Supplementary data for the article: Prokić, M. D.; Borković-Mitić, S. S.; Krizmanić, I. I.; Mutić, J. J.; Trifković, J. Đ.; Gavrić, J. P.; Despotović, S. G.; Gavrilović, B. R.; Radovanović, T. B.; Pavlović, S. Z.; et al. Bioaccumulation and Effects of Metals on Oxidative Stress and Neurotoxicity Parameters in the Frogs from the Pelophylax Esculentus Complex. Ecotoxicology 2016, 25 (8), 1531–1542. https://doi.org/10.1007/s10646-016-1707-x

Supplementary material for: [https://doi.org/10.1007/s10646-016-1707-x]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2340

Supplementary data for the article: Borković-Mitić, S. S.; Prokić, M. D.; Krizmanić, I. I.; Mutić, J.; Trifković, J.; Gavrić, J.; Despotović, S. G.; Gavrilović, B. R.; Radovanović, T. B.; Pavlović, S. Z.; et al. Biomarkers of Oxidative Stress and Metal Accumulation in Marsh Frog (Pelophylax Ridibundus). Environmental Science and Pollution Research 2016, 23 (10), 9649–9659. https://doi.org/10.1007/s11356-016-6194-3

Supplementary material for: [https://doi.org/10.1007/s11356-016-6194-3]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/1941

Supplementary data for article : Prokic, M. D.; Borkovic-Mitic, S. S.; Krizmanic, I. I.; Mutić, J.; Gavric, J. P.; Despotovic, S. G.; Gavrilović, B. R.; Radovanovic, T. B.; Pavlovic, S. Z.; Saicic, Z. S. Oxidative Stress Parameters in Two Pelophylax Esculentus Complex Frogs during Pre- and Post-Hibernation: Arousal vs Heavy Metals. Comparative Biochemistry and Physiology. C: Toxicology and Pharmacology 2017, 202, 19–25.https://doi.org/10.1016/j.cbpc.2017.07.00

Supplementary material for: [https://doi.org/10.1016/j.cbpc.2017.07.00]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2548

Supplementary data for the article: Borković-Mitić, S. S.; Prokić, M. D.; Krizmanić, I. I.; Mutić, J.; Trifković, J.; Gavrić, J.; Despotović, S. G.; Gavrilović, B. R.; Radovanović, T. B.; Pavlović, S. Z.; et al. Biomarkers of Oxidative Stress and Metal Accumulation in Marsh Frog (Pelophylax Ridibundus). Environmental Science and Pollution Research 2016, 23 (10), 9649–9659. https://doi.org/10.1007/s11356-016-6194-3

Supplementary material for: [https://doi.org/10.1007/s11356-016-6194-3]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/1941

Supplementary data for the article: Prokić, M. D.; Borković-Mitić, S. S.; Krizmanić, I. I.; Mutić, J. J.; Trifković, J. Đ.; Gavrić, J. P.; Despotović, S. G.; Gavrilović, B. R.; Radovanović, T. B.; Pavlović, S. Z.; et al. Bioaccumulation and Effects of Metals on Oxidative Stress and Neurotoxicity Parameters in the Frogs from the Pelophylax Esculentus Complex. Ecotoxicology 2016, 25 (8), 1531–1542. https://doi.org/10.1007/s10646-016-1707-x

Supplementary material for: [https://doi.org/10.1007/s10646-016-1707-x]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2340

Biochemical parameters in skin and muscle of Pelophylax kl. esculentus frogs: Influence of a cyanobacterial bloom in situ

There is little information in scientific literature as to how conditions created by a microcystin (MC) producing cyanobacterial bloom affect the oxidant/antioxidant, biotransformation and neurotoxicity parameters in adult frogs in situ. We investigated biochemical parameters in the skin and muscle of Pelophylax kl. esculentus from Lake Ludaš (Serbia) by comparing frogs that live on the northern bloom side (BS) of the lake with those that inhabit the southern no-bloom side (NBS). A higher protein carbonylation level and lower antioxidant defense system capability in the skin of frogs living in conditions of the cyanobacterial bloom were observed. Inhibition of glutathione-dependent machinery was the major mechanism responsible for the induction of cyanobacterial bloom-mediated oxidative stress in frog skin. On the other hand, the detected higher ability of muscle to overcome bloom prooxidant toxicity was linked to a higher efficiency of the biotransformation system through glutathione-S-transferase activity and/or was the consequence of indirect exposure of the tissue to the bloom. Our results have also revealed that the cyanobacterial bloom conditions induced the cholinergic neurotransmitter system in both tissues. This study provides a better understanding of the ecotoxicological impact of the MC producing cyanobacterial bloom on frogs in situ. However, further investigations of the complex mechanism involved in cyanobacterial bloom toxicity in real environmental conditions are required

Hepatic oxidative stress and neurotoxicity in Pelophylax kl. esculentus frogs: Influence of long-term exposure to a cyanobacterial bloom

