Concentration- and Time-Dependent Dietary Exposure to Graphene Oxide and Silver Nanoparticles: Effects on Food Consumption and Assimilation, Digestive Enzyme Activities, and Body Mass in Acheta domesticus.

The advancement of nanotechnology poses a real risk of insect exposure to nanoparticles (NPs) that can enter the digestive system through contaminated food or nanopesticides. This study examines whether the exposure of model insect species-Acheta domesticus-to increasing graphene oxide (GO) and silver nanoparticle (AgNP) concentrations (2, 20, and 200 ppm and 4, 40, and 400 ppm, respectively) could change its digestive functions: enzymes' activities, food consumption, and assimilation. We noticed more pronounced alterations following exposure to AgNPs than to GO. They included increased activity of α-amylase, α-glucosidase, and lipase but inhibited protease activity. Prolonged exposure to higher concentrations of AgNPs resulted in a significantly decreased food consumption and changed assimilation compared with the control in adult crickets. A increase in body weight was observed in the insects from the Ag4 group and a decrease in body weight or no effects were observed in crickets from the Ag40 and Ag400 groups (i.e., 4, 40, or 400 ppm of AgNPs, respectively), suggesting that even a moderate disturbance in nutrient and energy availability may affect the body weight of an organism and its overall condition. This study underscores the intricate interplay between NPs and digestive enzymes, emphasizing the need for further investigation to comprehend the underlying mechanisms and consequences of these interactions.

Survival, growth and digestive functions after exposure to nanodiamonds - Transgenerational effects beyond contact time in house cricket strains.

The long-term exposure effects of nanodiamonds (NDs), spanning an organism's entire lifespan and continuing for subsequent generation, remain understudied. Most research has focused on evaluating their biological impacts on cell lines and selected organisms, typically over short exposure durations lasting hours or days. The study aimed to assess growth, mortality, and digestive functions in wild (H) and long-lived (D) strains of Acheta domesticus (Insecta: Orthoptera) after two-generational exposure to NDs in concentrations of 0.2 or 2 mg kg-1 of food, followed by their elimination in the third generation. NDs induced subtle stimulating effect that depended on the strain and generation. In the first generation, more such responses occurred in the H than in the D strain. In the first generation of H strain insects, contact with NDs increased survival, stimulated the growth of young larvae, and the activity of most digestive enzymes in mature adults. The same doses and exposure time did not cause similar effects in the D strain. In the first generation of D strain insects, survival and growth were unaffected by NDs, whereas, in the second generation, significant stimulation of those parameters was visible. Selection towards longevity appears to support higher resistance of the insects to exposure to additional stressor, at least in the first generation. The cessation of ND exposure in the third generation caused potentially harmful changes, which included, e.g., decreased survival probability in H strain insects, slowed growth of both strains, as well as changes in heterochromatin density and distribution in nuclei of the gut cells in both strains. Such a reaction may suggest the involvement of epigenetic inheritance mechanisms, which may become inadequate after the stress factor is removed.

Multigenerational Effects of Graphene Oxide Nanoparticles on Acheta domesticus DNA Stability.

The use of nanoparticles like graphene oxide (GO) in nanocomposite industries is growing very fast. There is a strong concern that GO can enter the environment and become nanopollutatnt. Environmental pollutants' exposure usually relates to low concentrations but may last for a long time and impact following generations. Attention should be paid to the effects of nanoparticles, especially on the DNA stability passed on to the offspring. We investigated the multigenerational effects on two strains (wild and long-lived) of house cricket intoxicated with low GO concentrations over five generations, followed by one recovery generation. Our investigation focused on oxidative stress parameters, specifically AP sites (apurinic/apyrimidinic sites) and 8-OHdG (8-hydroxy-2'-deoxyguanosine), and examined the global DNA methylation pattern. Five intoxicated generations were able to overcome the oxidative stress, showing that relatively low doses of GO have a moderate effect on the house cricket (8-OHdG and AP sites). The last recovery generation that experienced a transition from contaminated to uncontaminated food presented greater DNA damage. The pattern of DNA methylation was comparable in every generation, suggesting that other epigenetic mechanisms might be involved.

Does age pay off? Effects of three-generational experiments of nanodiamond exposure and withdrawal in wild and longevity-selected model animals.

