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All processes involved in metal homeostasis must be coordinated to provide sufficient, but not toxic, concentrations of important bioelements, and to minimize detrimental effects of toxic metals. Our previous studies dealing with the exposure of O. nubilalis non-diapausing larvae to dietary Cd demonstrated that exposure to higher concentrations of Cd caused delay in the development of larvae, induced oxidative stress and also induced defense mechanisms against the toxic effects of Cd. The aim of the present study was to evaluate how acute and chronic exposure of O. nubilalis larvae to increased concentrations of dietary Cd affected the balance of important bioelements. The concentration of bioelements was analyzed in larvae (after short-term exposure) and pupae (after long-term exposure). The short-term exposure of final instar larvae (L5) to Cd did not affect significantly the concentration of any of the analyzed bioelements, while the long-term exposure of developing larvae to higher concentrations of Cd caused increase in the concentrations of Ca, Mg and Na in pupae. The bioaccumulation factor, calculated for bioelements after long-term exposure to Cd, was higher for the most bioelements in groups fed with diet containing higher concentrations of Cd, except K which displayed the opposite trend. Pearson correlation coefficient showed positive correlations between Cd and Ca, Mg, Na, Fe, Cu and Zn, while negative correlation was observed between Cd and K. The results indicate that impact on the balance of important bioelements might be one of the mechanisms of cadmium toxicity and certainly raise numerous questions for future research. © 2022, The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.

The use of insecticides, especially neonicotinoids (NEOCs), poses a significant threat to honey bees, thus prompting the EU to ban all outdoor uses of clothianidin, imidacloprid, and thiamethoxam. However, due to the persistence of NEOCs, as well as the fact that certain NEOCs are in use in some areas of the world, exposure to them is still an on-going issue. In the present study, we used laboratory acute toxicity test to examine the effects of sublethal concentrations (resulting in consumed doses approximately 50, 20, and 5 times below LD50) of thiacloprid and clothianidin on selected parameters of antioxidative defense (superoxide dismutase, catalase, glutathione S-transferase activities), oxidative status (reduced glutathione, protein thiols, malondialdehyde), neurotoxicity (acetylcholinesterase activity), and immune response (prophenoloxidase and phenoloxidase activities) in honey bees. Results indicated that both neonicotinoids’ sublethal concentrations promote oxidative stress in honey bees and affect their immune defense. NEOCs impair detoxification abilities through reduced glutathione S-transferase activity, as well as disturb immune response due to decreased prophenoloxidase and phenoloxidase activities, which can explain the high toxicity of these substances to honey bees. © 2022, INRAE, DIB and Springer-Verlag France SAS, part of Springer Nature.

The aim of the present study was to analyze the mechanisms of ibogaine action by measuring its ex vivo effects on antioxidant defense in the honey bee (Apis mellifera, L.) midgut. The transcriptional levels of selected genes: Cu/Zn dependent and Mn dependent superoxide dismutases (Sod1 and Sod2, respectively), catalase (Cat) and transcription factor Nrf2 (Nrf2) were determined. The applicability of midgut tissue, which expected to have well developed antioxidant protection system, for this type of analysis was confirmed by testing cell viability and response to paraquat, an effective inducer of oxidative stress, ex vivo. Incubation for 2 h with paraquat (10 µg/mL) induced a significant increase in expression of Sod1 and Cat genes. The results of ibogaine treatment showed that exposure to 5 µg/mL and 10 µg/mL of ibogaine for 2 h induced significant increase in expression of Sod1 gene. On the other hand, ibogaine did not lead to a significant increase of Sod2, Cat and transcription factor Nrf2 genes expression in honey bee midgut ex vivo. Our results confirmed positive effect of ibogaine on the antioxidant protective system and its pro-antioxidant action.

© 2019, Springer-Verlag GmbH Germany, part of Springer Nature. The enormous progress in nanomaterials development and their use, followed by their inevitable environmental print, has arisen the emerging questions concerning their influence to the living systems. Honey bees are considered to be quite a suitable model system for the risk assessment and prediction of various external influences. To the best of our knowledge, this is the first study dealing with the influence of fullerenol nanoparticles (FNP), a biodegradable carbon nanomaterials’ representative, to honey bees. This investigation was conducted with an aim to merge two different open-ended questions: the potential toxic effect of FNP to the bees on the one hand and antioxidative effect of FNP on the other hand. Since FNP antioxidative properties were proved in a number of in vivo models, we hypothesized the similar outcomes, and according to this assumption, we opted for paraquat as a well-known oxidative stress inducer. FNP did not have toxic effect in none of investigated concentrations. The results also confirmed the potential of FNP to reduce oxidative stress through the gene expression of antioxidative enzymes and the change in the redox state of the cells. Additional experiments are needed for a better understanding of the exact mechanism and complex patterns of FNP’s activity.

