Oxidative stress and hepatotoxicity in the frog,Rana chensinensis,when exposed to low doses of trichlorfon

Trichlorfon is an organophosphate insecticide that is widely used in aquaculture and agriculture against parasitic infestations and has caused aquatic toxicity to non-target organisms. To evaluate the effects of low doses of trichlorfon on the oxidative stress and hepatotoxicity in amphibians, Chinese brown frogs (Rana chensinensis) were exposed to trichlorfon at concentrations of 0, 0.01, 0.1, and 1.0 mg/L for 2 and 4 weeks. Then, the activities of superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST), and the content of malondialdehyde (MDA) in hepatic tissue were examined to evaluate the effects of oxidative stress and lipid peroxidation. The histopathological alternations to the liver were observed through light and transmission electron microscopy (TEM). The results showed that SOD and CAT activities were increased in the livers of frogs exposed to various concentrations of trichlorfon. The GST activity showed no significant changes at any concentration after 2 weeks of exposure, whereas there was an initial increase after exposure to 0.1 mg/L of trichlorfon at 4 weeks. The content of MDA revealed a significant decrease after exposure. Histopathological and ultrastructural studies showed that trichlorfon induced hyalinization, vacuolation, nucleus necrosis, and cellular swelling in hepatocytes. These results suggest that low doses of trichlorfon could induce oxidative stress, lipid peroxidation, and hepatic lesions in frogs, which shows that even lower, non-lethal doses of trichlorfon are potentially toxic to amphibians.     

Formetanate toxicity and changes in antioxidant enzyme system of <Emphasis Type="Italic">Apis mellifera</Emphasis> larvae

Substantial percentage of world food production depends on pollinating service of honeybees that directly depends on their health status. Among other factors, the success of bee colonies depends on health of developed larvae. The crucial phase of larval development is the first 6 days after hatching when a worker larva grows exponentially and larvae are potentially exposed to xenobiotics via diet. In the present study, we determined the lethal concentration LC₅₀ (72 h) following single dietary exposure of honeybee larvae to formetanate under laboratory conditions, being also the first report available in scientific literature. Activities of antioxidant enzymes, superoxide dismutase (SOD), catalase (CAT) and glutathione-S-transferase (GST) were also measured in the homogenates of in vitro reared honeybee larvae after single formetanate exposure. Decreased specific activity of SOD and increased activities of CAT and GST suggest the induction of oxidative stress. Higher levels of thiobarbituric reactive species in all samples supported this fact. Comparing determined larval toxicity (LC₅₀ of 206.01 mg a.i./kg diet) with adult toxicity data, we can suppose that the larvae may be less sensitive to formetanate than the adult bees.     

Effects of malathion and chlorpyrifos on acetylcholinesterase and antioxidant defense system in Oxya chinensis (Thunberg) (Orthoptera: Acrididae)

The study was undertaken to evaluate the effects of malathion and chlorpyrifos on acetylcholinesterase (AChE), esterase (EST) activity and antioxidant system after topical application with different concentration to Oxya chinensis. The results showed that malathion and chlorpyrifos inhibited EST, AChE activity and increased malondialdehyde (MDA) contents. A change in superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST), glutathione peroxidase (GPx), and glutathione reductase (GR) activity combined with reduced glutathione (GSH) and total glutathione (tGSH) contents was found in O. chinensis after malathion and chlorpyrifos treatments. Malathion and chlorpyrifos increased SOD and CAT activity compared with the control. With the concentrations increasing, SOD and CAT activity showed the similar tendency, namely, SOD and CAT activity increased at the lower concentrations and decreased at the higher concentrations. The results showed that malathion and chlorpyrifos decreased significantly GR activity. GST and GPx activity at the studied concentrations of chlorpyrifos was lower than that of the control. However, no significance was observed. GPx and GST activity in malathion treated grasshoppers showed a biphasic response with an initial increase followed by a decline in its activity. Malathion and chlorpyrifos decreased GSH contents and the ratio of GSH/GSSG. The present findings indicated that the toxicity of malathion and chlorpyrifos might be associated with oxidative stress.     

