TY  - CONF
AU  - Grčić, Anja
AU  - Ilijin, Larisa
AU  - Filipović, Aleksandra
AU  - Matić, Dragana
AU  - Mrdaković, Marija
AU  - Todorović, Dajana
AU  - Perić Mataruga, Vesna
PY  - 2021
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/4913
AB  - INTRODUCTION: Intense anthropogenic influence led to a significant increase in pollution of the biosphere, in which polycyclic aromatic hydrocarbons, especially benzo[a]pyrene (B[a]P) made a major contribution. Wet and dry deposition gets atmospheric B[a]P on the vegetation, an important sink, and a crucial link for B[a]P bioaccumulation in animals1. The gypsy moth is a phytophagous polyphagous insect that inhabits wide forest areas. Due to its vast appetite, it can pile great amounts of pollutants making it a suitable model system for biomonitoring the adverse effects of B[a]P. The larval midgut is the central metabolic place where trypsin and lipases provide efficient digestion of protein and lipids-rich food, showing sensitivity to chemical pollutants2,3. Molecular parameters can be affected by physiological and environmental factors, so different adaptations of insects to the contaminants should be considered during the assessment of biomarker potential. OBJECTIVES: The aim was to investigate chronic effects of dietary treatment with B[a]P on midgut enzyme activities of trypsin and lipase, as well as the content of total proteins and lipids in hemolymph in gypsy moth larvae from two populations - one from an unpolluted oak forest and the other from a polluted oak forest. Furthermore, we used the method of Integrated Biomarker Response (IBR) to summarize responses of multiple molecular parameters across different tissues to estimate their sensitivity to B[a]P exposure in terms of population origin. METHOD / DESIGN: Gypsy moth egg masses were gathered in two mixed oak forests – Đerdap National Park forest, free of industrial pollution (unpolluted population of larvae, UP), and Bor forest contaminated by various byproducts of the mining industry (polluted population of larvae, PP). From hatching until the sacrifice (third day of the 5th instar) larvae were fed with a diet containing 0 ng (UP 0 ng and PP 0 ng), 5 ng (UP 5 ng and PP 5 ng), or 50 ng (UP 50 ng and PP 50 ng) of B[a]P in 1 g of dry diet. Spectrophotometric assays were used for the determination of specific enzyme activities of trypsin and lipase in the homogenates of the midgut, as well as for the evaluation of total proteins and lipids in the hemolymph of larvae4-8. Two-way ANOVA followed by Tukey’s post-hoc test was used for statistical analyses, conducted in GraphPad Prism 8 (GraphPad Software, Inc., USA). Statistical significance was determined at probability (p)<0.05. Excel software (Microsoft, USA) was used to calculate IBR values and to generate star plots9. RESULTS: The specific activity of trypsin has significantly inhibited after the treatment with lower B[a]P concentration in UP (F=9.412, p=0.0004), while a higher concentration of B[a]P significantly induced lipase activity in the same population of larvae (F=8.382, p=0.0007). These enzymes showed no statistically significant changes in the PP. Hemolymph protein content was significantly affected by the chronic dietary exposure to the higher concentration of B[a]P in both populations of larvae, showing a decrease in the UP, and the elevation in the PP (F=10.16, p=0.0002). Lipid concentration was not significantly changed under the B[a]P influence regarding the control groups (UP/PP 0 ng) but there was a meaningful difference between B[a]P treated larvae among two populations (F=7.16, p=0.019). IBR index increased in a concentration-dependent way only in UP after the chronic exposure to B[a]P and the values were higher than the corresponding ones in the PP (IBR index values - UP 0 ng=0; UP 5 ng=1.62; UP 50 ng=4.84; PP 0 ng=2.01; PP 5 ng=1.10; PP 50 ng=3.08). CONCLUSIONS: Gypsy moth population from the unpolluted forest showed higher sensitivity to the chronic dietary exposure to B[a]P comparing to the population from the polluted forest, especially in terms of trypsin and lipase activity. Hemolymph protein level expressed well correspondence to B[a]P concentration in both populations but with inverse trends. The selected set of Gypsy moth larvae molecular parameters possess a good potential for B[a]P biomonitoring in the populations from unpolluted forest ecosystems.
PB  - Novi Sad: Faculty of Sciences, University of Novi Sad
C3  - Book of abstracts: The International Bioscience Conference and the 8th International PSU - UNS Bioscience Conference IBSC 2021; 2021 Nov 25-26; Novi Sad, Serbia
T1  - Midgut trypsin and lipase activities, hemolymph protein and lipids levels with integrated biomarker response (Ibr) in Gypsy moth (Lymantria Dispar) larvae from clean and polluted forest after chronic exposure to benzo[a]pyrene
SP  - 65
EP  - 66
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_4913
ER  - 

