Management of inorganic elements by overwintering physiology of cold hardy larvae of European corn borer (Ostrinia nubilalis, Hbn.).

The European corn borer (Ostrinia nubilalis, Hbn.), enters diapause, a strategy characterized by arrest of development and reproduction, reduction of metabolic rate and the emergence of increased resistance to challenging seasonal conditions as low sub-zero winter temperatures. The aim of this study was to investigate the potential role of inorganic elements in the ecophysiology of O. nubilalis, analysing their content in the whole body, hemolymph and fat body, both metabolically active, non-diapausing and overwintering diapausing larvae by ICP-OES spectrometer following the US EPA method 200.7:2001. O nubilalis as many phytophagous lepidopteran species maintain a very low extracellular sodium concentration and has potassium as dominant cation in hemolymph of their larvae. Changes in hemolymph and the whole body sodium content occur already at the onset of diapause (when the mean environmental temperatures are still high above 0 ºC) and remain stable during the time course of diapause when larvae of this species cope with sub-zero temperatures, it seems that sodium content regulation is rather a part of diapausing program than the direct effect of exposure to low temperatures. Compared to non-diapausing O. nubilalis larvae, potassium levels are much higher in the whole body and fat body of diapausing larvae and substantially increase approaching the end of diapause. The concentration of Ca, Mg, P and S differed in the whole body, hemolymph and fat body between non-diapausing and diapausing larvae without a unique trend during diapause, except an increase in their contents at the end of diapause.

Identification of a metallothionein gene and the role of biological thiols in stress induced by short-term Cd exposure in Ostrinia nubilalis.

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.

Phytoremediation potential of the naturally occurring wetland species in protected Long Beach in Ulcinj, Montenegro.

Long Beach, situated in southern Montenegro, is subject to considerable biogenic and abiogenic influences. Thus, analyzing total heavy metal content in soil and plants in this region is, while challenging, highly important in order to assess the level for determining the soil degradation level and the phytoremediation potential of naturally growing salt marsh species. This area together with a Bojana river and backshore forms a real vegetation mosaic where habitats of various types coexist. Therefore, it represents good model system. In the present study, the levels of As, Al, B, Cd, Co, Cr, Cu, Hg, Fe, Mn, Mo, Ni, Pb and Zn in coastal soils as well as in eight salt marsh plants: Bolboschoenus maritimus, Juncus acutus, Juncus anceps, Juncus articulatus, Juncus gerardii, Juncus maritimus, Scirpus holoschoenus and Schoenus nigricans, were investigated in order to identify the plant species that can be used for the remediation of polluted sites, especially those located along the coastline. The obtained results show that species J. gerardii, J. articulatus and B. maritimus can be clearly separated from J. acutus, J. anceps, J. maritimus, S. holoschoenus and Sh. nigricans based on the degree of heavy metal accumulation in various organs. Moreover, analyses revealed that the bioaccumulation factor of underground organs is significantly higher relative to that of the aboveground parts for almost all investigated metals and species. The bioaccumulation factor had the highest value in the underground organs of J. gerardii and B. maritimus, where a value of 3.37 was measured for B and 2.54 for Hg, respectively. Hence, as all investigated species are "underground accumulators" for most of the analyzed metals, they could be useful for phytostabilization and phytoremediation of B and Hg in particular. Moreover, each plant species can be used in the phytoremediation process targeting specific heavy metals.

The effect of long term exposure to cadmium on Ostrinia nubilalis growth, development, survival rate and oxidative status.

In this study the effect of long term exposure to cadmium (Cd) on Ostrinia nubilalis larval growth, development, survival rate and oxidative status was analyzed. Newly hatched first instar - L1 larvae were reared on a Cd contaminated diet until the larvae reached the final, fifth instar - L5 or developed into pupae. In total, six experimental groups, five treatments (concentrations of Cd in fresh diet: Cd I: 0.73, Cd II: 3.70, Cd III: 6.85, Cd IV: 41.71 and Cd V: 77.53 mg kg-1) and a control group (C) were set up. The results of the experiment showed that exposure to higher concentrations of Cd (41.71 and 77.53 mg kg-1) had a significant influence on development and redox status of O. nubilalis larvae: (1) the development rate was strongly reduced resulting in a prolonged pupation time; (2) the survival rate of larvae was prominently lower; (3) bioaccumulation factor (measured in pupae) was reduced which indicated that larvae could accumulate Cd to a certain level; (4) the level of the lipid peroxidation was significantly higher, which points to oxidative damage; (5) the expression of Mtn was significantly up-regulated while Cat and GPx genes down-regulated. In conclusion, long term exposure to dietary Cd in a concentration of 41.7 mg kg-1 and higher, induced oxidative stress and slowed down growth and development of O. nubilalis larvae.