Imidacloprid induced alterations in oxidative stress,biochemical, genotoxic,and immunotoxic biomarkers in non-mammalian model organism Galleria mellonella L. (Lepidoptera: Pyralidae)

Imidacloprid (IMI), a neonicotinoid insecticide, is widely used to control pests in agriculture. We investigated the changes in antioxidant enzyme activities, lipid peroxidation levels, biochemical effects, genotoxic effect, and immunotoxic effect of sublethal doses (0.25, 0.50, 0.75, and 1.00?µg) of IMI at different time periods (24, 48, 72, and 96?h) on a model organism, Galleria mellonella L. The results indicated that there were dose-dependent increases in both antioxidant enzyme activities (SOD and CAT) and MDA levels. Protein content was not affected by IMI at 24th and 48th, whereas it was decreased by the highest dose of IMI (1.00?µg) at 72nd and 96th h. Lipid and carbohydrate contents were reduced with increasing doses of IMI. Micronucleus frequency significantly increased in all IMI doses. All IMI doses caused a significant decrease in THC at 24th, 48th, and 72nd h. Our results can help to illustrate the effects of IMI in target organisms and indirectly may aid to discover potential risk of it on nontarget organisms. Future studies, at molecular levels, will be helpful in understanding the mechanism of action of IMI on these biomarkers.     

Responses of earthworm to aluminum toxicity in latosol

Excess aluminum (Al) in soils due to acid rain leaching is toxic to water resources and harmful to soil organisms and plants. This study investigated adverse impacts of Al levels upon earthworms (Eisenia fetida) from the latosol (acidic red soil). Laboratory experiments were performed to examine the survival and avoidance of earthworms from high Al concentrations and investigate the response of earthworms upon Al toxicity at seven different Al concentrations that ranged from 0 to 300 mg kg^?1 over a 28-day period. Our study showed that the rate of the earthworm survival was 100 % within the first 7 days and decreased as time elapsed, especially for the Al concentrations at 200 and 300 mg kg^?1. A very good linear correlation existed between the earthworm avoidance and the soil Al concentration. There was no Al toxicity to earthworms with the Al concentration ≤50 mg kg^?1, and the toxicity started with the Al concentration ≥100 mg kg^?1. Low Al concentration (i.e., <50 mg kg^?1) enhanced the growth of the earthworms, while high Al concentration (>100 mg kg^?1) retarded the growth of the earthworms. The weight of earthworms and the uptake of Al by earthworms increased with the Al concentrations from 0 to 50 mg kg^?1 and decreased with the Al concentrations from 50 to 300 mg kg^?1. The protein content in the earthworms decreased with the Al concentrations from 0 to 100 mg kg^?1 and increased from 100 to 300 mg kg^?1. In contrast, the catalase (CAT) and superoxide dismutase (SOD) activities in the earthworms increased with the Al concentrations from 0 to 100 mg kg^?1 and decreased from 100 to 300 mg kg^?1. The highest CAT and SOD activities and lowest protein content were found at the Al concentration of 100 mg kg^?1. Results suggest that a high level of Al content in latosol was harmful to earthworms.     

Effect of heavy metals on tissue-specific antioxidant response in Indian major carps

Concentrations of heavy metals (Cu, Ni, Zn, Cd and Pb) were measured in sediments, water and liver and kidney tissues of three Indian major carps (Labeo rohita, Catla catla and Cirrhinus cirrhosus), belonging to two different weight groups (250 and 500 g), collected from ponds at two different sites (Nalban bheri and Diamond Harbour). The tissues were analysed for the levels of different antioxidant defence systems such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GRd), glutathione S-transferase (GST), glutathione (GSH) and malondialdehyde (MDA). Concentrations of all the metals were significantly higher (P < 0.05) in sediment, water and the tissues from Nalban bheri compared to those in Diamond Harbour. Metal concentrations were the lowest in C. cirrhosus, which increased with an increase in fish weight, and the liver accumulated higher amount of metals than the kidney. Activities of all enzymatic and non-enzymatic antioxidant parameters except GPx and GRd were significantly higher (P < 0.05) in the tissues from Nalban bheri than those in Diamond Harbour. Significant multicollinearity was found in the values of SOD, CAT, GST, GRd, GPx and MDA with Pb, Cu and Ni in all three fish species at Nalban and with Cd in L. rohita and C. catla. Principal component analysis results revealed that stress response in a polluted site was directly regulated by an amalgamation of GSH profile and the levels of MDA in a synchronized manner. The study indicated a tissue-specific and species-specific difference for heavy metal-induced oxidative stress response in fish and a correlation between different heavy metals and individual oxidative stress markers.     

