Adaptation by death? Cell death-based tolerance to cadmium in 150-generation exposure of Spodoptera exiqua Hübner (Lepidoptera: Noctuidae).

Mechanisms, including autophagy and apoptosis, which serve to regulate and ensure proper organism functions under optimal conditions, play additional defensive roles under environmental pressure. The aim of this study was to test the following hypotheses: (i) elevated autophagy and apoptosis intensity levels, as defensive processes in response to contact with cadmium, are maintained for a limited number of generations and (ii) the number of generations after which levels of cell death processes reach the reference level depends on selective pressure. Cell death processes were assessed by light and transmission electron microscopy, terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL), and cytometric analyses. Model insects (Spodoptera exiqua, Hübner, 1808) were orally exposed to various concentrations of cadmium for 18 generations and compared with reference strains exposed to cadmium or not (control) for over 150 generations. Elevated programmed cell death intensity levels decreased after several generations, indicating tolerance of individuals to cadmium in the diet and verifying the first hypothesis; however, testing the second hypothesis indicated that the number of generations depended not only on pressure intensity, but also on cell death type, since levels of autophagy remained increased for a minimum of 12 generations.

Joint cadmium and polypropylene microparticle action in cadmium tolerant model insect.

Microplastic enlisted as a contaminant of emerging concerns in polluted environments interact with "traditional" contaminants such as metals, causing, among others, their increased accumulation in the body. Harmful effects depend on the exposed animals' possible preadaptation and/or cross-tolerance. The project aimed to assess the role of this phenomenon in the limited toxicity of polypropylene fibers (PPf) in 0%, 0.02%, 0.06, 0.18%, 0.54%, and 1.6% of Cd-supplemented food of larvae of Spodoptera exigua multigenerationally selected to cadmium tolerance. The activity of 20 digestive enzymes (API-ZYM test), defensins, and heat shock proteins, HSP70 levels in the exposed groups were used as biomarkers. PPfs caused the increase of Cd accumulation in the body, while intake of polypropylene microfibers did not change the biomarker levels. Moreover, multigenerational Cd pre-exposure, due to increased tolerance of Cd and, possibly, cross-tolerance, prepares the insects for an additional stressor (PPf) alone and in interaction with cadmium.

Starvation and cadmium affect energy reserves and oxidative stress in individuals of Spodoptera exigua.

Different factors, such as starvation and metal exposure, may affect development and cause oxidative stress in insects. Some host plants may contain a high concentration of cadmium due to their hyperaccumulating property. The negative effects of metals and hunger may be manifested by low availability of energetic substrates. This study aimed to assess whether the insect population with a history of long metal exposure may better manage metal stress or/and starvation at different developmental stages, with the use of energetic substrates. Two strains of Spodoptera exigua model organism were tested: control strain and cadmium strain (treated continuously for over 200 generations with subtoxic amounts of cadmium). The effects of different factors, individually and in combination, on the tested strains were assessed, first by determining the body weight of larvae and pupae and then by estimating the concentration of biomolecules (proteins, carbohydrates, lipids, or glycogen) in the 4th and 5th larval stages and in pupae, and the total antioxidant capacity and lipid peroxidation level in the 4th larval stage. Compared to control strain, cadmium strain individuals exhibited changes in the concentration of soluble carbohydrates and protein. This was partly related to earlier 1-day starvation. In particular, changes in carbohydrate concentration seemed to be a sensitive biomarker of metal stress, independent of the age of individuals and period of starvation. However, the increase in the total antioxidant capacity and the concentration of lipid peroxidation products in the 4th larval stage under the effect of cadmium was dependent on strain origin.

Cardioprotective Effect of Acetylsalicylic Acid in the Myocardial Ischemia-Reperfusion Model on Oxidative Stress Markers Levels in Heart Muscle and Serum.

