Evaluation of phenanthrene toxicity on earthworm (Eisenia fetida): An ecotoxicoproteomics approach

The goal of this study was to identify promising new biomarkers of phenanthrene by identifying differentially expressed proteins in Eisenia fetida after exposure to phenanthrene. Extracts of earthworm epithelium collected at days 2, 7, 14, and 28 after phenanthrene exposure were analyzed by two dimensional electrophoresis (2-DE) and quantitative image analysis. Comparing the intensity of protein spots, 36 upregulated proteins and 45 downregulated proteins were found. Some of the downregulated and upregulated proteins were verified by MALDI-TOF/TOF-MS and database searching. Downregulated proteins in response to phenanthrene exposure were involved in glycolysis, energy metabolism, chaperones, proteolysis, protein folding and electron transport. In contrast, oxidation reduction, oxygen transport, defense systems response to pollutant, protein biosynthesis and fatty acid biosynthesis were upregulated in phenanthrene-treated E. fetida. In addition, ATP synthase b subunit, lysenin-related protein 2, lombricine kinase, glyceraldehyde 3-phosphate dehydrogenase, actinbinding protein, and extracellular globin-4 seem to be potential biomarkers since these biomarker were able to low levels (2.5 mg kg⁻¹) of phenanthrene. Our study provides a functional profile of the phenanthrene-responsive proteins in earthworms. The variable levels and trends in these spots could play a potential role as novel biomarkers for monitoring the levels of phenanthrene contamination in soil ecosystems.     

The effects of ethylene chlorohydrin on fatty acid synthesis 1 2 3

Abstract

Male chicks weighing 700 to 900 g. received an acute or eight doses IG of 60 or 40 mg/kg ethylene chlorohydrin (ECH) respectively and were sacrificed eighteen hours after the last dose. Mitochondrial elongation of fatty acids was decreased significantly while fatty acid synthetase activity was not significantly affected by ECH treatment. Cytochrome c oxidase activity in fresh whole liver homogenate was significantly higher in chicks subjected to acute exposure with ECH when compared to the controls. Upon freezing and thawing of homogenates, cytochrome c oxidase activity increased significantly in the control group but was unchanged in the ECH group which suggests that the mitochondrial membrane integrity is compromised by the ECH treatment. Serum and liver triglyceride levels were significantly elevated in both the single and multiple ECH dose groups. Liver to body weight ratios were significantly higher in both treatment groups when compared to their controls. Histological examination of the liver of ECH‐treated chicks showed cytoplasmic clearing of the cells but no vacuolization or centri‐lobular necrosis. Serum isocitrate dehydrogenase levels were significantly higher in the multiple treatment ECH group than in the control group.     

Growth and physiological responses of Pennisetum sp. to cadmium stress under three different soils

Pennisetum sp. was employed as a model species to detect the growth and physiological response to cadmium (Cd) stress at different Cd concentrations (0, 20, 50, and 100 mg kg⁻¹) in three types of soils (yellow brown soil, yellow soil, and red soil). Results showed that the growth of Pennisetum sp. was not significantly influenced by Cd in 20 mg kg⁻¹, but significantly inhibited at higher Cd concentrations in three types of soils. Besides, the higher Cd concentrations, the lower root, stem, and leaf biomass. With Cd concentration of soil increasing, Cd content of root, stem, and leaf increased. Compared with no Cd, high Cd concentrations (50 and 100 mg kg⁻¹) induced the physiological indices (photosynthetic rate, stomatal conductance, transpiration rate) and biochemical indices (nitrate reductase, glutamine synthetase, and glutamate synthase activities) decreasing, but the concentration of NO₃⁻ and NH₄⁺ increasing. The activity of antioxidative enzymes (SOD, POD, and CAT) was disrupted and the content of malondialdehyde (MDA) increasing. Pennisetum sp. could protect cells from damage and maintain normal physiological metabolism via increasing the production of soluble sugar and soluble protein, but soluble proteins and soluble sugars were limited in high concentrations of Cd (50 and 100 mg kg⁻¹). Moreover, the growth and physiological response to Cd are different in the three types of soils. The growth of Pennisetum sp. in yellow brown soil was better than that in other two soils, and the gas exchange rate, antioxidant enzyme activity, and nitrogen metabolism in yellow soil and red soil were more affected by Cd stress than that in yellow brown soil. Overall, Pennisetum sp. had certain tolerance and biosorption ability to Cd in different Cd concentrations and different types of soil. Hence, Pennisetum sp. was a suitable choice for Cd remediation, especially in yellow brown soil.