Nanodiamonds (NDs) are considered a material with low toxicity. However, no studies describe the effects of ND withdrawal after multigenerational exposure. The aim was to evaluate ND exposure (in the 1st and 2nd generations) effects at low concentrations (0.2 or 2 mg kg-1) and withdrawal (in the 3rd generation) in the wild (H) and longevity-selected (D) model insect Acheta domesticus. We measured selected oxidative stress parameters, immunity, types of cell death, and DNA damage. Most of the results obtained in the 1st generation, e.g., catalase (CAT), total antioxidant capacity (TAC), heat shock proteins (HSP70), defensins, or apoptosis level, confirmed no significant toxicity of low doses of NDs. Interestingly, strain-specific differences were observed. D-strain crickets reduced autophagy, the number of ROS+ cells, and DNA damage. The effect can be a symptom of mobilization of the organism and stimulation of physiological defense mechanisms in long-living organisms. The 2nd-generation D-strain insects fed ND-spiked food at higher concentrations manifested a reduction in CAT, TAC, early apoptosis, and DNA damage, together with an increase in HSP70 and defensins. ROS+ cells and cells with reduced membrane potential and autophagy did not differ significantly from the control. H-strain insects revealed a higher number of ROS+ cells and cells with reduced membrane potential, decreased CAT activity, and early apoptosis. Elimination of NDs from the diet in the 3rd generation did not cause full recovery of the measured parameters. We noticed an increase in the concentration of HSP70 and defensins (H-strain) and a decrease in apoptosis (D-strain). However, the most visible increase was a significant increase in DNA damage, especially in H-strain individuals. The results suggest prolonged adverse effects of NDs on cellular functions, reaching beyond "contact time" with these particles. Unintentional and/or uncontrolled ND pollution of the environment poses a new challenge for all organisms inhabiting it, particularly during multigenerational exposure.

Age- and Lifespan-Dependent Differences in GO Caused DNA Damage in Acheta domesticus.

The rising applicability of graphene oxide (GO) should be preceded by detailed tests confirming its safety and lack of toxicity. Sensitivity to GO of immature, or with different survival strategy, individuals has not been studied so far. Therefore, in the present research, we focused on the GO genotoxic effects, examining selected parameters of DNA damage (total DNA damage, double-strand breaks-DSB, 8-hydroxy-2'-deoxyguanosine-8-OHdG, abasic site-AP sites), DNA damage response parameters, and global methylation in the model organism Acheta domesticus. Special attention was paid to various life stages and lifespans, using wild (H), and selected for longevity (D) strains. DNA damage was significantly affected by stage and/or strain and GO exposure. Larvae and young imago were generally more sensitive than adults, revealing more severe DNA damage. Especially in the earlier life stages, the D strain reacted more intensely/inversely than the H strain. In contrast, DNA damage response parameters were not significantly related to stage and/or strain and GO exposure. Stage-dependent DNA damage, especially DSB and 8-OHdG, with the simultaneous lack or subtle activation of DNA damage response parameters, may result from the general life strategy of insects. Predominantly fast-living and fast-breeding organisms can minimize energy-demanding repair mechanisms.

Multigenerational selection towards longevity changes the protective role of vitamin C against graphene oxide-induced oxidative stress in house crickets.

This research was designed to investigate changes that can arise in an invertebrate organism due to stress caused by a strong prooxidant, graphene oxide (GO), and a potent antioxidant, vitamin C. The study aimed to investigate if vitamin C may support convalescence after chronic GO intoxication. We investigated the toxicity of chronic dietary graphene oxide administration in house cricket (Acheta domesticus) types: wild and selected for longevity (with a better developed antioxidant system, conducive to long life). Vitamin C was applied immediately after cessation of graphene oxide intoxication to check if it can support the remedial effect. The condition of cells, DNA stability, catalase activity, and the reproduction potential, measured as the Vitellogenin (Vg) protein expression level, were investigated in control and GO treated groups, recovery groups (-GO), and recovery groups with Vit. C (-GO + Vit.C). In this study vitamin C had no evident remedial effect on the house crickets exposed to graphene oxide. Most probably, the mechanism of vitamin C action, in case of intoxication with nanoparticles, is much more complicated. In the context of the results obtained, it is worth considering whether Vit. C, applied after GO intoxication, causes further disturbance of homeostasis in terms of the cells' redox potential.

Multigenerational graphene oxide intoxication results in reproduction disorders at the molecular level of vitellogenin protein expression in Acheta domesticus.