Heart failure occurs in increased oxidative stress conditions, which contribute to the progression of pathological changes. Orally or intravenously administered acetylsalicylic acid (ASA, aspirin) is typically used in human patients with acute myocardial ischemia. The study used an experimental porcine ischemia-reperfusion model to evaluate the potential cardioprotective effect of intracoronary administered ASA on myocardial ischemia-reperfusion injury. The cardioprotective effect of ASA was evaluated by measuring selected oxidative stress markers levels in infarcted and non-infarcted myocardium 14 days after the procedure, and three times in serum, before the procedure, during the reperfusion process, and after 14-day recovery. The results showed that intracoronary administrated ASA reduced the oxidative stress. The level of oxidative stress, measured with the non-enzymatic markers total antioxidant capacity (TAC), total oxidative status (TOS), and malondialdehyde (MDA), and the enzymatic markers glutathione peroxidase (GPx), glutathione reductase (GR), and glutathione S-transferase (GST), in heart tissue was significantly higher in a control group injected with saline. The level of oxidative stress in serum, measured with TAC, TOS, oxidative stress index (OSI), and lipofuscin (LF), was also higher in the control group than in animals injected with ASA. The confirmed cardioprotective effect of intracoronary administered ASA provides the foundation for further studies on ASA intracoronary application, which may lead to the development of a new therapy for the treatment of ischemia-reperfusion complications in humans.

Level of oxidative stress for the land snail Cepaea nemoralis from aged and bioremediated soil contaminated with petroleum products.

Here, we investigated whether the widely distributed snail Cepaea nemoralis could be used as a suitable sentinel animal for assessing the effects of soil contaminants-petroleum oil derivatives-after years of soil ageing and treatment with a bacterial formulation. Oxidative stress was assessed in the foot and hepatopancreas of C. nemoralis L. exposed to soil contaminated with unleaded petrol, spent engine oil or diesel oil and bioremediated with a bacterial formulation (soil was used 2 years after contamination and bioremediation process). We measured total antioxidant capacity, catalase and glutathione transferase activity and concentrations of superoxide anions, hydrogen peroxide and protein carbonyls in the foot and hepatopancreas of snails after 2 and 4 weeks of treatment. The studied antioxidant responses appeared largely to be tissue and remediation process specific, while the concentrations of superoxide anions, hydrogen peroxide and protein carbonyls depended on time of exposure, tissue type and the type of contaminants, but mostly not on the remediation process. Generally, changes in the concentrations of superoxide anions, hydrogen peroxide and protein carbonyls in the hepatopancreas of snails seemed to be a suitable measure to assess the risk of animals exposed to soil contaminated with petroleum substances and used after many years of ageing and treatment with a microbial formulation.

Energy reserves, oxidative stress and development traits of Spodoptera exigua Hübner individuals from cadmium strain.

Cadmium as a common environmental stressor may exert highly toxic effects on herbivorous insects. The question was whether possible elevation of an oxidative stress and imbalance of energetic reserves in insects may depend on developmental stage, sex and insect population's multigenerational history of exposure to cadmium. So, the aim of this study was to compare of the development traits, total antioxidant capacity, lipid peroxidation, RSSR to RSH ratio and the concentration of carbohydrates, glycogen, lipids and proteins in whole individuals (larvae or pupae) of Spodoptera exigua originating from two strains: control and selected over 120 generations with sublethal metal concentration (44 Cd mg per dry weight of diet). Generally, the increase of the protein, carbohydrates, glycogen concentration and lipid peroxidation decrease with age of the larvae were found. Revealed cases of a higher mobilisation of carbohydrates and proteins, and changes in total antioxidant capacity or lipid peroxidation, in individuals being under metal exposure, occurred in strain-depended mode. Short-term Cd exposure effect was connected with possible higher engagement of proteins and glycogen in detoxification processes, but also higher concentration of lipid peroxidation. In turn, for long-term Cd exposure effect lower lipids concentration and higher thiols usage seemed to be more specific.

Set of stress biomarkers as a practical tool in the assessment of multistress effect using honeybees from urban and rural areas as a model organism: a pilot study.