@conference{
editor = "Mimica-Dukić, Neda, Pajević, Slobodanka, Mandić, Anamarija",
author = "Grčić, Anja and Ilijin, Larisa and Filipović, Aleksandra and Matić, Dragana and Mrdaković, Marija and Todorović, Dajana and Perić Mataruga, Vesna",
year = "2021",
abstract = "INTRODUCTION: Intense anthropogenic influence led to a significant increase in pollution of the biosphere, in which polycyclic aromatic hydrocarbons, especially benzo[a]pyrene (B[a]P) made a major contribution. Wet and dry deposition gets atmospheric B[a]P on the vegetation, an important sink, and a crucial link for B[a]P bioaccumulation in animals1. The gypsy moth is a phytophagous polyphagous insect that inhabits wide forest areas. Due to its vast appetite, it can pile great amounts of pollutants making it a suitable model system for biomonitoring the adverse effects of B[a]P. The larval midgut is the central metabolic place where trypsin and lipases provide efficient digestion of protein and lipids-rich food, showing sensitivity to chemical pollutants2,3. Molecular parameters can be affected by physiological and environmental factors, so different adaptations of insects to the contaminants should be considered during the assessment of biomarker potential. OBJECTIVES: The aim was to investigate chronic effects of dietary treatment with B[a]P on midgut enzyme activities of trypsin and lipase, as well as the content of total proteins and lipids in hemolymph in gypsy moth larvae from two populations - one from an unpolluted oak forest and the other from a polluted oak forest. Furthermore, we used the method of Integrated Biomarker Response (IBR) to summarize responses of multiple molecular parameters across different tissues to estimate their sensitivity to B[a]P exposure in terms of population origin. METHOD / DESIGN: Gypsy moth egg masses were gathered in two mixed oak forests – Đerdap National Park forest, free of industrial pollution (unpolluted population of larvae, UP), and Bor forest contaminated by various byproducts of the mining industry (polluted population of larvae, PP). From hatching until the sacrifice (third day of the 5th instar) larvae were fed with a diet containing 0 ng (UP 0 ng and PP 0 ng), 5 ng (UP 5 ng and PP 5 ng), or 50 ng (UP 50 ng and PP 50 ng) of B[a]P in 1 g of dry diet. Spectrophotometric assays were used for the determination of specific enzyme activities of trypsin and lipase in the homogenates of the midgut, as well as for the evaluation of total proteins and lipids in the hemolymph of larvae4-8. Two-way ANOVA followed by Tukey’s post-hoc test was used for statistical analyses, conducted in GraphPad Prism 8 (GraphPad Software, Inc., USA). Statistical significance was determined at probability (p)<0.05. Excel software (Microsoft, USA) was used to calculate IBR values and to generate star plots9. RESULTS: The specific activity of trypsin has significantly inhibited after the treatment with lower B[a]P concentration in UP (F=9.412, p=0.0004), while a higher concentration of B[a]P significantly induced lipase activity in the same population of larvae (F=8.382, p=0.0007). These enzymes showed no statistically significant changes in the PP. Hemolymph protein content was significantly affected by the chronic dietary exposure to the higher concentration of B[a]P in both populations of larvae, showing a decrease in the UP, and the elevation in the PP (F=10.16, p=0.0002). Lipid concentration was not significantly changed under the B[a]P influence regarding the control groups (UP/PP 0 ng) but there was a meaningful difference between B[a]P treated larvae among two populations (F=7.16, p=0.019). IBR index increased in a concentration-dependent way only in UP after the chronic exposure to B[a]P and the values were higher than the corresponding ones in the PP (IBR index values - UP 0 ng=0; UP 5 ng=1.62; UP 50 ng=4.84; PP 0 ng=2.01; PP 5 ng=1.10; PP 50 ng=3.08). CONCLUSIONS: Gypsy moth population from the unpolluted forest showed higher sensitivity to the chronic dietary exposure to B[a]P comparing to the population from the polluted forest, especially in terms of trypsin and lipase activity. Hemolymph protein level expressed well correspondence to B[a]P concentration in both populations but with inverse trends. The selected set of Gypsy moth larvae molecular parameters possess a good potential for B[a]P biomonitoring in the populations from unpolluted forest ecosystems.",
publisher = "Novi Sad: Faculty of Sciences, University of Novi Sad",
journal = "Book of abstracts: The International Bioscience Conference and the 8th International PSU - UNS Bioscience Conference IBSC 2021; 2021 Nov 25-26; Novi Sad, Serbia",
title = "Midgut trypsin and lipase activities, hemolymph protein and lipids levels with integrated biomarker response (Ibr) in Gypsy moth (Lymantria Dispar) larvae from clean and polluted forest after chronic exposure to benzo[a]pyrene",
pages = "65-66",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_4913"
}