The anthropogenic activities may lead to accumulation of graphene oxide (GO) pollution in the environment. Organisms exposed to chronic or multigenerational GO intoxication can present reproduction depletion. Vitellogenin (Vg) has been used as a parameter for evaluating female fertility due to its importance in embryo nutrition. In this study, we used a promising model organism, Acheta domesticus, which was intoxicated with GO in food for three generations. The aim of the study was to investigate the process of Vg synthesis in crickets depending on the exposure time, GO concentration, and age of the females. The results revealed that chronic GO intoxication had adverse effects on the Vg expression pattern. The 1st generation of insects showing low Vg expression was most affected. The 2nd generation of A. domesticus presented a high Vg expression. The last investigated generation seemed to cope with stress caused by GO, and the Vg expression was balanced. We suggest that the epigenetic mechanisms may play a role in the information transfer to the next generations on how to react to the risk factor and keep reproduction at a high rate. We suspect that chronic GO intoxication can disturb the regular formation of the Vg quaternary structure, resulting in consequences for developing an embryo.

Do nanoparticles cause hormesis? Early physiological compensatory response in house crickets to a dietary admixture of GO, Ag, and GOAg composite.

This study aimed to identify the physiological responses of house cricket females following short-term exposure to relatively low dietary doses of graphene oxide (GO, 20 µg · g-1 food), silver (Ag, 400 µg · g-1 food) nanoparticles (NPs), or graphene oxide­silver nanoparticle composite (GO-AgNPs, 20: 400 µg · g-1 food). Energy intake and distribution were measured on the third, sixth, and tenth day. A semi-quantitative API®ZYM assay of digestive enzyme fingerprints was performed on the third and tenth day of continuous treatment. Physicochemical properties of the NPs were obtained by combining SEM, EDX spectrometry, AFM, and DLS techniques. The obtained results showed decreased energy consumption, particularly assimilation as an early response to dietary NPs followed by compensatory changes in feeding activity leading to the same consumption and assimilation throughout the experimental period (10 days). The increased activities of digestive enzymes in NP-treated females compared to the control on the third day of the experiment suggest the onset of compensatory reactions of the day. Moreover, the insects treated with GO-AgNP composite retained more body water, suggesting increased uptake. The observed changes in the measured physiological parameters after exposure to NPs are discussed in light of hormesis.

Towards understanding Cameraria ohridella (Lepidoptera: Gracillariidae) development: effects of microhabitat variability in naturally growing horse-chestnut tree canopy.

Dwelling intensity of horse-chestnut miner (Cameraria ohridella) larvae in various leaves insolation and temperature was measured to determine whether this pest's development follows a predictable pattern or depends more on local microenvironment conditions. Mines growing on leaves of mature host plants (Aesculus hippocastanum L.) in their natural conditions were photographed for two consecutive generations of the pest and in two separated vegetation periods. Apart from meteorological data obtained from the nearest station, the temperature of intact and mined parts of sun-exposed and shaded leaf blades was measured at various daytimes throughout the experiment. Obtained sets of digital data were analysed and combined to model mine area growth as a function of degree-days sum by adopting of Verhulst logistic equation. We showed the predictive potential of our model based on experimental data, and it may be useful in the scheduling of pest control measures in natural conditions. Our analyses also revealed that despite significant differences in microenvironment conditions depending on mines' insolation, the horse-chestnut miner larvae could partially compensate for them and complete their development at similar endpoints expressed as the cumulative sum of degree-days. We conclude that computer-aided analysis of photographic documentation of leaf-miner larval growth followed by mathematical modelling offers a noninvasive, reliable, and inexpensive alternative for monitoring local leaf-miners populations.

Vitellogenin expression, DNA damage, health status of cells and catalase activity in Acheta domesticus selected according to their longevity after graphene oxide treatment.

The increased use of graphene oxide (GO) raises worrisome questions regarding its possible threat to various ecosystems. Invertebrates represent valuable organisms for environmental studies. The lifespan can influence the ability to cope with toxins, especially those that act via oxidative stress. Two strains of Acheta domesticus, which are selected for longevity, were tested. The main aim was to investigate how GO, when administrated in food, affects: the condition of cells, DNA stability, ROS generation and the reproduction potential (the Vitellogenin (Vg) protein expression). The "recovery effect" - after removing GO from the diet for 15 days - was also measured. The results revealed different responses to GO in the wild (H) and long-living (D) strains. The D strain had a higher catalase activity compared to the H strain on the 25th day of the imago stage. Removing GO from the food resulted in a decrease in the catalase activity to the level of the control. On the 5th day of the imago stage, the H strain had a higher cell mortality than the D strain in the GO-intoxicated groups. There was more DNA damage in the H strain compared to the long-living strain. A remedial effect was seen after the GO was removed from the diet. The total Vg protein expression was higher in the H strain and lower in the D strain. The results indicated a GO concentration-dependent outcome. In both strains, removing the GO from the food led to a high Vg expression. The Vg expression after GO treatment, particularly translation and post-translational processing, should be studied in detail in the future. The D strain of crickets had more specialized mechanisms for maintaining homeostasis than the H strain. Organisms can fight off negative effects of GO, especially when they have systems that are well developed against oxidative stress.