A decrease among honey bee populations (Apis mellifera) in the traditional apiaries has been observed in recent years. In light of this negative phenomenon, urban beekeeping seems to be an appropriate alternative solution for the bee population in reducing the toxic effects of a large number of pesticides that are commonly used in agricultural ecosystems. Despite the rapid development of urban beekeeping, there is little information regarding the different aspects of the defense effectiveness of bees from the urban and rural areas. The study was aimed to show whether honey bees from these two locations differ in the level of the valuable biomarkers of stress exposure helpful in establishing which bees, from urban or rural areas, are under greater environmental pressure. For this purpose, foragers from an urban rooftop apiary and a traditional rural apiary were collected. The chosen biomarkers were measured in various tissues of bees. The activity of glutathione S-transferase and acetylcholinesterase, the level of total antioxidant capacity, heat shock protein 70 (Hsp70), and defensin were selected for the analyses. In our opinion, the Hsp70 and defensin levels seemed to be important in the indication of urban multistress factors. The higher level of heat shock proteins and defensins in tissues/organs of bees from the urban apiary-in the gut (an increase, respectively, 92% and 7.3%) and fat body (an increase, respectively, 130% and 7.8%), known as targets of environmental toxins, pointed out the urban environment as highly stressful at both the individual and colony levels. In turn, high total antioxidant capacity was measured in the guts of honey bees from rural area (an increase 107%). Such a situation suggests a different mechanism of defense and specificity of rural and urban environmental stressors and also honey bees foraging activity.

Autophagy: a necessary defense against extreme cadmium intoxication in a multigenerational 2D experiment.

Autophagy is a natural process that aims to eliminate malfunctioning cell parts, organelles or molecules under physiological conditions. It is also induced in response to infection, starvation or oxidative stress to provide energy in case of an energy deficit. The aim of this 2-dimensional study was to test if, and if so, how, this process depends on the concentration of cadmium in food (with Cd concentrations from 0 to 352 µg of Cd per g of food (dry weight)-D1 dimension) and the history of selection pressure (160 vs 20 generations of exposure to Cd-D2 dimension). For the study, the 5th instar larvae of a unique strain of the moth Spodoptera exigua that was selected for cadmium tolerance for 160 generations (44 µg of Cd per g of food (dry weight)), as well as 20-generation (11, 22 and 44 µg of Cd per g of food (dry weight)) and control strains, were used. Autophagy intensity was measured by means of flow cytometry and compared with life history parameters: survivability and duration of the 3rd larval stage. The highest values of autophagy markers were found in the groups exposed to the highest Cd concentration and corresponded (with a significant correlation coefficient) to an increased development duration or decreased survivorship in the respective groups. In conclusion, autophagy is probably initiated only if any other defense mechanisms, e.g., antioxidative mechanisms, are not efficient. Moreover, in individuals from pre-exposed populations, the intensity of autophagy is lower.

DNA damage in Spodoptera exigua after multigenerational cadmium exposure - A trade-off between genome stability and adaptation.

Human activity is a serious cause of extensive changes in the environment and a constant reason for the emergence of new stress factors. Thus, to survive and reproduce, organisms must constantly implement a program of adaptation to continuously changing conditions. The research presented here is focused on tracking slow changes occurring in Spodoptera exigua (Lepidoptera: Noctuidae) caused by multigenerational exposure to sub-lethal cadmium doses. The insects received food containing cadmium at concentrations of 5, 11, 22 and 44 µg per g of dry mass of food. The level of DNA stability was monitored by a comet assay in subsequent generations up to the 36th generation. In the first three generations, the level of DNA damage was high, especially in the groups receiving higher doses of cadmium in the diet. In the fourth generation, a significant reduction in the level of DNA damage was observed, which could indicate that the desired stability of the genome was achieved. Surprisingly, however, in subsequent generations, an alternating increase and decrease was found in DNA stability. The observed cycles of changing DNA stability were longer lasting in insects consuming food with a lower Cd content. Thus, a transient reduction in genome stability can be perceived as an opportunity to increase the number of genotypes that undergo selection. This phenomenon occurs faster if the severity of the stress factor is high but is low enough to allow the population to survive.

Protective role of zinc in Spodoptera exigua larvae under 135-generational cadmium exposure.