Mimica-Dukić, N., Pajević, S., Mandić, A., Grčić, A., Ilijin, L., Filipović, A., Matić, D., Mrdaković, M., Todorović, D.,& Perić Mataruga, V.. (2021). Midgut trypsin and lipase activities, hemolymph protein and lipids levels with integrated biomarker response (Ibr) in Gypsy moth (Lymantria Dispar) larvae from clean and polluted forest after chronic exposure to benzo[a]pyrene. in Book of abstracts: The International Bioscience Conference and the 8th International PSU - UNS Bioscience Conference IBSC 2021; 2021 Nov 25-26; Novi Sad, Serbia
Novi Sad: Faculty of Sciences, University of Novi Sad., 65-66.
https://hdl.handle.net/21.15107/rcub_ibiss_4913

Mimica-Dukić N, Pajević S, Mandić A, Grčić A, Ilijin L, Filipović A, Matić D, Mrdaković M, Todorović D, Perić Mataruga V. Midgut trypsin and lipase activities, hemolymph protein and lipids levels with integrated biomarker response (Ibr) in Gypsy moth (Lymantria Dispar) larvae from clean and polluted forest after chronic exposure to benzo[a]pyrene. in Book of abstracts: The International Bioscience Conference and the 8th International PSU - UNS Bioscience Conference IBSC 2021; 2021 Nov 25-26; Novi Sad, Serbia. 2021;:65-66.
https://hdl.handle.net/21.15107/rcub_ibiss_4913 .

Mimica-Dukić, Neda, Pajević, Slobodanka, Mandić, Anamarija, Grčić, Anja, Ilijin, Larisa, Filipović, Aleksandra, Matić, Dragana, Mrdaković, Marija, Todorović, Dajana, Perić Mataruga, Vesna, "Midgut trypsin and lipase activities, hemolymph protein and lipids levels with integrated biomarker response (Ibr) in Gypsy moth (Lymantria Dispar) larvae from clean and polluted forest after chronic exposure to benzo[a]pyrene" in Book of abstracts: The International Bioscience Conference and the 8th International PSU - UNS Bioscience Conference IBSC 2021; 2021 Nov 25-26; Novi Sad, Serbia (2021):65-66,
https://hdl.handle.net/21.15107/rcub_ibiss_4913 .