Biomarker responses as indication of contaminant effects in Oreochromis niloticus

The current study investigated oxidative stress parameters (enzymes activities, metallothionein content and lipid peroxidation) in freshwater fish, Oreochromis niloticus, tilapia exposure to Monjolinho River (in 4 months of year: January, April, July and November). One critical site in Monjolinho River (site B) was assessed in comparison to a reference site (site A). Water pH and oxygen concentration was lower than that recommended by CONAMA (Brazilian National Environmental Committee), resolution 357/2005 for protection of aquatic communities, and ammonium and the metals Cu, Zn, Mn and Fe (on all months) concentrations were higher than the maximum concentration recommended. Glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities were significantly decreased in liver and muscle in tilapia from Monjolinho River, throughout the year, in relation to reference except in gills that SOD activity increased. Glutathione S-transferase (GST) activity was significantly increased in liver of the tilapia from Monjolinho River in all sites, in relation to reference except in gills that GST activity increased in July and decreased in November, suggesting that GST activity could be induced to neutralize the pollutants toxicity. On the other hand, GST activity was significantly decreased in white muscle indicating a toxic effect of pollutants, resulting in a decreased ability of tilapia to perform defense reactions associated to GSTs. The decrease of catalase (CAT) activity in gills of the O. niloticus together with the increase of SOD activity, could explain the increased lipid peroxidation (LPO) level in this organ. Metallothionein levels in liver and gills were significantly high in all sites. Results indicate that the exposure to metals caused severe damage to tissues; despite the consensually assumed antioxidant induction as a sign of exposure to contaminants the effects seem in part to be mediated by suppression of antioxidant system with SOD, CAT and GPx as potential candidates for tissues toxicity biomarkers of pollutants.     

Ecotoxicological effects on the earthworm Eisenia fetida following exposure to soil contaminated with imidacloprid

Imidacloprid, a neonicotinoid insecticide, has been used widely in agriculture worldwide. The adverse effects of imidacloprid on exposed biota have brought it increasing attention. However, knowledge about the effects of imidacloprid on antioxidant defense systems and digestive systems in the earthworm is vague and not comprehensive. In the present study, the changes in the activity of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), cellulase, reactive oxygen species (ROS), and malondialdehyde (MDA) in the earthworm Eisenia fetida exposed to artificial soil treated with imidacloprid were examined systematically. The results showed that the activity of these biomarkers was closely related to the dose and duration of the exposure to imidacloprid. The activity of SOD was stimulated significantly at doses of 0.66 and 2 mg kg^?1 imidacloprid but markedly inhibited at a dose of 4 mg kg^?1 imidacloprid with prolonged exposure. The activities of CAT and POD increased irregularly at 0.2–4 mg kg^?1 imidacloprid over different exposure times. The level of ROS at a dose of 2 or 4 mg kg^?1 imidacloprid was significantly increased over the entire exposure period. When the concentration of imidacloprid was above 0.66 mg kg^?1, the balance of the activity of the antioxidant enzymes and ROS level was interrupted. The activity of cellulase decreased significantly with prolonged exposure. At the stress of 4 mg kg^?1 imidacloprid, the content of MDA was significantly increased with increasing exposure time. The results of the present study suggest that imidacloprid has a potentially harmful effect on E. fetida and may be helpful for assessment of the risk of imidacloprid to the soil ecosystem environment. However, to obtain more comprehensive toxicity data, it is necessary to investigate the effects of imidacloprid on earthworm using native soils in the future work.     

A systematic review and meta-analysis of lead and cadmium concentrations in cow milk in Iran and human health risk assessment

The aim of the current research was to systematically review and summarize the studies that evaluated the concentration of lead (Pb) and cadmium (Cd) in cow milk in different regions of Iran and to perform a meta-analysis of the findings. Moreover, the non-carcinogenic and carcinogenic risks of Pb and Cd through milk consumption in adult and child consumers were assessed. As a result of a systematic search in the international and national databases between January 2008 and October 2018, 17 reports involving 1874 samples were incorporated in our study for meta-analysis. The pooled concentrations of Pb and Cd were estimated to be 13.95 μg mL−1 (95% CI 9.72–18.11 μg mL−1) and 3.55 μg mL−1 (95% CI − 2.38–9.48 μg mL−1), respectively, which were lower than the WHO/FAO and national standard limits. The estimated weekly intake (EWI) of Pb and Cd through consuming milk was 16.65 and 7 μg day−1 for adults of 70 kg and 45 and 34 μg day−1 for children of 26 kg, respectively, which was well below the risk values set by Joint FAO/WHO Expert Committee on Food Additives (JECFA). The maximum target hazard quotient values (THQs) of Pb and Cd were 5.55E−5 and 5.55E−5 for adults and 5.55E−5 and 5.55E−5 for children, respectively, which were lower than 1 value, suggesting that Iranian consumers are not exposed to non-carcinogenic risk through consuming milk. Moreover, the incremental lifetime cancer risk (ILCR) of Pb estimated to be 2.96E−04 in adults and 1.0E−03 in children, indicating that consumers in Iran are at threshold carcinogenic risk of Pb through consuming milk (ILCR > 10−4). Therefore, planning and policy making for the sustainable reduction of these toxic metals in milk, particularly in industrial regions of Iran, are crucial.     