Graphene oxide as a new anthropogenic stress factor - multigenerational study at the molecular, cellular, individual and population level of Acheta domesticus.

Although interest in transgenerational phenomena is constantly growing, little is known about the long-term toxicity of nanoparticles. In this study we investigate the multigenerational effects of graphene oxide (GO) which was given to Acheta domesticus in low doses (0.2, 2 and 20 µg·g-1 of food) for three subsequent generations. We assessed the influence of GO nanoparticles in many contexts, basing on parameters which represented different levels of biological organization: activity of antioxidant enzymes, level of apoptosis, DNA damage, histological analysis, hatching abilities, body mass and body length of insects, as well as their survival rate. The results have shown that exposing insects to nanoparticles over an extended period of time causes surprising intergenerational effects, based on significant differences in the life cycle and reproductive processes, which are not always dose-dependent. The second generation of insects appeared as the most unstable among the parameters that were studied, and did not match trends and patterns in the first and third generation categories. An increase of DNA damage was observed, but only in the third generation. This reduction of genome stability can be perceived as an essential element of adaptation, leading to an increase of genotype variants, which then undergo selection.

Enhancement of and interference among higher order multipole transitions in molecules near a plasmonic nanoantenna.

Spontaneous emission of quantum emitters can be modified by their optical environment, such as a resonant nanoantenna. This impact is usually evaluated under assumption that each molecular transition is dominated only by one multipolar channel, commonly the electric dipole. In this article, we go beyond the electric dipole approximation and take light-matter coupling through higher-order multipoles into account. We investigate a strong enhancement of the magnetic dipole and electric quadrupole emission channels of a molecule adjacent to a plasmonic nanoantenna. Additionally, we introduce a framework to study interference effects between various transition channels in molecules by rigorous quantum-chemical calculations of their multipolar moments and a consecutive investigation of the transition rate upon coupling to a nanoantenna. We predict interference effects between these transition channels, which allow in principle for a full suppression of radiation by exploiting destructive interference, waiving limitations imposed on the emitter's coherence time by spontaneous emission.

Evaluation of selected biological properties of the hunting web spider (Steatoda grossa, Theridiidae) in the aspect of short- and long-term exposure to cadmium.

The study aimed at comparing the effects of short- and long-term exposure of Steatoda grossa female spiders to cadmium on the web's architecture, its energy content, and ultrastructure of ampullate glands. Simple food chain model (medium with 0.25 mM CdCl2 → Drosophila hydei flies → spider (for 4 weeks or 12 months) was used for the exposure. Analysis of Cd content provided evidence that silk fibers of the web are well protected against its incorporation irrespectively of the exposure period. Long-term exposure to cadmium resulted in the occurrence of numerous autophagosomes with degenerated organelles as well as apoptotic and necrotic cells in the ampullate glands. Concurrently, the individual silk fibers building double and multiple combination complexes were significantly thinner than in the control threads. Moreover, exposed spiders spun net with smaller mean calorific value than did the control individuals. Hence, evaluation of both the diameter of silk fibers and calorific value of the web can serve as biomarkers of the effects caused by exposure of these predators to cadmium.

Chronic toxicity of nanodiamonds can disturb development and reproduction of Acheta domesticus L.

The use of nanodiamonds in numerous materials designed for industry and medicine is growing rapidly. Consequently health and environmental risks associated with the exposure of humans and other biota to nanodiamonds-based materials are of the utmost importance. Scarcity of toxicological data for these particles led us to examine the potentially deleterious effects of nanodiamonds in model insect species, Acheta domesticus (Orthoptera) chronically exposed to ND in its diet. Organism-level end-point indices (lifespan, body weight, consumption, caloric value of faeces, reproduction) revealed adverse changes in the treated crickets in comparison with the control. Preliminary studies of oxidative stress level in the offspring of ND-treated crickets suggest toxicity of these particles limited to the exposed individuals. EPR analysis showing increase of radical signal in the faeces of ND-fed crickets led us to propose novel mechanism of nanodiamonds toxicity that is discussed in the light of literature data. CAPSULE: Development and reproduction of Acheta domesticus can be disturbed by the chronic exposure to nanodiamonds.