TY  - CONF
AU  - Matić, Dragana
AU  - Vlahović, Milena
AU  - Ilijin, Larisa
AU  - Grčić, Anja
AU  - Filipović, Aleksandra
AU  - Mrdaković, Marija
AU  - Perić Mataruga, Vesna
PY  - 2021
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/4912
AB  - INTRODUCTION:
Cadmium (Cd) levels in the environment have increased during decades of intensive industrial development and urbanization. Lymantria dispar has proved to be a suitable organism indicator to monitor Cd pollution in the forest ecosystems. Since insects accumulate heavy metals predominantly in the gut, it is not surprising that several enzymes in the midgut of L. dispar larvae, including antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT), have been marked as promising biomarkers of Cd presence. Namely, Cd indirectly induces oxidative stress in the cell. However, long-term exposure of the population to pollution often results in increased tolerance and changed sensitivity of biomarkers.
OBJECTIVES:
We aimed to determine specific enzyme activities and isoform patterns of SOD and CAT in the midgut of Lymantria dispar larvae after chronic treatment with Cd. To assess these parameters as biomarkers of Cd exposure, we compared the responses of two populations with different histories of an exposure to pollution.
METHOD / DESIGN:
Egg masses of L. dispar were collected from two localities in Serbia - the uncontaminated forest in Kosmaj Mountain, which is a protected natural resource, and a polluted site near the busy Ibar highway. Larvae were fed wheat germ diet containing 0, 50 or 100 μg Cd/g dry food starting from hatching until they were killed on the 3rd day of the 4th instar. Specific activities of SOD and CAT in the midgut homogenates were determined by spectrophotometric assays. Enzyme isoforms were separated by native polyacrylamide gel electrophoresis. Statistical analyses were performed in GraphPad Prism 7 (GraphPad Software, Inc., USA), where enzyme activities were analyzed by one-way ANOVA followed by Tukey’s post-hoc test. The level of statistical significance was p<0.05.
RESULTS:
Specific activity of SOD was higher in control larvae from the polluted locality compared to the control group from the uncontaminated forest. An exposure to both Cd concentrations decreased SOD activity in larvae from the polluted site. Three SOD isoforms were detected in control groups from both populations. While isoform SOD-2 was absent in the population from Kosmaj after the treatment with higher Cd concentration, both SOD-2 and SOD-3 disappeared in all Cd-treated larvae from the site near the highway. In the population from the unpolluted locality specific activity of CAT was reduced at 100 μg Cd/g dry food, whereas in another population a decrease in enzyme activity was noticed at both Cd concentrations. The same pattern of Cd influence was observed for CAT isoform activity. Only one CAT isoform was present in both control and experimental larvae from both populations.
CONCLUSIONS:
Higher SOD activity in control larvae originating from the site near the highway compared to those from the uncontaminated forest probably indicated the presence of traffic-related pollution that caused oxidative stress. However, neither SOD nor CAT showed activation in response to Cd treatment. A decrease in SOD and CAT activity in both Cd-treated groups in the population from the polluted site could have been a result of the trade-off in favour of the other more efficient defense mechanism(s). Such trade-off might have led to the diminished expression of isoforms SOD-2 and SOD-3. Thus, a decrease in SOD and CAT activities after Cd exposure could be seen as an adaptive strategy of L. dispar population living in chronically polluted habitat. These parameters, with SOD isoform pattern, could be used as biomarkers of Cd exposure in contaminated environments.
PB  - Novi Sad: Faculty of Sciences, University of Novi Sad
C3  - Book of abstracts: The International Bioscience Conference and the 8th International PSU - UNS Bioscience Conference IBSC 2021; 2021 Nov 25-26; Novi Sad, Serbia
T1  - Effects of chronic dietary cadmium on midgut superoxide dismutase (SOD) and catalase (CAT) in larvae from two Lymantria dispar populations
SP  - 77
EP  - 78
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_4912
ER  - 