Metabolic responses of weeping willows to selenate and selenite

Goal, Scope and Background Selenium (Se) is one of the most widely distributed elements of the earth’s crust at low concentrations. The extensive use of Se-containing chemicals due to anthropogenic activities has increased the ecological risk to environmental compartments. Plants, under unfavorable environmental conditions, often increase the formation of reactive oxygen species (ROS), and consequently plant antioxidant enzymatic systems have been proposed to be important in plant stress tolerance. The goal of this study was to find out the metabolic responses of plants to Se, to provide quantitative information whether exogenous Se has a beneficial role in plants, and to investigate the potential of vegetation management of Se for potential phytoremediation. Material and Methods Pre-rooted plants of weeping willows (Salix babylonica L.) were grown hydroponically in growth chambers and treated with Na₂SeO₄ or Na₂SeO₃ at 24.0 ± 1°C for 168 h. Five different treatment concentrations were used, ranging from 0.44 to 8.72 mg Se/L for the treatments exposed to SeO₄²⁻ and from 0.50 to 10.0 mg Se/L for the treatments exposed to SeO₃²⁻, respectively. Transpiration rates, soluble protein contents and antioxidative enzyme activities of the plants were monitored to evaluate toxicity from exogenous Se exposure. At the end of the study, total Se in the hydroponic solution was analyzed by hydride generation-atomic fluorescence spectrometry (HG-AFS). Results Both chemical forms of Se at low concentrations showed growth-promoting effects on plants. A significant decrease of transpiration rates and of soluble protein contents of plants was observed at higher Se concentrations after 168 h of exposure. Measurable change of superoxide dismutases (SOD) activity in leaves was only detected under high Se treatments. Catalase (CAT) activity was significantly affected by the Se application. Slight change of peroxidase (POD) activity was measured in all treatments, whereas significant inhibition of POD activity was detected for the plants exposed to SeO₃²⁻ of 10.0 mg Se/L. Se-induced stress appeared in all treatments, thus resulting in measurable increase of glutathione peroxidase (GSH-Px) activity of the plants. Although both chemical forms of Se were taken up by weeping willows efficiently, their uptake rates were different. Discussion Of all measured parameters, POD and CAT activities in leaves were noted the most sensitive indicator for the plants exposed to SeO₄²⁻ and SeO₃²⁻, respectively. Deleterious effects on plant physiological functions due to Se application were not observed over 168 h of exposure. This is largely due to the fact that well-established antioxidant enzymatic systems in plants and higher activities of GSH-Px largely reduced the negative effects on plants; SeO₃²⁻ caused much more severe stress to plants than SeO₄²⁻ at higher Se application rates. The uptake mechanisms between the two chemical species were quite different. Conclusions Neither visible toxic symptoms nor metabolic lesions were observed at low concentrations of Se, probably due to the effective established enzymatic systems in weeping willows. All selected parameters for toxicity determination were significantly correlated to Se application, but metabolic responses of plants to SeO₄²⁻ and SeO₃²⁻ were quite different. GSH-Px in leaves was probably the principle enzyme responsible for stress reduction from Se exposure. Due to their different chemical properties, weeping willows showed a faster uptake rate for SeO₄²⁻ than for SeO₃²⁻. Recommendations Exogenous Se has a beneficial role in plants and vegetation management of Se is a potential remediation strategy in cleaning up Se-contaminated sites. Further investigation on the biochemical mechanism of Se metabolism will provide insight to the specific interactions between Se and plants on the molecular level. Perspectives Weeping willow has a sound potential for phytoremediation of Se-contaminated sediment and groundwater because the tree is not only tolerant to Se but also uptakes chemical species from the environment.     

