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The effects of different concentrations of benzo(a)pyrene (BaP) (2, 10, 20, 100, 200, and 2000 ng/g dry food weight) on the activity of acetylcholinesterase (AChE) and concentration of heat-shock protein 70 (hsp70) in fifth instar Lymantria dispar brain tissue were investigated. Inhibition of AChE is a general effect biomarker at the individual level. We observed the most efficient inhibition of this enzyme at medium and high BaP concentrations (20, 100, 200, and 2000 ng/g dry food weight). Western blot analysis revealed the presence of two hsp 70 isoforms, with changed amounts depending on the BaP concentration. A fluctuating response in hsp70 concentration to ingested BaP was observed: with decreased levels in the groups fed 2 and 100 ng BaP/g dry food weight, but increased concentrations in the groups given 10 and 2000 ng/g dry food weight. Even though we detected changes in AChE activity and hsp70 concentration in L. dispar brain tissue upon exposure to different BaP concentrations, we cannot characterize them as sensitive and reliable biomarkers for this xenobiotic in L. dispar caterpillars.

This study aimed to examine the effects of ubiquitous polycyclic aromatic hydrocarbon fluoranthene, supplemented to an artificial diet, on the fitness-related traits and activity of midgut antioxidative enzymes-superoxide dismutase (SOD) and catalase (CAT), and expression of their isoforms in the fifth-instar gypsy moth Lymantria dispar L. Prolonged duration of development and reduced weight and relative growth rate were recorded in larvae reared on the diets supplemented with different concentrations of fluoranthene. SOD and CAT activities were significantly higher in the midguts of fluoranthene-treated larvae, compared to that of the control group. Different expression patterns were detected for SOD as well as for CAT isoforms, depending on the supplemented concentration of fluoranthene. Obtained results suggest that the activity of these enzymes in gypsy moth larvae may be used as biomarkers for assessing pollution, even at low concentrations of the pollutant.

Increased presence of benzo[a]pyrene in the environment underlines the need for development of sensitive biomarkers for monitoring. Antioxidative enzymes could be used as early-warning signals because of their sensitivity and applicability. The activity of 2 antioxidative enzymes, superoxide dismutase (SOD) and catalase (CAT), were measured in midgut tissues of fifth instar Lymantria dispar larvae exposed to different concentrations of benzo[a]pyrene: 2 ng, 10 ng, 20 ng, 100 ng, 200 ng, and 2000 ng benzo[a]pyrene/g dry food weight. Larval development, larval mass, and relative growth rate were also monitored. The authors detected prolonged larval development, as well as reduced larval mass and relative growth rate in larvae exposed to all benzo[a]pyrene concentrations. The L. dispar midgut SOD activity was significantly increased, and 2 SOD isoforms were detected on native polyacrylamide gel electrophoresis in larvae fed on artificial diet supplemented with benzo[a]pyrene. In contrast, the control group had only 1 isoform. Catalase activity was significantly increased in all benzo[a]pyrene-treated larvae. Native gel electrophoresis showed that a switch in active CAT isoforms occurred after benzo[a]pyrene treatment. Thus, SOD and CAT in polyphagous herbivorous L. dispar larvae are very sensitive to low concentrations of benzo[a]pyrene. Therefore, they could be used as biomarkers for exposure and effects of this toxic polycyclic aromatic hydrocarbon.

α-Amylase production by solid-state fermentation of different Bacillus sp. was studied previously on different fermentation media. However, no study has been reported on the influence of selected media on expression of desired amylase isoforms such as raw-starch-digesting amylase (RSDA). In this paper, the influence of different inexpensive and available agro-resources as solid media (corn, wheat and triticale) on α-amylase isoform induction from three wild-type Bacillus sp., selected among one hundred strains tested, namely 9B, 12B and 24A was investigated. For all three strains, tested amylases were detected in the multiple forms; however, number and intensity of each form differed depending on the solid media used for growth. To determine which isoform from Bacillus sp. 12B was RSDA, the suspected isoform was purified. The optimum pH for the purified α-amylase isoform was 6.0-8.0, while the optimum temperature was 60-90 °C. Isoform was considerably thermostable and Ca(2+)-independent, and actually the only α-amylase active towards raw starch. Purification and characterization of RSDA showed that not all of the solid media tested induced RSDA. From an economic point of view, it might be significant to obtain pure isoenzyme for potential use in the raw-starch hydrolysis, since it was 5 times more efficient in raw corn starch hydrolysis than the crude amylase preparation.

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The release of neurosecretory material from A2 neurosecretory neurons (NSNs) was stimulated in Lymantria dispar fourth instar caterpillars exposed to a temperature of 35 °C for 1, 12, and 24 h, as well as those allowed recover after exposure (12 h at 35 °C, then 12 h at 23 °C). The levels of 2 protein forms with the same molecular mass as bombyxin (3-4 and 4-5 kDa) increased with prolonged exposure to 35 °C. The second band was present only in the groups exposed to this stressor. There was intensified synthetic activity and a low level of secretion in L2' NSNs after exposure to 35 °C. We previously found these NSNs to be immunopositive for prothoracicotropic neurohormone. After this stress, densitometric analysis revealed a decreased amount of the 11-12 kDa isoform (present in the control group). The new isoform (13-15 kDa), expressed after exposure of the insects to a high temperature, increased in amount with prolonged exposure and after recovery at 23 °C. Short-term exposure of caterpillars to high temperatures (35 °C) is a stressor and activates carbohydrate metabolism, while PTTH immunopositive NSNs are secretory-inactive during acute thermal stress regimes.