Mercury content in agricultural soils (Vojvodina Province, Serbia).

The Vojvodina Province in northern Serbia is well known for its intensive field crops production. Over 90 % of total arable land, which represents more than 1500.000 ha, is used for field or vegetable crop production. A grid superimposed on Vojvodina land by means of a GIS tool (GIS ArcView 10) has divided land into 4 × 4 km units, each representing an area of 1600 ha. Total number of 1370 bulked soil samples were taken (0-30 cm depth) from agricultural land and analysed for total mercury content THg. The samples were analysed using Direct Mercury Analyzer DMA 80 Milestone. Quality control was carried out with IRMM BCR reference materials 143R. The aim of this study was to determine the total content of Hg in agricultural soils and its spatial distributions in different parts of Vojvodina Province. The obtained results were within interval 0.008-0.974 mg kg-1. The average concentration of Hg was 0.068, with median 0.048 mg kg-1. The correlation was determined between Hg concentration and organic matter content in the soil. Content of Hg coincides with main geomorphological units of Vojvodina Province. Average values of Hg concentrations for soils formatted on different geomorphological units were 0.031 for sandy area with dune fields, 0.048 for alluvial terraces, 0.055 for upper Pleistocene terraces, 0.058 for loess plateaus, 0.083 for mountains and 0.092 mg kg-1 for alluvial plains. Hg spatial distribution confirmed that most of Vojvodina Province area has geochemical origin of Hg. Higher concentration of Hg on alluvial plains indicated that the origin of Hg near rivers could be from anthropogenic source. The main rivers in Vojvodina have been dammed more than a century ago. Thus, higher concentrations of Hg in their alluvial plains out of narrow dammed zone around the rivers must be related to natural and anthropogenic sources located in their huge catchments. Higher content of Hg in mountain region can be explained by high clay content in these soils. Additional hotspots of Hg concentration of top soils are related to geographical locations of major towns. The obtained results also indicated that the measured levels of Hg in the soil are not limiting factors for production of safe food in Vojvodina.

Correlation between radioactivity levels and heavy metal content in the soils of the North Kosovska Mitrovica environment.

This paper reports the results of radioactivity concentrations and heavy metal content in soil samples collected from non-agricultural areas in the municipality and vicinity of North Kosovska Mitrovica, formerly the most important mining area in Europe. The estimated average activity concentrations of (226)Ra, (232)Th, (40)K and (137)Cs are 40.6 ± 19, 48 ± 25.4, 743.2 ± 200.5 and 81 ± 119 Bq kg(-1), respectively. The estimated average absorbed dose rate in the air and the annual effective dose are 78.7 nGy h(-1) and 96.6 µSv, respectively. The radium equivalent activity and external hazard index were also calculated. High contents of Pb, Zn, Cu, Cd, As and Ni were found in the analyzed soil samples, thus indicating pollution of the environment. Most metals have a wide range of values spanning 2 to 3 orders of magnitude, which is particularly evident for Pb and Zn. Correlations between the activity concentrations of the radionuclides, the heavy metal content in soil and the basic soil properties were determined by means of the Pearson linear coefficient. Strong positive correlations between the naturally occurring radionuclides, and also among Pb, Zn, Cu and Cd were found.

Metal contamination of short-term snow cover near urban crossroads: correlation analysis of metal content and fine particles distribution.

Snow samples were collected near crossroads in Novi Sad, Serbia, during December 2009 to assess metal concentrations (Ca, K, Zn, Fe, Cu, Mn, Al, Pb and Na), fine particle distribution and pH value. The filtered samples of melted snow were analysed, with a focus on particles smaller than a few µm. The most common values of the frequency number distribution curve were either in the range of 0.05-0.07 µm or one order of magnitude higher (0.2-0.5 µm). At examined locations metal concentrations varied from 0.0004 mg L(-1) for Pb to 18.9 mg L(-1) for Na. Besides Na, which mostly originated from de-icing salt, Ca is found to be the most abundant element in snow indicating the dominant influence of natural factors on snow chemistry. No significant difference was found in concentration of the elements at locations near crossroads with either low or high traffic volume, except for Na and Zn. To investigate how metals were related a correlation analysis was done for the concentrations of metals and with respect to the particle size distribution parameters and pH values. The major influence on the fine particle volume (mass) was concluded to be due to the elements from anthropogenic sources. This conclusion was based on the significant positive correlation between Fe, Zn and Al and the fine particle volume based distribution parameters.