     

Differential expression profile of membrane proteins in Aplysia pleural–pedal ganglia under the stress of methyl parathion

This study was aimed to analyze the alteration of membrane protein profiles in Aplysia juliana Quoy & Gaimard (A. juliana) pleural–pedal ganglia under MP exposure. Both the results of GC–MS analysis and the activity assay of acetylcholinesterase (AChE), superoxide dismutase (SOD), catalase (CAT) reveal that MP toxicological effects on Aplysia left and right pleural–pedal ganglia are different under 7 and 14 days of exposure. Therefore, Aplysia were subjected for exposure at two concentrations (1 and 2 mg/l) of MP for 7 and 14 days for membrane proteomic study. As a result, 19 and 14 protein spots were differentially expressed in A. juliana left pleural–pedal ganglia under 7 and 14 days treatment, and 20 and 14 protein spots found with differential expressions in their right ganglia under the same treatment, respectively. Several proteins with expression variations were detected from both the left and right pleural–pedal ganglia; however, most proteins have distinctive expressions, indicating different mechanisms might be involved in initiating MP toxicology in left and right ganglia. Among the total differential protein spots obtained, 29 proteins were classed as membrane proteins. These proteins are mainly involved in the metabolism process, cell redox homeostasis, signal transduction, immunology, intracellular transport and catalysis, indicating MP toxicity in mollusks seems to be complex and diverse. Some differentially expressed proteins were further confirmed by Western blotting and quantitative real-time PCR. These results might provide renovated insights to reveal the mechanism of MP-induced neurotoxicity, and the novel candidate biomarkers might have potential application for environmental evaluation of MP pollution level.     

Investigation of arsenic accumulation and tolerance potential of Sesuvium portulacastrum (L.) L

Sesuvium portulacastrum (L.) L., a facultative halophyte, is considered a suitable candidate for the phytoremediation of metals. An investigation of As accumulation and tolerance was conducted in Sesuvium plants upon exposure to As(V) (100-1000 μM) for 30 d. Plants demonstrated a good growth even after prolonged exposure (30 d) to high As(V) concentrations (1000 μM) and a significant As accumulation (155 μg g⁻¹ dry weight) with a bioaccumulation factor of more than ten at each concentration. The results of shoot and root dry weight, malondialdehyde accumulation, photosynthetic pigments, and total soluble proteins demonstrated that plants did not experience significant toxicity even at 1000 μM As(V) after 30 d. However, metabolites (total non-protein thiols and cysteine) and enzymes (serine acetyltransferase, cysteine synthase and γ-glutamylcysteine synthetase) of thiol metabolism, in general, remained either unaffected or showed slight decline. Hence, plants tolerated high As(V) concentrations without an involvement of thiol metabolism as a major component. Taken together, the results indicate that plants are potential As accumulator and may find application in the re-vegetation of As contaminated sites.     

Antimicrobial effects of clofibrate on the wheat pathogen Fusarium culmorum

Abstract

The biological effects of clofibrate (ethyl p‐chlorophenoxy‐isobutyric acid) on the growth and metabolism of the soil‐borne wheat pathogen Fusarium culmorum, were examined.

In mid log phase (16 hr) cultures both phenylalanine uptake and secondary spore production were stimulated at 0.1 μM concentration; the net sterol content was reduced 50% at 0.35 μM; oxygen uptake was stimulated at 0.1 mM; growth was inhibited 50% at 0.1 mM concentration. Both phenylalanine and oxygen uptake were inhibited at 1.0 mM and pyruvate dehydrogenase activity was reduced 50% at 50 mM concentration of clofibrate.