Tracking mercury in the southwestern Atlantic Ocean: the use of tuna and tuna-like species as indicators of bioavailability

Mercury is a trace element that is potentially dangerous due its high toxicity and tendency to bioaccumulate in organisms. Currently, high mercury concentrations are seen in the environment especially due climate changes. Studies regarding mercury bioavailability in the southwestern Atlantic Ocean using tuna and tuna-like species are rare. The aim of the present study was to use tuna and tuna-like species (Thunnus atlanticus, Thunnus albacares, Katsuwonus pelamis, Euthynnus alletteratus, Coryphaena hippurus and Sarda sarda) as indicators of the availability of total mercury (THg) in oceanic food webs of the southwestern Atlantic Ocean. THg concentrations varied significantly among species for both muscle and liver (Kruskal–Wallis test; H5,130 = 52.7; p < 0.05; H5,130 = 50.1; p < 0.05, respectively). The lowest concentrations were found in C. hippurus (0.008 mg kg−1 wet weight in the muscle and 0.003 mg kg−1 wet weight in the liver), and the highest concentrations were reported in the muscle of T. atlanticus (1.3 mg kg−1 wet weight) and in the liver of S. sarda (2.5 mg kg−1 wet weight). The continued monitoring of tuna and tuna-like species is necessary to assist in their conservation since tuna can be sentinels of mercury pollution.     

Influence of UV radiation on chlorophyll,and antioxidant enzymes of wetland plants in different types of constructed wetland

A surface- and vertical subsurface-flow-constructed wetland were designed to study the response of chlorophyll and antioxidant enzymes to elevated UV radiation in three types of wetland plants (Canna indica, Phragmites austrail, and Typha augustifolia). Results showed that (1) chlorophyll content of C. indica, P. austrail, and T. augustifolia in the constructed wetland was significantly lower where UV radiation was increased by 10 and 20 % above ambient solar level than in treatment with ambient solar UV radiation (p?C. indica, P. australis, and T. angustifolia by elevated UV radiation of 10 % was higher in vertical subsurface-flow-constructed wetland than in surface-flow-constructed wetland. The sensitivity of MDA, SOD, POD, and CAT activities of C. indica, P. austrail, and T. augustifolia to the elevated UV radiation was lower in surface-flow-constructed wetland than in the vertical subsurface-flow-constructed wetland, which was related to a reduction in UV radiation intensity through the dissolved organic carbon and suspended matter in the water. C. indica had the highest SOD and POD activities, which implied it is more sensitive to enhanced UV radiation. Therefore, different wetland plants had different antioxidant enzymes by elevated UV radiation, which were more sensitive in vertical subsurface-flow-constructed wetland than in surface-flow-constructed wetland.     

Pharmacological and ameliorative effects of Withania somnifera against cadmium chloride–induced oxidative stress and immune suppression in Nile tilapia,Oreochromis niloticus

This study was carried out to evaluate the effects of dietary supplementation of aqueous extract of Withania somnifera (W. somnifera) against cadmium chloride–induced toxicity in the Nile tilapia, Oreochromis niloticus. Five experimental groups were designed: group (I) was free from cadmium chloride and W. somnifera and served as a control, group (II) was exposed to 1.775 mg L^?1 of cadmium chloride only (which is equivalent to 1/4 96-h LC50), while groups (III), (IV), and (V) were exposed to 1.775 mg cadmium chloride L^?1 with co-supplementation of dietary W. somnifera in doses of 1.0, 2.0, and 3.0 mL kg^?1 body weight (bwt), respectively. The experiment lasted for 4 weeks. In the second and fourth weeks of the experiment, the following indicators were evaluated: hematological (hemogram and blood protein profile), biochemical (activities of serum liver enzymes, namely alanine transaminase (ALT) and aspartate transaminase (AST)), immunological (immunoglobulin M (IgM), serum lysozyme), and tissue antioxidant changes (malondialdehyde (MDA) levels and activities of catalase (CAT) and superoxide dismutase (SOD)). Additionally, gene expressions of glutathione-S-transferase (GST) in the liver were assessed. At the end of the experiment, all fish in all groups were experimentally challenged with Aeromonas hydrophila and the relative protection survival (RPS) was demonstrated. The results revealed that groups exposed to cadmium chloride toxicity and co-supplemented with dietary aqueous extract of W. somnifera at high doses showed significant ameliorative effects in hemogram parameters, total protein, globulin, IgM, and lysozyme against cadmium chloride–induced toxicity compared to the control group and the group exposed to a sublethal dose of cadmium chloride without co-suplemntation of W. somnifera. The results showed also that groups supplemented orally with W. somnifera at high doses have higher antioxidant activities of CAT and SOD and reduction of MDA formation. Levels of gene expressions of GST in the liver were higher in W. somnifera extract-supplemented groups more than those in the group exposed to cadmium chloride–induced toxicity without W. somnifera supplementation. In addition, the results revealed improved RPS with the dietary supply of W. somnifera extract in high doses. In conclusion, this study showed that the dietary supplementation of W. somnifera extract to diets of O. niloticus could be suggested as an effective way to overcome cadmium chloride–induced toxicity because it improves blood parameters and antioxidants, and it can be used as an immunostimulant against the invading bacterial pathogens.

