Agnieszka Zawisza-Raszka

Accumulation of heavy metals and antioxidant responses in Vicia faba plants grown on monometallic contaminated soil

The purpose of this study was to explore the effects of soil contamination by selected metals (cadmium, copper, nickel, lead or zinc) on the antioxidant response of Vicia faba plants. The levels of the antioxidants: glutathione, proline, non-protein thiols, as well as guaiacol peroxidase and catalase activities were measured in the upperparts of plants. Additionally, the potential bioavailability of metals in the soil and their concentrations in V. faba plants were compared. Treatment with metal caused the problem of an elevation in its bioavailability in soil and its concentration in leaves and stems. The most serious problems seemed to be metal elevations in soil, especially Zn and Ni as well as in the aerial parts of V. faba plants. The antioxidant responses appeared to be metal specific. The elevation of guaiacol peroxidase activity in leaves and stems as well as the proline in leaves was the only more general reaction to metal exposure. Upon analysis of the effects of soil metal contamination on V. faba plants, we recommend the use of some measurements such as guaiacol peroxidase activity and proline level as useful tools in biological monitoring.

Effects of multigenerational cadmium exposure of insects (Spodoptera exigua larvae) on anti-oxidant response in haemolymph and developmental parameters

Biochemical and organismal indices of metal tolerance were studied in Spodoptera exigua exposed to a cadmium-contaminated diet for one or many (33 or 61) generations. Reduced and oxidised glutathione, protein thiols, total anti-oxidant capacity level, glutathione transferase activity, and Cd accumulation were assayed in the haemolymph of the last instar larvae. The cadmium concentration in the whole larval body as well as larval survival, larval duration time and last instar body weight were also measured. Elevated cadmium concentration in the whole body, higher mortality and longer duration of the larval stage in one-generation exposed insects in comparison with those exposed for many generations suggest that metal tolerance builds over time. For the larvae from multigeneration metal treatment, the higher cadmium concentration in larval haemolymph positively correlated with glutathione oxidation and total anti-oxidant capacity. One-generation exposed insects had lower metal concentration in haemolymph than did 33-generation exposed insects.

The effects of cadmium or zinc multigenerational exposure on metal tolerance of Spodoptera exigua (Lepidoptera: Noctuidae)

The effects of ten generational zinc or cadmium pre-exposure on metal tolerance among beet armyworm Spodoptera exigua individuals were compared. These effects were assessed in animals from the 11th generation, reared on a diet either uncontaminated or contaminated with metal (cadmium or zinc). The survival rate of larvae and the degree of metal accumulation (in larvae, pupae and moths; among larval organs: gut and fat body) were analysed. Catalase, superoxide dismutase and glutathione transferase activity in larval organs of individuals subjected to different metal treatments were also measured. Animals transferred from control rearing to metals (cadmium or zinc) in the 11th generation, as well as those from multigenerational zinc treatment, but not from multigenerational cadmium treatment, had a significantly lower survival rate than control animals. Insects from the groups with the high metal treatment had high bioaccumulation factors (above 3.7 and 2.3 following cadmium and zinc, respectively). Cadmium (but not zinc) pre-exposure had a significant effect on metal accumulation in larvae. Multigenerational metal pre-exposure seemed to have mainly a negative effect on glutathione transferase activity in the gut of larvae from the 11th generation, in the case of the individuals exposed to metal other than that used in pre-exposure treatment or kept in control conditions. However, in the case of zinc pre-exposure, such effect was only apparent when zinc was replaced by cadmium. The long-term effect of cadmium on catalase activity in larvae was found.

Effects of nickel exposure and acute pesticide intoxication on acetylcholinesterase, catalase and glutathione S-transferase activity and glucose absorption in the digestive tract of Helix aspersa (Pulmonata, Helicidae).

The activity of detoxifying enzymes and glucose absorption were measured in the digestive tract of Helix aspersa exposed to nickel. Nickel decreased CAT activity and, in the lower concentration, inhibited intestinal absorption of glucose. Elevated AChE activity and unchanged GST activity indicate diversity in enzymatic response to nickel. Nickel at higher concentration, destroying the intestinal wall integrity, caused high glucose influx. Nickel pre-treatment augmented the response to a single diazinon application. AChE activity was greatly reduced compared with nickel-untreated snails. The reduction in CAT activity was similar in both groups. Glucose absorption remained unchanged.

