hen, seventy-five 28-week-old layers were selected and randomly allocated to five dietary treatments with three replications of five birds each for each treatment. The feeding period was 4 weeks followed by a 1 week withdrawal period. The experimental diets were supplemented with 0, 200, 400, 600 or 800 mg kg⁻¹ copper. The response to copper in the diet in terms of feed intake and egg production fitted a quadratic model (P < 0.05), with 200 mg kg⁻¹ giving a positive response and levels of 400 mg kg⁻¹ and above giving a progressively negative response. Enzyme activities, i.e. aspartate aminotransferase (AST), lactate dehydrogenase (LDH), and creatine kinase (CK) in the serum were significantly (P < 0.05) increased with 600 mg of dietary copper inclusion. Histological examination of livers revealed damage by high dosages (over 600 mg) of dietary copper. Copper residues in the liver and excreta were significantly (P < 0.05) increased as dietary copper increased. The copper content of egg reached a maximum at 400 mg kg⁻¹ dietary copper. Serum enzyme activities of AST, LDH and CK returned to normal after the 1 week withdrawal period. Copper residues in the liver, egg and excreta also declined significantly (P < 0.05) during the withdrawal period.     

Phenolic content of Terminalia catappa L. leaf and toxicity evaluation on red hybrid tilapia (Oreochromis sp.)

Despite the use of Terminalia catappa (TC) leaf by traditional fish farmers around the world to improve the health status of cultured fish, there is a paucity of information on comprehensive metabolite profile and the maximum safe dose of the plant. This study aims at profiling the methanol leaf extract of T. catappa, quantifying total phenolic content (TPC) as well as the total flavonoid content (TFC) and evaluating its acute toxicity on blood, plasma biochemical parameters and histopathology of some vital organs in red hybrid tilapia (Oreochromis sp.). The experimental fish were acclimatised for 2 weeks and divided into six groups. Group (1) served as a control group and was administered 0.2 ml,g⁻¹ of phosphate buffer saline (PBS). Groups 2–6 were orally administered T. catappa leaf extracts (0.2 ml.50 g⁻¹) in the following sequence; 31.25, 62.5, 125, 250 and 500 mg.kg⁻¹ body weight. The metabolites identified in T. catappa using liquid chromatography-tandem mass electrospray ionisation spectrometry (LC-ESI-MS/MS) revealed the presence of organic acids, hydrolysable tannins, phenolic acids and flavonoids. Phenolic quantification revealed reasonable quantity of phenolic compounds (217.48 μg GAEmg⁻¹ for TPC and 91.90 μg. QCEmg⁻¹ for TFC). Furthermore, there was no significant difference in all the tested doses in terms of blood parameters and plasma biochemical analysis except for the packed cell volume (PCV) at 500 mg.kg⁻¹ when compared to the control. Significant histopathological changes were observed in groups administered with the extract at 125, 250 and 500 mg.kg⁻¹ doses. To a very large extent it is therefore safe to administer the extract at 31.25 and 62.5 mg.kg⁻¹ in tilapia.     

Long-term changes of ecosystem services at Solling,Germany: Recovery from acidification,but increasing nitrogen saturation?

Time series of values of ingenious parameters indicating ecosystem services from European beech and Norway spruce ecosystems at Solling, Germany, were evaluated with respect to resilient or adaptive behaviour. Studied indicators comprise the use of monitoring data with up to more than 40 years of observation on deposition of potential acidity, sulphate (SO₄²⁻) budgets, exchangeable base cation pools, Bc/Al ratio in soil solution, nitrogen (N) budgets, foliar nutrition as indicated by the foliar Bc/N ratio, and defoliation. Deposition of potential acidity decreased considerably at both ecosystems. SO₄²⁻ budgets reveal retention of sulphur in the soils affecting acid/base budgets. Exchangeable base cation pools decreased at both ecosystems by about 60%. Bc/Al ratio in soil solution in the mineral soil was mostly below critical limits indicating potential toxic stress to tree roots. N retention in the soils decreased from about 40 kg ha⁻¹ yr⁻¹ in the 1970s to currently very low rates of 0–20 kg ha⁻¹ yr⁻¹ indicating increasing N saturation. Foliar Bc/N ratio decreased at the spruce ecosystem indicating possible nutrient imbalances. Defoliation at both Solling ecosystems is on a high level compared to other forests in Germany, but reveals no distinct relation to soil acidification or N saturation. From the selected indicators, SO₄²⁻ and N budgets reveal resilient behaviour, whereas indicators related to the acid/base status tend to adaptive behaviour.     