The aim of this study was to investigate whether zinc supplementation modulates cadmium toxicity in the beet armyworm Spodoptera exigua selected for 135 generations towards cadmium tolerance. To achieve this, larvae originating from three laboratory populations of S. exigua (control strain - C; cadmium-intoxicated for 135 generations strain - Cd, and control strain intoxicated with Cd for 1 generation - CCd) were additionally exposed to zinc in three concentrations (Zn1, 400 µg Zn·g-1 dry mass of food; Zn2; 200 µg Zn·g-1 dry mass of food; Zn3, 100 µg Zn·g-1 dry mass of food). As the markers of toxicity, a life history traits (the duration of L4 and L5 stages), cellular (DNA damage indices) and biochemical parameters (ADP/ATP ratio and ATP and HSP70 concentrations) were chosen. The duration of larval stages of Zn supplemented larvae was prolonged, while cellular and biochemical indicators, in general, appeared to be lower in comparison to the insects from respective reference groups in each laboratory populations. Moreover, the range of the differences depended on zinc concentration in food. We can suspect that zinc supplementation contributed to the protection of S. exigua individuals against negative effects of cadmium intoxication, probably at the cost of growth rate. Significant differences in the response pattern between insects from different laboratory populations indicate that the influence of additional stress factors is dependent on the overall condition of animals and their previous adaptation to other stressors.

Microevolution or wide tolerance? Level of stress proteins in the beet armyworm Spodoptera eqigua hübner (Lepidoptera: Noctuidae) exposed to cadmium for over 150 generations.

The aim of this study was to investigate whether the cadmium tolerance developed in the beet armyworm Spodoptera exigua selected for over 150 generations may be related to synthesis of the stress proteins metallothioneins (Mts) and 70 kDa heat shock proteins (HSP70). To achieve this, six S. exigua strains (control, k), 150-generation Cd exposure strain (cd), and four 18-generation Cd exposure strains differing in Cd concentration (cd44, cd22, cd11, cd5) were reared. Stress protein level was measured in the midgut of the 5th larval stage after 1-6, 12 and 18 generations. Cd contents was measured in the pupae. Unlike Cd concentration, which depended on metal contents in food but was not generation-dependent, the pattern of Mts and HSP70 concentrations changed in experimental strains from generation to generation. Stress protein levels in the insects exposed to the highest Cd concentration (the same as in the 150-generation Cd exposure strain), initially higher than in the control strain, after the 12th generation did not differ from the level measured in the control strains. It seems therefore that stress proteins play a protective role in insects of lower tolerance to cadmium. The tolerance developed during multigenerational exposure probably relies on mechanisms other than Mt and HSP70 synthesis.

Reproduction and development of Spodoptera exigua from cadmium and control strains under differentiated cadmium stress.

The growth and development of living organisms is programmed in genes, but exogenous factors (e.g. cadmium) may modulate endogenous information. Heavy metals may disturb physiological functions and accumulate in the tissues. The insects under prolonged heavy metal stress show some modifications in their metabolism management. The aim of this study was to compare the reproduction and development between individuals of S. exigua from the strain, exposed over 130 generations to sublethal concentration of cadmium (44 mg Cd/kg dry weight of larval diet), and the individuals from the control strain, both additionally exposed to different concentration of cadmium (22-704 mg Cd/kg dry weight of larval diet). The exposure to various cadmium concentrations in the diet revealed survival difference between the cadmium and the control animals at the larvae stage. The differences between adults were not evident. The telomere length (responsible for the duration of a lifespan) in the cadmium strain was shorter in the females than in the males and the individuals from the control strain. TERF1 gene expression (indirectly responsible for the telomere length) was higher in the individuals from the cadmium strain 24 hrs after eclosion. The significant reduction in the larvae body mass was observed in both strains, when the metal concentration was equal to or higher than 264 mg/kg dry weight of larval diet. The EC50 values (defined as of body mass loss), calculated 48 hours after cadmium exposure of individuals from control and cadmium strains, were respectively 632 and 725 mg Cd/kg dry weight of diet. However, some difference in reproduction (the total number of eggs laid and the oviposition time) between the strains appeared only in the groups fed on the uncontaminated diet. The control females laid almost two times more eggs than those from the cadmium strain, and the control ones had more than two times longer oviposition time than the females from the cadmium strain. The fluctuation was also noted in the size of eggs and the hatching success on the following days when both strains were compared, while the hatching success was higher for the insects from the cadmium strain. In conclusion, the insects from the cadmium strain are more resistant to cadmium contamination, as it is evidenced by the EC50 parameter. However, the females from the cadmium strain start laying eggs statistically later, have shorter telomeres and slightly reduced TERF1 gene expression, but hutching success in the strain is significantly higher when compared with the control individuals.