Lifespan differences between queens and workers are not explained by rates of molecular damage.

The biological processes that underlie senescence are of universal biological importance, yet they remain poorly understood. A popular theory proposes that senescence is the result of limited investment into mechanisms involved in the prevention and repair of molecular damage, leading to an accumulation of molecular damage with age. In ants, queen and worker lifespans differ by an order of magnitude, and this remarkable difference in lifespan has been shown to be associated with differences in the expression of genes involved in DNA and protein repair. Here we use the comet assay and Western Blotting for poly-ubiquitinated proteins to explore whether these differences in expression lead to differences in the accumulation of DNA damage (comet assay) or protein damage (protein ubiquitination) with age. Surprisingly, there was no difference between queens and workers in the rate of accumulation of DNA damage. We also found that levels of ubiquitinated proteins decreased with age, as previously reported in honeybees. This is in contrast to what has been found in model organisms such as worms and flies. Overall, these results reveal that the link between investment into macromolecular repair, age-related damage accumulation and lifespan is more complex than usually recognised.

The effect of ingested cadmium on the calorific value and structural properties of hunting webs produced by Steatoda grossa (Theridiidae) spiders.

The study aimed to assess whether cadmium administered via ingestion to Steatoda grossa cobweb spiders (Theridiidae) affects the energy content and selected structural properties of the produced hunting webs. Cadmium content in webs was assessed with AAS and SEM X-ray microanalysis, while the diameters of silk fibers were estimated with SEM. The energy content of samples was measured in an oxygen micro-bomb calorimeter. Females and males showed different reactions to cadmium supplied through food. In comparison to females, males displayed higher metal concentrations in their bodies and hunting webs, however their calorific values and structural features were not significantly changed. Cadmium-treated females spun webs with smaller single-strand diameters and more frequent multi-stranded threads and invested 47% less energy in web production than the control individuals. It cannot be excluded that such a reduction in energy expenditure for web building in females resulted from energetically costly detoxifying reactions triggered in response to direct and indirect effects of cadmium toxicity.

The first example of ab initio calculations of f-f transitions for the case of [Eu(DOTP)]5- complex-experiment versus theory.

Crystal structures and photophysical properties (IR and UV-vis-NIR) of two compounds, [C(NH2)3]5[Eu(DOTP)]·12.5H2O and K5[Eu(DOTP)]·11H2O (DOTP = 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrakis (methylenephosphonic acid)), were determined. The DOTP ligand is bonded to Eu3+via four O and four N atoms, filling thus eight coordination sites of Eu3+. The experimental structures of two [K4Eu(DOTP)]- clusters were used as a starting point for theoretical ab initio calculations based on a multireference wavefunction approach. Positions of the energy levels of the 4f6 configuration of the Eu3+ ion have been calculated and compared with those derived from the experimental spectra. This enabled us to tentatively assign energy levels of the Eu3+ ion. The relationship between calculated energies of excited states and Eu-N and Eu-O bond lengths was discussed with respect to the nephelauxetic effect.

Parnassius apollo last-instar larvae development prediction by analysis of weather condition as a tool in the species' conservation.

Parnassius apollo (Lepidoptera: Papilionidae) has already disappeared or is under threat of extinction in many of its former habitats. It has been documented that weather conditions--anomalies in particular--contributed to this process. In this study, we combined developmental data obtained previously for the last-instar Apollo larvae (collected in 1996, 1997, and 2003) with corresponding meteorological data to assess the effects of ambient temperature and rainfall episodes on the duration and the completion of the instar. For comparing the temperature effect, we applied the degree-day concept. We found significant positive correlation between the number of rainy days during the instar development (x) and its duration time (y): y=8.293+0.936x (±2.813) (r=0.662, P<10(-7)). Logarithmic transformation of the growth curves of the last-instar Apollo larvae revealed that there was no difference in growth among females; however, there was slower growth of males in 2003 in comparison to 1996. Growth (y) of female Apollo larvae as a function of instar duration (x) can by described by one common equation, irrespectively of the year: y=317.6+502.3 lnx (±263.3) (r=0.82, P<10(-4)).