Although the long-term exposure of aquatic organisms to cyanobacterial blooms is a regular occurrence in the environment, the prooxidant and neurotoxic effects of such conditions are still insufficiently investigated in situ. We examined the temporal dynamics of the biochemical parameters in the liver of Pelophylax kl. esculentus frogs that inhabit the northern (N) side of Lake Ludaš (Serbia) with microcystins (MCs) produced in a cyanobacterial bloom over three summer months. The obtained data were compared with data on frogs that live on the southern (S), MC-free side of the same lake. Our results showed that the MC-producing bloom induced oxidative damage to proteins and lipids, observed as a decrease in the concentration of protein -SH groups and increased lipid peroxidation (LPO) in the liver of N frogs in comparison to S frogs. Glutathione (GSH) played a key role in the transient defense against the MC-induced development of LPO. The low glutathione peroxidase (GPx) activity detected in all groups of frogs from the N site was crucial for the observed prooxidant consequences. The bloom impaired cholinergic homeostasis as a result of a decrease in ChE activity. A delayed neurotoxic effect in relation to the prooxidant outcomes was observed. Our results also showed that even though the integrated biomarker response (IBR) of the antioxidant biomarkers increased during exposure, the individual biochemical parameters did not exhibit a well-defined time-dependent pattern because of specific adaptation dynamics and/or additional effects of the physicochemical parameters of the water. This comprehensive environmental ecotoxicological evaluation of the cyanobacterial bloom-induced biochemical alterations in the liver of frogs provides a new basis for further investigations of the prolonged, real-life ecotoxicity of the blooms.The peer-reviewed version: [http://cer.ihtm.bg.ac.rs/handle/123456789/3677

Integrated response of antioxidant biomarkers in the liver and white muscle of European hake (Merluccius merluccius L.) females from the Adriatic sea with respect to environmental influences

We investigated the integrated response of antioxidant defense enzymes (total superoxide dismutase (TotSOD), manganese-containing superoxide dismutase (MnSOD), copper-zinc-containing superoxide dismutase (CuZnSOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione reductase (GR) and phase II biotransformation enzyme, glutathione- S-transferase (GST)) in the liver and white muscle of females of European hake (Merluccius merluccius L.) from the Adriatic Sea (Montenegro) in winter and spring. The activity of GSH-Px in the liver was significantly increased, while GST activity was decreased in spring compared to the winter. In white muscle, the activities of TotSOD and CuZnSOD were increased, while the activities of MnSOD, CAT, GSH-Px, GR and GST were decreased in spring when compared to the matching values in winter. The activities of TotSOD and CuZnSOD in winter were markedly lower in the muscle than in the liver, while the activity of MnSOD in the muscle was higher when compared to the liver. Principal component analysis (PCA) revealed clear separation of the investigated antioxidant biomarkers between tissues and seasons, while the integrated biomarker response (IBR) showed that the most intensive antioxidant biomarker response was in the liver in spring. Star plots of IBR showed a dominant contribution of glutathione-dependent biomarkers (GSH-Px, GR and GST) and CAT in both tissues and seasons with respect to SOD isoenzymes. All enzyme activities (except MnSOD) were greater in the liver in comparison to the white muscle. Our results show that the liver possesses a greater capacity to establish and maintain homeostasis under changing environmental conditions in winter and spring. At the same time, seasonal effects are more pronounced in muscle tissue. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. 173041

Oxidative Stress Parameters in Goitrogen-Exposed Crested Newt Larvae (Triturus spp.): Arrested Metamorphosis

Thiourea is an established disruptor of thyroid hormone synthesis and is frequently used as an inhibitor of metamorphosis. The changes caused by thiourea can affect processes associated with the oxidative status of individuals (metabolic rate, the HPI axis, antioxidant system). We investigated the parameters of oxidative stress in crested newt (Triturus spp.) larvae during normal development in late larval stage 62 and newly metamorphosed individuals, and during thiourea-stimulated metamorphosis arrest in individuals exposed to low (0.05%) and high (0.1%) concentrations of thiourea. Both groups of crested newts exposed to thiourea retained their larval characteristics until the end of the experiment. The low activities of antioxidant enzymes and the high lipid peroxidation level pointed to increased oxidative stress in larvae at the beginning of stage 62 as compared to fully metamorphosed individuals. The activities of catalase (CAT) and glutathione-S-transferase (GST) and the concentration of sulfhydryl (SH) groups were significantly lower in larvae reared in aqueous solutions containing thiourea than in newly metamorphosed individuals. The high thiourea concentration (0.1%) affected the antioxidative parameters to the extent that oxidative damage could not be avoided, contrary to a lower concentration. Our results provide a first insight into the physiological adaptations of crested newts during normal development and simulated metamorphosis arrest

Effects of metals on blood oxidative stress biomarkers and acetylcholinesterase activity in dice snakes (Natrix tessellata) from Serbia

The effects of waterborne metals in water on the activities of blood copper-zinc superoxide dismutase (CuZnSOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione reductase (GR), glutathione-S-transferase (GST), and acetylcholinesterase (AChE), and on the concentrations of total glutathione (GSH) and lipid peroxides (TBARS) in the blood of dice snakes (Natrix tessellata) caught in Obedska Bara, Sebia (control area), with snakes caught in Pančevački Rit, a contaminated area in Serbia were examined. The activities of CAT, GSH-Px, GR and AChE, and the concentration of TBARS were significantly decreased, while GST activity and GSH concentration were significantly increased in snakes from the contaminated area compared to specimens from the control area. Significantly increased concentrations of Al, As, B, Ba, Ca, Cu, Fe, K, Li, Mn, Na, Ni and Zn in the water at the contaminated area as compared to control area were detected. The metals Ag, Bi, Cd, Co, Hg, In and Tl were not observed in any of the localities. Cr, Mo and Pb were not detected at the control area but were observed at the contaminated area. The concentrations of Sr were similar at both sites. The concentration of Mg was 2-fold higher at the control site than at the contaminated area. The obtained results show that most of the investigated blood biomarkers correlate with concentrations of metals present in the environment. These findings suggest that dice snakes are sensitive bioindicator species for monitoring the effects of increased metal concentrations in the environment. [Projekat Ministarstva nauke Republike Srbije, br. 173041 i br. 173043