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.

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.

Different Heavy Metal Accumulation Strategies of Epilithic Lichens Colonising Artificial Post-Smelting Wastes.

Lichens appear to be essential and effective colonisers of bare substrates including the extremely contaminated wastes of slag dumps. This study examines the metal accumulation capacity of epilithic lichens growing directly on the surface of artificial slag sinters. Four species representing different growth forms, i.e., crustose Candelariella aurella, Lecanora muralis, and Lecidea fuscoatra and fruticose Stereocaulon nanodes, were selected to evaluate the relationships between zinc, lead, cadmium, and nickel contents in their thalli and host substrates. Bioaccumulation factors of examined crustose lichens showed their propensity to hyperaccumulate heavy metals. Contrarily, concentrations of metals in fruticose thalli of S. nanodes were, as a rule, lower than in the corresponding substrates. This indicates that the growth form of thalli and degree of thallus adhesion to the substrate has a significant impact on metal concentrations in lichens colonising post-smelting wastes. Nonlinear regression models described by power functions show that at greater levels of Pb concentration in the substrate, the ability of C. aurella, L. muralis and L. fuscoatra to accumulate the metal experiences a relative decrease, whereas hyperbolic function describes a similar trend in relation to Ni content in S. nanodes. This phenomenon may be an important attribute of lichens that facilitates their colonisation of the surface of slag wastes.

Oribatid mite communities on lichens in heavily contaminated post-smelting dumps.

In this study, we examined oribatid fauna of strongly contaminated post-smelting dumps (southern Poland) that exist in the substrate and are associated with the most frequent lichen, Cladonia rei. Due to artificial origin of the substrate and the extremely high contamination with heavy metals, the studied dumps are unique in Europe in terms of unfavourable life conditions. In total, 2,936 specimens of Oribatida, representing 50 oribatid species, were sampled on 10 dumps and a reference site. Thalli of C. rei act as an island for soil oribatid mites on extremely contaminated post-smelting dumps. Both abundance and species richness of oribatid fauna collected from C. rei thalli were significantly greater than those recorded in the dump's substrate. The pool of oribatid species that was able to persist in extremely high doses of heavy metals was comparatively broad. However, only one species, Tectocepheus velatus, was able to achieve high abundances on all dumps. Three different responses of species (tolerant, sensitive, and indifferent) to heavy-metal contamination were recognised. Redundancy analysis indicated that highly increased levels of heavy metals, as well as K content, C/N ratio, and pH value, were the main factors that influence the composition and distribution of species. The concentrations of heavy metals (both essential elements (zinc) as well as xenobiotics (lead, cadmium) in T. velatus from the most contaminated dumps were not increased compared with those observed in moderately contaminated soils.

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.

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 upper parts 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 localisation of HSP70 and oxidative stress indices in heads of Spodoptera exigua larvae in a cadmium-exposed population.

The effects of cadmium toxicity may vary between animals with different history of metal exposure. The aim of our study was to examine HSP70, protein carbonyl levels, catalase activity and total antioxidant capacity in the heads of Spodoptera exigua (Hübner) larvae originated from undergoing 1- and 44-generational cadmium treatment and in control (those that were not exposed to cadmium). We also measured the cadmium concentration and DNA damage level in the larvae. We observed higher level of heat shock proteins (HSPs) in the heads of larvae derived from multi-generational metal treatment than in the heads of those from one-generational treatment (derived from the control rearing). Analysis of HSP localisation in the larval brain suggests that these changes could be important for protecting the neural function of larval mushroom bodies for animals selected during multigenerational metal exposure. Animals from one-generational treatment had, in turn, higher total antioxidant capacity than animals from multigenerational treatment. Anyway, animals from one- and 44-generational metal treatments did not differ in metal accumulation in the heads and the whole larval bodies, catalase activity or DNA damage level. All these measurements were higher than for control larvae and cadmium accumulation in the heads was much lower than in the whole bodies.