Influence of the fungicide pentachloronitrobenzene on VA-mycorrhizal and total root length and phosphorus uptake of oats (Avena sativa)

The effect of application of the fungicide pentachloronitrobenzene (PCNB) at levels between 2 and 50 mg kg^–1 soil on root growth, mycorrhizal infection and P uptake was studied in pot culture with oats (Avena sativa cv. Alfred) growing in a rendzina soil low in available P. The soil had been partially sterilized by X-ray, and half of the pots were inoculated with spores of the VAM-fungusGlomus mosseae (indigenous species).Soil irradiation (0.5 Mrad) did not decrease the levels of infection by VAM. Application of PCNB decreased the VAM-infected root length, at 50 mg PCNB kg^–1 soil VAM-infected root length was about 12% of the controls. Total root length, however, increased to about 126% of control values at PCNB rates up to 20 mg kg^–1 soil, but decreased to 89% of the controls at 50 mg kg^–1 soil. Total P-uptake decreased with increasing levels of PCNB and was linearly correlated with infected root length (r=0.92).The stimulation of root growth by PCNB at rates up to 20 mg kg^–1 soil is regarded as an indirect effect, brought about by suboptimal P-supply due to inhibition of VA-mycorrhiza. Conversely, the reduction of total root length at 50 mg PCNB kg^–1 soil is most likely a direct effect. Due to the phytotoxicity of the fungicide, the contribution of the indigenous VA-mycorrhiza to plant P uptake under field conditions cannot be determined by soil application of PCNB at rates sufficient for complete inhibition of VAM.As inhibition or absence of VAM may lead to compensatory root growth, mycorrhizal dependency ought to be calculated from the amounts of P taken up per unit root length in mycorrhizal and nonmycorrhizal plants, respectively.     

The effects of copper, iron and zinc on digestive enzyme activity in the hybrid tilapia Oreochromis niloticus (L.) ×Oreochromis aureus (Steindachner)

The present experiment was conducted to study effects of Cu, Fe and Zn on activities of digestive enzymes of the hybrid tilapia Oreochromis niloticus×Oreochromis aureus. The acidic protease activities increased 65·5 and 55·1% by addition of homogenates of digesta‐containing stomach with copper (75 mg l⁻¹) and zinc (50 mg l⁻¹) respectively. Addition of Cu and Zn increased the activities of protease in the hepatopancreas homogenates by 132·7 and 38·1% respectively, and reduced the activity of protease in the digesta‐containing intestine homogenates by 11·0 and 13·8% respectively. Addition of Fe (50 mg l⁻¹) increased the acidic protease activity by 96·7% but did not alter the activities of protease in the intestine and hepatopancreas. Addition of Cu markedly inhibited activities of amylase in intestine and hepatopancreas homogenates, while Zn addition showed no effects. Addition of Fe reduced activities of amylase in the intestine homogenates by 47·9% but had no effect on amylase activities in the hepatopancreas. When Cu (75 mg kg⁻¹), Fe (50 mg kg⁻¹) and Zn (50 mg kg⁻¹) were supplemented to basal diet for 3 weeks, the activities of amylase in hepatopancreas homogenates increased 125·3, 215·6 and 70·0%, respectively, the activities of amylase in intestine increased 79·8, 74·6 and 48·5%, respectively, and the activities of lipase in intestine increased 90·5, 149·8 and 84·0%, respectively. Supplementation of Cu, Fe or Zn into diet had no effects on activity of protease in all digestive organs. Therefore, the results suggest that effects of Cu, Fe and Zn on activity of digestive enzymes in vitro were different from those seen in vivo, and that the positive effects of Cu, Fe and Zn supplemented to fish diet would be valuable information for formulating fish feed.