     

Subacute oral toxicity of BDE-15, CDE-15, and HODE-15 in ICR male mice: assessing effects on hepatic oxidative stress and metals status and ascertaining the protective role of vitamin E

The present study examined the effects of oral exposure of 4,4′-dibromodiphenyl ether (BDE-15), 4,4′-dichlorodiphenyl ether (CDE-15), and 4,4′-dihydroxydiphenyl ether (HODE-15) on hepatic oxidative stress (OS) and metal status in Institute of Cancer Research (ICR) male mice. Furthermore, the role of vitamin E in ameliorating potential OS caused by BDE-15, CDE-15, and HODE-15 was investigated. Three groups of mice were exposed to 1.20 mg/kg(body weight)/day of each of the three toxicants for 28 days. Results showed that none of the three toxicants altered growth rates of mice, but significantly increased (P?<?0.05) relative liver weights and decreased relative kidney weights. Pathological changes including cell swelling, inflammation and vacuolization, and hepatocellular hypertrophy in livers were observed. Significant decreases (P?<?0.05 and P?<?0.01) in superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) activity, and glutathione (GSH) levels, together with increases in malondialdehyde (MDA) content were recorded in all toxicant-treated groups. Hepatic copper levels increased in all toxicant-treated groups. Hepatic zinc levels decreased in the liver of BDE-15-treated mice, whereas they increased in the livers of CDE-15-treated and HODE-15-treated mice. In conclusion, daily exposure to the three toxicants perturbed metal homeostasis and increased OS in mouse liver. Experimental data indicated the hepatic oxidative toxicity of the three toxicants followed the order BDE-15?<?HODE-15?<?CDE-15. Moreover, the study proved that daily supplementation of 50 mg/kg vitamin E is effective to ameliorate the hepatic OS status and metal disturbance in mice.     

Ultrasonic vibration seeds showed improved resistance to cadmium and lead in wheat seedling

Heavy metals have long-term adverse impacts on the health of soil ecosystems and even exhibit hazardous influences on human health. Literatures have shown that heavy metals could result in the reduction of crops growth and development and finally result in crops production decline. To determine whether or not ultrasonic vibration alleviate damage induced by cadmium and lead in crops, the wheat seeds, which is one of the most important agriculture crops in China and other countries in the world, were exposed to 10 min ultrasonic vibration and then the toxicological effects were investigated. Wheat seeds were soaked for 3 h with water and then the seeds were placed in clean beaker with some water, the beaker were placed in ultrasonic apparatus to vibrate (model, KQ-200VDV; frequency, 45 KHz; power, 160 W). Pretreatment seeds of 80 were sown in dishes (Ø 15 cm). After seeds emergence, the seedlings were thinned to 60 per dish. The dishes with seedlings were placed in a growth chamber maintained at 25 °C, 70 % relative humidity and 380 μmol?mol^?1 CO₂ under dark condition. A 400 μmol?m^?2?s^?1 photosynthetically active radiation was provided for 8 h (dark for 16 h) after the seed germination. When the seedlings were 2 days old, the seedlings were subjected to cadmium and lead for 4 days and then some selective biochemical and physiological parameters were measured. (1) Although each doses of ultrasonic vibration could improve seed germination, enhance biosynthesis of protein and chlorophyll and seedlings growth, the optimum dosage of ultrasonic vibration was 10 min. (2) Compared with the controls, cadmium and lead stress led to significant increase in the concentrations of malondialdehyde (MDA) and O^?2 and in the conductivity of electrolyte leakage, but the activities of catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GR), the glutathione concentration, and the shoot weight were decreased by Cd and Pb stress. In the case of the seeds exposed to ultrasonic vibration and the seedlings followed by cadmium lead stress, the concentrations of MDA and O^?2, and the conductivity of electrolyte leakage were significantly lower than those in cadmium and lead stress; the activities of CAT, SOD, and GR and the shoot weight were significantly higher (except for glutathione (GSH) concentration) than those in cadmium and lead stress seedlings. The membrane is responsible for the selective inflow and outflow of molecules, ions, and water, and is a dynamic structure that performs a variety of functions. Cellular membrane systems play an important role in the compartmentalization of cells and maintaining intercellular homeostasis. Abiotic and biotic stress can induce functional impairments to the cellular membrane systems through triggering an increased formation of reactive oxygen species (ROS), such as superoxide (O₂ ^?), hydrogen peroxide (H₂O₂), and hydroxyl radicals. There are several pathways that can be utilized to eliminate ROS in plants, e.g., CAT, SOD, and GR and GSH, etc. compared with controls, cadmium, and lead enhanced the concentrations of ROS; decreased the SOD, CAT, and GR activities; the GSH concentration, and the seedling growth. In the case of ultrasonic pretreatment followed by cadmium and lead stresses, the activities of CAT, SOD, and GR were significant higher, and the conductivity of electrolyte leakage and the concentrations of MDA and O₂ ^? were significant lower than that of those subjected by cadmium and lead stress. This phenomenon demonstrated ultrasonic pretreatment can help plant eliminate the ROS by enhance the activities of antioxidant enzymes. These results suggested that ultrasonic vibration can alleviate the toxicological effect induced by heavy mental.     