DNA damage in grasshopper Chorthippus brunneus (Orthoptera) hatchlings following paraquat exposure

Comet assay was applied to study genotoxic damage induced by paraquat (PQ) in brain cells of Chorthippus brunneus (Insecta: Orthoptera) hatchlings. Percentage of the comet fluorescence in the tail (TDNA), length of the comet tail (TL) and Olive tail moment (OTM) were used for quantitative assessment of the DNA damage. Multiple regression analysis supplemented standard statistical elaboration of the results. Increasing PQ concentrations applied either directly to the brain cells suspension (10, 50, and 250 μM PQ final concentration--in vitro protocol) or indirectly (50, 250, and 1250 μM PQ final concentration--in vivo protocol) provoked significant increase of oxidative damage to DNA (higher median TDNA and OTM values). The damage increased with time of exposure (0, 5, 15, and 30 min) following in vitro application, but decreased in longer interval (3 vs 24 h) after in vivo administration of paraquat. On contrary, median TL values did not correlate with paraquat concentration irrespectively of the exposure protocol. Possible reason of this discrepancy in light of paraquat toxicity is discussed.

ACETYLCHOLINESTERASE, CATALASE AND GLUTATHIONE S-TRANSFERASE ACTIVITY IN BEET ARMYWORM (SPODOPTERA EXIGUA) EXPOSED TO NICKEL AND/OR DIAZINON

The effect of single and combined action of nickel and pesticide (diazinon) on enzymes activity (glutathione S-transferase, catalase and acetylcholinesterase) in the digestive tract, body wall and fat body as well as basic growth parameters (life span and body mass) of Spodoptera exigua were investigated under laboratory conditions. The experiment was carried out on two nickel treated groups [300 (NiI) and 900 (NiII) mg Ni kg(-1) dry weight of the culture medium] and a control group. The results showed that mortality of caterpillars in NiII group was higher (51.1%) when compare with the controls. The body mass of the caterpillars in the NiI group was higher by 20% than in the control group, and the body mass of the pupae in the NiII group was lower by 22% than in the control group. Exposure to nickel influenced AChE, GST and catalase activity in the body wall (increase up to 66%) and GI tract, while in the fat body the above-mentioned activity remained unchanged. The pesticide application caused a strong, about 70% reduction in AChE activity in GI tract, while in case of GST activity pesticide treatment resulted in multidirectional response depending on the organ. Nickel pre-treatment affects the susceptibility to pesticide, which is manifested in a lower activity of GST and catalase in the fat body (from 26 to 36%), when compare with the other experimental groups.

Phenotypic Plasticity, Epigenetic or Genetic Modifications in Relation to the Duration of Cd-Exposure within a Microevolution Time Range in the Beet Armyworm

In the case of the pests inhabiting metal polluted or fields where the use of pesticides is common, a natural selection of resistant individuals can occur. This may pose serious problems for humans, agriculture, as well as the economies of many countries. In this study, the hypothesis that multigenerational (120 generations) exposure to cadmium of a beet armyworm population could be a selecting factor toward a more efficient DNA protection was verified. The hemocytes of individuals from two culture strains (control and Cd-exposed) were treated with H2O2 (a DNA-damaging agent) or PBS (reference). The level of DNA damage was assessed using the Comet assay immediately and 5, 15 and 30 min. after the treatment. The immediate result of the contact with H2O2 was that the level of DNA damage in the hemocytes of the insects from both strains increased significantly. However, in the cells of the Cd-exposed individuals, the level of DNA damage decreased over time, while in the cells from the control insects it remained at the same level with no evidence of repair. These results suggest that efficient defense mechanisms may exist in the cells of insects that have prolonged contact with cadmium. Some evolutionary and trade-off aspects of the phenomenon are discussed. In a wider context, comparing the results obtained in the laboratory with field studies may be beneficial for understanding basic mechanisms of the resistance of an organism. To summarize, the high potential for the repair of DNA damage that was observed in the insects from the cadmium strain may confirm the hypothesis that multigenerational exposure to that metal may possibly contribute to the selection of insects that have a wider tolerance to oxidative stress. However, our investigations of polymorphism using AFLP did not reveal differences between the two main insect strains.