@conference{
editor = "Mimica-Dukić, Neda, Pajević, Slobodanka, Mandić, Anamarija",
author = "Matić, Dragana and Vlahović, Milena and Ilijin, Larisa and Grčić, Anja and Filipović, Aleksandra and Mrdaković, Marija and Perić Mataruga, Vesna",
year = "2021",
abstract = "INTRODUCTION:
Cadmium (Cd) levels in the environment have increased during decades of intensive industrial development and urbanization. Lymantria dispar has proved to be a suitable organism indicator to monitor Cd pollution in the forest ecosystems. Since insects accumulate heavy metals predominantly in the gut, it is not surprising that several enzymes in the midgut of L. dispar larvae, including antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT), have been marked as promising biomarkers of Cd presence. Namely, Cd indirectly induces oxidative stress in the cell. However, long-term exposure of the population to pollution often results in increased tolerance and changed sensitivity of biomarkers.
OBJECTIVES:
We aimed to determine specific enzyme activities and isoform patterns of SOD and CAT in the midgut of Lymantria dispar larvae after chronic treatment with Cd. To assess these parameters as biomarkers of Cd exposure, we compared the responses of two populations with different histories of an exposure to pollution.
METHOD / DESIGN:
Egg masses of L. dispar were collected from two localities in Serbia - the uncontaminated forest in Kosmaj Mountain, which is a protected natural resource, and a polluted site near the busy Ibar highway. Larvae were fed wheat germ diet containing 0, 50 or 100 μg Cd/g dry food starting from hatching until they were killed on the 3rd day of the 4th instar. Specific activities of SOD and CAT in the midgut homogenates were determined by spectrophotometric assays. Enzyme isoforms were separated by native polyacrylamide gel electrophoresis. Statistical analyses were performed in GraphPad Prism 7 (GraphPad Software, Inc., USA), where enzyme activities were analyzed by one-way ANOVA followed by Tukey’s post-hoc test. The level of statistical significance was p<0.05.
RESULTS:
Specific activity of SOD was higher in control larvae from the polluted locality compared to the control group from the uncontaminated forest. An exposure to both Cd concentrations decreased SOD activity in larvae from the polluted site. Three SOD isoforms were detected in control groups from both populations. While isoform SOD-2 was absent in the population from Kosmaj after the treatment with higher Cd concentration, both SOD-2 and SOD-3 disappeared in all Cd-treated larvae from the site near the highway. In the population from the unpolluted locality specific activity of CAT was reduced at 100 μg Cd/g dry food, whereas in another population a decrease in enzyme activity was noticed at both Cd concentrations. The same pattern of Cd influence was observed for CAT isoform activity. Only one CAT isoform was present in both control and experimental larvae from both populations.
CONCLUSIONS:
Higher SOD activity in control larvae originating from the site near the highway compared to those from the uncontaminated forest probably indicated the presence of traffic-related pollution that caused oxidative stress. However, neither SOD nor CAT showed activation in response to Cd treatment. A decrease in SOD and CAT activity in both Cd-treated groups in the population from the polluted site could have been a result of the trade-off in favour of the other more efficient defense mechanism(s). Such trade-off might have led to the diminished expression of isoforms SOD-2 and SOD-3. Thus, a decrease in SOD and CAT activities after Cd exposure could be seen as an adaptive strategy of L. dispar population living in chronically polluted habitat. These parameters, with SOD isoform pattern, could be used as biomarkers of Cd exposure in contaminated environments.",
publisher = "Novi Sad: Faculty of Sciences, University of Novi Sad",
journal = "Book of abstracts: The International Bioscience Conference and the 8th International PSU - UNS Bioscience Conference IBSC 2021; 2021 Nov 25-26; Novi Sad, Serbia",
title = "Effects of chronic dietary cadmium on midgut superoxide dismutase (SOD) and catalase (CAT) in larvae from two Lymantria dispar populations",
pages = "77-78",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_4912"
}

Mimica-Dukić, N., Pajević, S., Mandić, A., Matić, D., Vlahović, M., Ilijin, L., Grčić, A., Filipović, A., Mrdaković, M.,& Perić Mataruga, V.. (2021). Effects of chronic dietary cadmium on midgut superoxide dismutase (SOD) and catalase (CAT) in larvae from two Lymantria dispar populations. in Book of abstracts: The International Bioscience Conference and the 8th International PSU - UNS Bioscience Conference IBSC 2021; 2021 Nov 25-26; Novi Sad, Serbia
Novi Sad: Faculty of Sciences, University of Novi Sad., 77-78.
https://hdl.handle.net/21.15107/rcub_ibiss_4912

Mimica-Dukić N, Pajević S, Mandić A, Matić D, Vlahović M, Ilijin L, Grčić A, Filipović A, Mrdaković M, Perić Mataruga V. Effects of chronic dietary cadmium on midgut superoxide dismutase (SOD) and catalase (CAT) in larvae from two Lymantria dispar populations. in Book of abstracts: The International Bioscience Conference and the 8th International PSU - UNS Bioscience Conference IBSC 2021; 2021 Nov 25-26; Novi Sad, Serbia. 2021;:77-78.
https://hdl.handle.net/21.15107/rcub_ibiss_4912 .

Mimica-Dukić, Neda, Pajević, Slobodanka, Mandić, Anamarija, Matić, Dragana, Vlahović, Milena, Ilijin, Larisa, Grčić, Anja, Filipović, Aleksandra, Mrdaković, Marija, Perić Mataruga, Vesna, "Effects of chronic dietary cadmium on midgut superoxide dismutase (SOD) and catalase (CAT) in larvae from two Lymantria dispar populations" in Book of abstracts: The International Bioscience Conference and the 8th International PSU - UNS Bioscience Conference IBSC 2021; 2021 Nov 25-26; Novi Sad, Serbia (2021):77-78,
https://hdl.handle.net/21.15107/rcub_ibiss_4912 .