Effects of di-n-butyl phthalate on the physiology and ultrastructure of cucumber seedling roots

Agricultural pollution caused by the use of plastic sheetings has been documented to be a widespread problem in most of the major crop-planting regions of the world. In order to better understand the phytotoxic mechanisms induced by phthalic acid esters involved with this problem, Cucumber sativus L. cv Jinyan No. 4 were sown in pots to the three-leaf-stage in the presence of di-n-butyl phthalate (DBP; 0, 30, 50, 100, and 200 mg L^?1) for 1, 3, 5, or 7 days. Physiology, biochemistry, and ultrastructure of seedling roots were examined. The results indicated that activities of three antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD)) were stimulated at low-DBP treatments and decreased under higher levels (>100 mg L^?1) compared to the controls. On the other hand, SOD and POD provided a better defense against DBP-induced oxidative damage in the roots of cucumber seeding, compared to CAT. The productions of both malondialdehyde (MDA) and proline (Pro) were promoted under DBP stress. Visible impact on the cytoderm, mitochondrion, and vacuole was detected, possibly as a consequence of free radical generation. These results suggested that activation of the antioxidant system by DBP led to the formation of reactive oxygen species that resulted in cellular damage.     

Assessment of imidacloprid toxicity on reproductive organ system of adult male rats

In the current study it was aimed to investigate the toxicity of low doses of imidacloprid (IMI) on the reproductive organ systems of adult male rats. The treatment groups received 0.5 (IMI-0.5), 2 (IMI-2) or 8 mg IMI/kg body weight by oral gavage (IMI-8) for three months. The deterioration in sperm motility in IMI-8 group and epidydimal sperm concentration in IMI-2 and IMI-8 groups and abnormality in sperm morphology in IMI-8 were significant. The levels of testosterone (T) and GSH decreased significantly in group IMI-8 compared to the control group. Upon treatment with IMI, apoptotic index increased significantly only in germ cells of the seminiferous tubules of IMI-8 group when compared to control. Fragmentation was striking in the seminal DNA from the IMI-8 group, but it was much less obvious in the IMI-2 one. IMI exposure resulted in elevation of all fatty acids analyzed, but the increases were significant only in stearic, oleic, linoleic and arachidonic acids. The ratios of 20:4/20:3 and 20:4/18:2 were decreased and 16:1n-9/16:0 ratio was increased. In conclusion, the present animal experiments revealed that the treatment with IMI at NOAEL dose-levels caused deterioration in sperm parameters, decreased T level, increased apoptosis of germ cells, seminal DNA fragmentation, the depletion of antioxidants and change in disturbance of fatty acid composition. All these changes indicate the suppression of testicular function.     