The data indicate that clofibrate affects a number of biological and enzyme systems. The inhibitory effect on the growth of the pathogen suggest a potential use of hypolipidemic agents like clofibrate as an antifungal agent for seed protection.     

Proteomic changes in <Emphasis Type="Italic">Corbicula fluminea</Emphasis> exposed to wastewater from a psychiatric hospital

The increase use of pharmaceutical compounds in veterinary practice and human population results in the ubiquitous presence of these compounds in aquatic ecosystems. Because pharmaceuticals are highly bioactive, there is concern about their toxicological effects in aquatic organisms. Therefore, the aim of this study was to assess the effects of an effluent from a psychiatric hospital (containing a complex mixture of 25 pharmaceutical compounds from eleven therapeutic classes) on the freshwater clam Corbicula fluminea using a proteomic approach. The exposure of C. fluminea to this complex effluent containing anxiolytics, analgesics, lipid regulators, beta blockers, antidepressants, antiepileptics, antihistamines, antihypertensives, antiplatelets and antiarrhythmics induced protein changes after 1 day of exposure in clam gills and digestive gland more evident in the digestive gland. These changes included increase in the abundance of proteins associated with structural (actin and tubulin), cellular functions (calreticulin, proliferating cell nuclear antigen (PCNA), T complex protein 1 (TCP1)) and metabolism (aldehyde dehydrogenase (ALDH), alcohol dehydrogenase, 6 phosphogluconate dehydrogenase). Results from this study indicate that calreticulin, PCNA, ALDH and alcohol dehydrogenase in the digestive gland and T complex protein 1 (TCP1)) and 6 phosphogluconate dehydrogenase in the gills represent useful biomarkers for the ecotoxicological characterization of psychiatric hospital effluents in this species.     

Emission characteristics for co-combustion of leather wastes,sewage sludge,and coal in a laboratory-scale entrained flow tube furnace

Four different mixed fuels consisted of leather waste, coal, and sewage sludge were combusted in a lab-scale entrained flow fluidized bed furnace. The influence of blending ratio on emission characteristics of SO₂, NO_x, HCl, particulate matter (PM), heavy metals, and polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/Fs) was studied. Results showed that the mixing of coal with sewage sludge had a complex effect on the emission characteristics. On the one hand, with more sewage sludge blending in the mixed fuel, the acid gas pollutant (SO₂, NO_x) decreased a lot, and the recovery of volatile heavy metals (Cd, Pb) increased at the same time. Furthermore, the leaching toxicity of Cr in the fly ash and bottom ash went down below the national standard with the adding of sewage sludge. On the other hand, the mixing of sewage sludge which consisted of more ash content resulted in the increase of the PM emission. Moreover, the high content of Cu and chlorine in the sewage sludge can promote the formation of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/Fs) when the fuel 3 and 4 were combusted. Most importantly, the concentration of toxic PCDD/Fs in the flue gas produced from fuel 3 and fuel 4 was successfully controlled down below 0.20 ng I-TEQ/Nm³ by the active carbon.

     

Alterations in liver marker enzymes and recovery ability of cyprinids as indicators of aquatic pollution

Information on the detection of pesticides in fresh water Lakes of Kashmir (Dal and Mansbal) through GC–MS technique is scarce, and also the recovery in biochemical parameters (AST, ALT and ALP) of fish after transferring them to clean media has not been reported yet. The water samples were collected from three sites and analyzed for their pesticide profile by dispersive liquid–liquid micro extraction (DLLME) followed by GC–MS. Influence of pesticides on liver marker enzymes of Cyprinus carpio and Carassius carassius was also investigated. The results obtained showed the presence of three main pesticides viz. chlorpyrifros, dimethoate and dichlorvos in waters of Dal Lake whereas no pesticide was detected in waters of Mansbal. The higher values for AST, ALT and ALP activities and decrease in protein content were obtained in the samples from the Dal Lake compared with those from Mansbal Lake (p < 0.05). These data when compared with the values found in C. carpio and C. carassius from both the Dal Lake and Mansbal Lake kept under laboratory conditions after 15, 30, 45 and 60 days of maintenance in clean media found that during depuration, all the enzyme activities came down significantly (p < 0.05) indicating the compensatory response by the fish against the pesticide stress. Therefore, these parameters could be used as indicators of pesticide pollution in aquatic organisms and were recommended for environmental monitoring for investigating the mechanism involved in the recovery pattern.