Cadmium (Cd) is a non-essential metal that is highly toxic to all living forms, characterized by an extremely high affinity for thiol (SH) groups. The aim of this work was to identify and experimentally verify metallothionein gene and to analyze the role of biological thiols in stress induced by short-term Cd exposure in Ostrinia nubilalis, one of the most important corn pests. The coding region of a metallothionein (MT) gene in O. nubilalis was identified, encoding protein, OnMT1, which contains 46 amino acids, including 12 cysteine residues, and has no aromatic amino acids. Phylogenetic analysis revealed that OnMT1 clustered together with metallothionein from Bombyx mori. Structural bioinformatics analysis strongly suggests that OnMT1 is a metallothionein with affinity for multiple transition metals. Further, in order to elucidate the role of biological thiols, O. nubilalis L5 larvae were exposed to increasing Cd concentrations in diet (6.85, 41.71, 77.35 mg kg-1) during a 48 h period, after which Cd concentration in larvae was measured (3.50, 12.02, 47.37 mg kg-1, respectively). Due to short-term Cd exposure, concentration of free protein SH groups and relative expression of OnMT1 and thioredoxin (Trx) genes was elevated, while the reduced glutathione content remained unchanged. The presented results provide evidence that OnMT1 plays a role in Cd detoxification and homeostasis, and confirm the importance of biological thiols, especially OnMT1 and Trx, in the early response of O. nubilalis to Cd poisoning, indicating interaction between Cd and thiol-linked redox reactions. Insects provide valuable insight into molecular adaptations to metals.

Honeybee colony losses have been a focus of research in the last years, due to the importance of managed honeybee colonies for economy and ecology. Different unfavorable conditions from the outside environment have a strong impact on the hive health. The majority of losses occur mainly during winter and the exact reason is not completely understood. Only a small number of studies are dealing with content of bioelements, their function and influence on honeybee physiology. The aim of the present study was to determine seasonal and spatial variations in content of bioelements and non-essential elements, in hemolymph and whole body of honeybees originating from three regions with different degrees of urbanization and industrialization. Concentrations of 16 elements were compared: macroelements (Ca, K, Mg, Na), microelements (Cu, Fe, Mn, Zn) and non-essential elements (Al, Ba, Cd, Co, Cr, Ni, Pb, Sr) in samples collected from 3 different environments: Golija (rural region), Belgrade (urban region) and Zajača (industrial region). Content of bioelements and non-essential elements in honeybees was under noticeable influence of the surrounding environment, season and degree of honeybee activity. Hemolymph was proven to be helpful in differentiating air pollution from other sources of honeybee exposure. The results of our study demonstrated that bees can be successfully used as biomonitors since we have observed statistically significant differences among observed locations, but unless compared locations are exposed to excessively different pollution pressures, it is essential that all bees should be collected at the same season. Supplementary material: [https://cherry.chem.bg.ac.rs/handle/123456789/4617] This is the peer-reviewed version of the article: Ilijević, K., Vujanović, D., Orčić, S. M., Purać, J., Kojić, D., Zarić, N. M., Gržetić, I., Blagojević, D. P.,& Čelić, T. V.. (2021). Anthropogenic influence on seasonal and spatial variation in bioelements and non-essential elements in honeybees and their hemolymph. in Comparative Biochemistry and Physiology Part - C: Toxicology and Pharmacology Elsevier Inc.., 239, 108852. [https://doi.org/10.1016/j.cbpc.2020.108852]

The European corn borer, Ostrinia nubilalis Hbn., is a pest Lepidopteran species whose larvae overwinter by entering diapause, gradually becoming cold-hardy. To investigate metabolic changes during cold hardening, activities of four metabolic enzymes – citrate synthase (CS), lactate dehydrogenase (LDH), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured in whole-body homogenates of pupae, non-diapausing and diapausing larvae acclimated to 5 °C, −3 °C and −16 °C. The highest CS activity was detected in non-diapausing larvae, reflecting active development, while the highest in vitro LDH activity was recorded in diapausing larvae at temperatures close to 0 °C, evidencing a metabolic switch towards anaerobic metabolism. However, in-gel LDH activity showed that production of pyruvate from lactate is triggered by sub-zero temperatures. The activities of both aminotransferases were highest in non-diapausing larvae. Our findings suggest that during diapause and cold hardening the aminotransferases catalyse production of L-alanine, an important cryoprotectant, and L-aspartate, which is closely tied to both transamination reactions and Krebs cycle. The results of this study indicate that, during diapause, the activity of metabolic enzymes is synchronized with exogenous factors, such as temperatures close to 0 °C. These findings support the notion that diapause is metabolically plastic and vibrant, rather than simply a passive, resting state.