Assessment of Jatropha curcas L. biodiesel seed cake toxicity using the zebrafish (Danio rerio) embryo toxicity (ZFET) test

Consequent to the growing demand for alternative sources of energy, the seeds from Jatropha curcas remain to be the favorite for biodiesel production. However, a significant volume of the residual organic mass (seed cake) is produced during the extraction process, which raises concerns on safe waste disposal. In the present study, we assessed the toxicity of J. curcas seed cake using the zebrafish (Danio rerio) embryotoxicity test. Within 1-h post-fertilization (hpf), the fertilized eggs were exposed to five mass concentrations of J. curcas seed cake and were followed through 24, 48, and 72 hpf. Toxicity was evaluated based on lethal endpoints induced on zebrafish embryos namely egg coagulation, non-formation of somites, and non-detachment of tail. The lowest concentration tested, 1 g/L, was not able to elicit toxicity on embryos whereas 100 % mortality (based also on lethal endpoints) was recorded at the highest concentration at 2.15 g/L. The computed LC50 for the J. curcas seed cake was 1.61 g/L. No further increase in mortality was observed in the succeeding time points (48 and 72 hpf) indicating that J. curcas seed cake exerted acute toxicity on zebrafish embryos. Sublethal endpoints (yolk sac and pericardial edema) were noted at 72 hpf in zebrafish embryos exposed to higher concentrations. The observed lethal endpoints induced on zebrafish embryos were discussed in relation to the active principles, notably, phorbol esters that have remained in the seed cake even after extraction.     

Assessment of imidacloprid toxicity on reproductive organ system of adult male rats

In the current study it was aimed to investigate the toxicity of low doses of imidacloprid (IMI) on the reproductive organ systems of adult male rats. The treatment groups received 0.5 (IMI-0.5), 2 (IMI-2) or 8 mg IMI/kg body weight by oral gavage (IMI-8) for three months. The deterioration in sperm motility in IMI-8 group and epidydimal sperm concentration in IMI-2 and IMI-8 groups and abnormality in sperm morphology in IMI-8 were significant. The levels of testosterone (T) and GSH decreased significantly in group IMI-8 compared to the control group. Upon treatment with IMI, apoptotic index increased significantly only in germ cells of the seminiferous tubules of IMI-8 group when compared to control. Fragmentation was striking in the seminal DNA from the IMI-8 group, but it was much less obvious in the IMI-2 one. IMI exposure resulted in elevation of all fatty acids analyzed, but the increases were significant only in stearic, oleic, linoleic and arachidonic acids. The ratios of 20:4/20:3 and 20:4/18:2 were decreased and 16:1n-9/16:0 ratio was increased. In conclusion, the present animal experiments revealed that the treatment with IMI at NOAEL dose-levels caused deterioration in sperm parameters, decreased T level, increased apoptosis of germ cells, seminal DNA fragmentation, the depletion of antioxidants and change in disturbance of fatty acid composition. All these changes indicate the suppression of testicular function.     

Lead induced changes in antioxidant metabolism of horsegram (Macrotyloma uniflorum (Lam.) Verdc.) and bengalgram (Cicer arietinum L.)

One-month old horsegram (Macrotyloma uniflorum (Lam.) Verdc. cv VZM1) and bengalgram (Cicer arietinum L. cv Annogiri) were exposed to different regimes of lead stress as Pb(NO3)2 at 0, 200, 500 and 800 ppm concentrations. The extent of oxidative damage as the rate of lipid peroxidation, antioxidative response and the accumulation of lead in roots and shoots of both plants were evaluated after 12 days of lead stress. Lead (Pb) treated plants showed increased levels of lipid peroxidation as evidenced from the increased malondialdehyde content coupled with the increase in the activities of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), glutathione reductase (GR), glutathione S-transferase (GST) compared to control (untreated) plants. Lead stress caused significant changes in the activity of antioxidative enzymes. The effect of lead was found to be concentration dependent. Higher concentration of lead (800 ppm) resulted 2- to 3-fold increase in SOD, catalase and peroxidase activities, 3- to 5-fold increase in GR activity and 3- to 4-fold increase in GST activity in roots and leaves of both horsegram and bengalgram plants. Lead stress caused a significant increase in the rate of peroxidation as showed in the levels of malondialdehyde content in roots and leaves of both plant species. Horsegram registered lower Pb accumulation than bengalgram, however localization of Pb was greater in roots than leaves in both plants. In general, lipid peroxide levels and antioxidative enzyme activities were higher in horsegram than bengalgram and also more in roots than leaves which best concordance with the lead contents of both the plants and organs. These results suggest that Pb toxicity causes oxidative stress in plants and the antioxidative enzymes SOD, CAT, POD, GR, GST could play a pivotal role against oxidative injury.