     

MSMA Resistance Studies

Abstract

Monosodium methanearsonate (MSMA)-resistant and -susceptible common cocklebur (Xanthium strumarium L.) and cotton (Gossypium hirsutum L.) were treated with MSMA. Plant parameters analyzed were: glutathione synthetase activity, selected amino acid (arginine, glutamic acid, alanine, citrulline, glutamine, and glutathione) content and arsenic content (MSMA, total arsenic, and arsonate). No reduction of arsenic from the parent pentavalent form present in MSMA to the trivalent form was detected. Arginine, glutamic acid, and glutamine content increased in tissue three days after MSMA treatment. Glutathione content decreased during the first three days after treatment; however, five days after treatment the resistant biotype of cocklebur and cotton had elevated glutathione levels (8–20 times greater, respectively). Glutathione Synthetase activity was higher in cotton than in either of the cocklebur biotypes; MSMA did not affect its activity in cotton or either cocklebur biotype. Resistant biotypes have a slightly higher activity than the susceptible biotype. Tolerance of cotton to MSMA may be related to glutathione synthetase activity and possibly to the presence of phytochelatins. Also, increased glutathione levels in the resistant biotype may implicate phytochelatin involvement in the resistance mechanism.     

Enhancing mixed toluene and formaldehyde pollutant removal by Zamioculcas zamiifolia combined with Sansevieria trifasciata and its CO2 emission

Indoor air pollutants comprise both polar and non-polar volatile organic compounds (VOCs). Indoor potted plants are well known for their innate ability to improve indoor air quality (IAQ) by detoxification of indoor air pollutants. In this study, a combination of two different plant species comprising a C3 plant (Zamioculcas zamiifolia) and a crassulacean acid metabolism (CAM) plant (Sansevieria trifasciata) was used to remove polar and non-polar VOCs and minimize CO₂ emission from the chamber. Z. zamiifolia and S. trifasciata, when combined, were able to remove more than 95% of pollutants within 48 h and could do so for six consecutive pollutant’s exposure cycles. The CO₂ concentration was reduced from 410 down to 160 ppm inside the chamber. Our results showed that using plant growth medium rather than soil had a positive effect on decreasing CO₂. We also re-affirmed the role of formaldehyde dehydrogenase in the detoxification and metabolism of formaldehyde and that exposure of plants to pollutants enhances the activity of this enzyme in the shoots of both Z. zamiifolia and S. trifasciata. Overall, a mixed plant of Z. zamiifolia and S. trifasciata was more efficient at removing mixed pollutants and reducing CO₂ than individual plants.

     

Co-digestion of sewage sludge and dewatered residues from enzymatic hydrolysis of sugar beet pulp

Sugar beet pulp residues (SBPR) from hydrolysis and dewatering of beet pulp were co-digested with municipal sewage sludge (MSS). The highest biogas yields of nearly 512 dm³/kg VS_fed (volatile solids fed) were achieved for SBPR, treated both as the monosubstrate and as a mixture with MSS (1 : 1 by weight). Simultaneously, the highest methane production of 348 dm³ CH₄/kg VS_fed was determined when the sewage sludge was co-digested with 35% SBPR. The analysis of digestate showed that neither ammonia nor volatile fatty acids destabilized the biogas production.

Implications:?Processing of sugar beet pulp into bioethanol via enzymatic hydrolysis and microbial fermentation has become increasingly attractive. However, in this process, only the liquid fraction derived from hydrolysis is subjected into alcoholic fermentation, whereas the remaining solid fraction needs to be utilized. This study demonstrated that sugar beet pulp residues after bioethanol production can successfully be co-digested with sewage sludge to increase biogas productivity of anaerobic digesters located at wastewater treatment plants.     