Cross tolerance in beet armyworm: long-term selection by cadmium broadens tolerance to other stressors

Long lasting exposure of animals to stressing factor may lead to the selection of population able to cope with the stressor at lower cost than unexposed individuals. The aim of this study was to assess whether 130-generational selection of a beet armyworm to cadmium in food might have induced tolerance also to other stressors. The potential tolerance was assessed by means of unspecific stress markers: HSP70 concentration, DNA damage level, and energy budget indices in L5 larval instars of beet armyworm. The animals originated from Cd-exposed and control strains exposed additionally in a short-term experiment to high/low temperature or pesticide—spinosad. The application of the additional stressors caused, in general, an increase in the levels of studied parameters, in a strain-dependent manner. The most significant increase was found in HSP70 level in the individuals from the Cd-strain exposed to various spinosad concentration. Therefore, multigenerational contact with cadmium caused several changes that enable the insect to survive under a chronic stress, preparing the organism to the contact with an additional, new stressor. This relationship may be described as a sort of cross tolerance. This may, possibly, increase the probability of population survivorship and, at the same time, decrease the efficiency of pesticide-based plant protection efforts.

Ileal transposition in rats influenced glucosemetabolism and HSP70 levels

Abstract Objective: Ileal transposition procedure (IT), in combination with sleeve gastrectomy, is widely used to induce diabetes remission and to control related metabolic abnormalities. A transposition of a long segment of distal ileum in obese Zucker rats improved glucose tolerance 6 months after IT. The premise of our study was to to examine the long - term effects of ileum transposition on the liver glycolytic enzymes content in a euglycemic group of operated Zucker rats. Methods: Twenty male Zucker rats underwent either the transposition of 50% distal ileum or a sham surgery. Six months after surgery, liver tissue concentrations of glycogen synthase kinase alpha (GSK-3α), glucose 6-phosphatase (G6PC), glycogen phosphorylase (PYGM) and phosphofructokinase (PFK) and HSP70 were assessed by immunoenzymatic methods. Results: HSP70 values were significantly higher in the IT group compared to SHAM. G6PC liver concentrations in the IT group were almost 1.45-fold lower than in the SHAM operated rats. Statistical analyses (F-test) showed HSP70 levels were significantly related to caveolin-1and SHAM group. Conclusions: Lowered glycolytic enzyme concentrations assessed in the liver suggest positive effects on glucose metabolism in long-term observations.

Nickel in Soil Modifies Sensitivity to Diazinon Measured by the Activity of Acetylcholinesterase, Catalase, and Glutathione S-Transferase in Earthworm Eisenia fetida

Nickel in typical soils is present in a very low concentration, but in the contaminated soils it occurs in locally elevated concentrations. The aim of this study was to examine the effect of nickel in the concentrations of 300 (very high, close to LOEC for reproduction) and 900 (extremely high, close to LOEC for mortality) mg/kg dry soil on the life history and acetylcholinesterase, catalase, and glutathione S-transferase activities in earthworm Eisenia fetida and to establish how nickel modifies the sensitivity to organophosphorous pesticide—diazinon. Cocoons production and juveniles’ number were significantly lower only in groups exposed to Ni in the concentration of 900 mg/kg dry soil for two months. Diazinon administration diminished the AChE activity in the GI tract and in the body wall. The interaction between diazinon and nickel was observed, and, in consequence, the AChE activity after the pesticide treatment was similar to controls in worms preexposed to nickel. Both pesticide administration and exposure to nickel caused an increase in the GST activity in examined organs and CAT activity in body wall. Both biometric and development data and simple enzymatic analysis, especially the AChE and GST, show a Ni pretreatment effect on the subsequent susceptibility to pesticide.