Proteomic analysis of chromate-induced modifications in Pseudokirchneriella subcapitata

In this work, we have analyzed the changes in the protein expression profile elicited by chromium (Cr) exposure in the freshwater green alga Pseudokirchneriella subcapitata, a well known bio-indicator of water pollution. We tested two experimental conditions, namely 0.2 and 1 ppm of potassium dichromate; this concentration range includes the environmentally-relevant concentrations. Results show that neither concentration of potassium dichromate tested inhibited algal growth. However, the proteomic approach allowed the identification of relevant modifications in protein expression. In fact, among 800 protein spots detected by two-dimensional electrophoresis, 16 Cr-regulated proteins, including predicted and novel ones, were identified using tandem mass spectromic protein analysis.The results demonstrate a Cr-specific action in altering several photosynthetic proteins, such as ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO), RuBisCO activase, Light Harvesting Chla/b protein complex, and stress related Chla/b binding protein1. Although Cr toxicity with respect to photosynthesis has been already documented, here we have identified, for the first time, the target proteins of this toxicity. Cr also induced a modulation of some proteins involved in the metabolism of the amino acids glutamine, arginine and methionine. These data are supported by changes in cellular polyamine (PA) accumulation. Present findings provide new insight into the molecular mechanisms underlying Cr toxicity in P. subcapitata.     

氧化亚铁硫杆菌对电动力处理城市污泥中重金属影响研究

分析了氧化亚铁硫杆菌对污泥中重金属形态及对电动力处理污泥中重金属的去除率影响.氧化亚铁硫杆菌处理后的污泥,pH值降低,部分重金属从稳定态转化为可溶态.经氧化亚铁硫杆菌预处理后的泥样和未经预处理的泥样,在同样条件下用电动力处理,处理结果为:预处理后Cd的去除率从61.85%上升到72.54%,Cu从34.26%上升到63.51%,Zn从65.06%上升到7L 22%.预处理对Cu的影响比Cd、Zn大得多.用电动力处理Cu超标污泥中重金属时,将污泥先用氧化亚铁硫杆菌预处理,可有效地提高其去除效果,处理后的污泥有利于农用.     

The metabolic fate of 14C‐parathion by some fresh water phytoplankton and its possible effects on the algal metabolism

Abstract

The growth and total carbohydrate contents of Nostoc muscorum and Tolypothrix tenuis were greatly and significantly reduced by the application of parathion. “Chlorophyll a”, carotene biosynthesis and the rate of glucose absorption were enhanced after supplementation of parathion to the culture media of both cyanobacteria. Nitrogen released to the media, total nitrogen content and total nitrogen fixed were increased in both organisms‐ Increase in protein content was accompanied by remarkable drop in amino, peptide and ammonia fractions‐ Phosphorus uptake, RNA, DNA and total phosphorus content were accelerated to reach maximum accumulation at the highest insecticide level. In metabolism study using ¹⁴C‐labelled compound, parathion was readily degraded by Nostoc and Tolypothrix. Following ten days incubation, the aqueous fractions contained 21.1% and 18.1% of the initial activity in Nostoc and Tolypothrix respectively. TLC analysis of the hydrolytic products revealed the presence of three metabolites: p‐aminophenol, p‐nitrophenol and aminoparathion.     

The effects of different sewage sludge amendment rates on the heavy metal bioaccumulation,growth and biomass of cucumbers (Cucumis sativus L.)

When sewage sludge is incorrectly applied, it may adversely impact agro-system productivity. Thus, this study addresses the reaction of Cucumis sativus L. (cucumber) to different amendment rates (0, 10, 20, 30, 40 and 50 g kg^?1) of sewage sludge in a greenhouse pot experiment, in which the plant growth, heavy metal uptake and biomass were evaluated. A randomized complete block design with six treatments and six replications was used as the experimental design. The soil electrical conductivity, organic matter and Cr, Fe, Zn and Ni concentrations increased, but the soil pH decreased in response to the sewage sludge applications. As approved by the Council of European Communities, all of the heavy metal concentrations in the sewage sludge were less than the permitted limit for applying sewage sludge to land. Generally, applications of sewage sludge of up to 40 g kg^?1 resulted in a considerable increase in all of the morphometric parameters and biomass of cucumbers in contrast to plants grown on the control soil. Nevertheless, the cucumber shoot height; root length; number of leaves, internodes and fruits; leaf area; absolute growth rate and biomass decreased in response to 50 g kg^?1 of sewage sludge. All of the heavy metal concentrations (except the Cu, Zn and Ni in the roots, Mn in the fruits and Pb in the stems) in different cucumber tissues increased with increasing sewage sludge application rates. However, all of the heavy metal concentrations (except the Cr and Fe in the roots, Fe in the leaves and Cu in the fruits) were within the normal range and did not reach phytotoxic levels. A characteristic of these cucumbers was that all of the heavy metals had a bioaccumulation factor <1.0. All of the heavy metals (except Cd, Cu and Zn) had translocation factors that were <1.0. As a result, the sewage sludge used in this study could be considered for use as a fertilizer in cucumber production systems in Saudi Arabia and can also serve as a substitute method of sewage sludge disposal.

The effects of different sewage sludge amendment rates on the heavy metal bioaccumulation, growth and biomass of cucumbers

     

Alterations of protein profile in zebrafish liver cells exposed to methyl parathion: a membrane proteomics approach

Methyl parathion (MP) is an extensively used organophosphorus pesticide, which has been associated with a wide spectrum of toxic effects on environmental organisms. The aim of this study is to investigate the alterations of membrane protein profiles in zebrafish liver (ZFL) cell line exposed to MP for 24 h using proteomic approaches. Two-dimensional gel electrophoresis revealed a total of 13 protein spots, whose expression levels were significantly altered by MP. These differential proteins were subjected to matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry analysis, and nine proteins were identified to be membrane proteins, among which seven were up-regulated, while two were down-regulated. In addition, the mRNA levels corresponding to these differential membrane proteins were further analyzed by quantitative real-time PCR. And the differential expression of arginase-2 was specially validated via Western blotting. Regarding the physiological functions, these proteins are involved in molecular chaperon, cytoskeleton system, cell metabolism, signal transduction, transport and hormone receptor respectively, suggesting the complexity of MP-mediated toxicity to ZFL cell. These data could provide useful insights for better understanding the hepatotoxic mechanisms of MP and develop novel protein biomarkers for effectively monitoring MP contamination level in aquatic environment.     

Biochemical profiling of lead-intoxicated impaired lipid metabolism and its amelioration using plant-based bioactive compound

The aim of this study was to investigate the lead (Pb)-induced lipid metabolism impairment and its amelioration using plant-based therapeutic interventions. Pb-induced hepatotoxicity can disturb the normal levels of natural antioxidant enzymes including glutathione (GSH) and superoxide dismutase (SOD) exerting a crucial impact on membrane unsaturated fatty acids (FA), hence leading to lipid peroxidation. Furthermore, Pb toxicity can also alter the regulation of various hormones involved in the synthesis of 3-hydroxy-methyl glutaryl CoA (HMG-CoA reductase), leading to an impairment in normal levels of serum cholesterol and other associated conjugated lipid molecules such HDL-cholesterol, LDL-cholesterol and VLDL-cholesterol. In this study, the lipoprotein fractions, cholesterol, triglyceride (TGs) and biomarkers of liver functions were estimated by employing respective assay kits. The levels of antioxidant enzymes, FFAs and HMG-CoA reductase were determined by employing sandwich ELISA method. The administration of PbAc in experimental rats induced a significant disturbance in lipid profile (P < 0.05) accompanying a significant reduction in natural antioxidant defence system (P < 0.05). The significant alteration in the levels of serum antioxidant enzymes can lead to membrane lipid peroxidation that is reflected by a significantly (P < 0.05) high level of serum MDA in PbAc-induced experimental rats. However, the administration of resveratrol proved therapeutically effective in the treatment of Pb toxicity. Overall, the results of this study accompanying histopathological examination had proved the ameliorating effect of resveratrol in Pb-induced lipid metabolism impairment by adopting vitamin C as a standard therapeutic intervention.