Physiological Response of Nutrient-Stressed Lemna gibba to Pulse Colloidal Silver Treatment.

Wastewater is a source of many environmental pollutants and potentially high concentrations of essential plant nutrients. Site-specific nutrient levels may influence the response of exposed plants to a chemical stressor. In the present study, we focused on the responses of model aquatic macrophyte swollen duckweed (Lemna gibba L.) to a short pulse exposure and a commercially available colloidal silver product as a potential environmental chemical stressor, combined with two levels of total nitrogen and phosphorus nutrition. Treatment with the commercially available colloidal silver product caused oxidative stress in L. gibba plants under both high and low nutrient levels. Plants grown and treated under high nutrient levels showed lower levels of lipid peroxidation and hydrogen peroxide accumulation, as well as higher levels of photosynthetic pigment content in comparison to treated plants under low nutrient levels. Higher free radical scavenging activity for plants treated with silver in combination with high nutrient levels resulted in better overall protection from silver-induced oxidative stress. The results showed that external nutrient levels significantly affected the L. gibba plant's response to the colloidal silver presence in the environment and that nutrient levels should be considered in the assessment of potential environmental impact for contaminants.

Growth inhibition and recovery patterns of common duckweed Lemna minor L. after repeated exposure to isoproturon.

Aquatic non-targeted organisms are more likely to be exposed to herbicides in multiple pulse events then long continuous exposure. The potential of an organism to recover between exposures has an important role in the overall effects of the toxicant. Common duckweeds show high potential for recovery after a single exposure to isoproturon. To evaluate the growth patterns and recovery potential between multiple exposures, L. minor plants were exposed to isoproturon in three repetitive 7-day treatment cycles in three time-variable exposure scenarios with equivalent time-weighted average concentrations. The growth was significantly inhibited during each exposure phase with significant cumulative effects in every subsequent treatment cycle resulting in a cumulative decrease in biomass production. However, inhibitory effects were reversible upon transferring plants to a herbicide-free nutrient solution. These results indicate that L. minor plants have a high recovery potential even after multiple exposures to isoproturon. Observed cumulative decrease in biomass production, as well as the potential for fast and efficient recovery from repeated herbicide exposure, might affect the competitiveness of L. minor in surface water communities. The observations made during each exposure period, recovery patterns, and the resulting cumulative effects over time may contribute to further development, calibration and validation of mechanistic toxicokinetic/toxicodynamic models for simulating the effects of pesticides on aquatic plants populations in the laboratory and environmental conditions.

Phytotoxicity assessment of isoproturon on growth and physiology of non-targeted aquatic plant Lemna minor L. - A comparison of continuous and pulsed exposure with equivalent time-averaged concentrations.

Phenylurea herbicides are often present in the aquatic ecosystems and may be accumulated by the non-targeted organisms and impose a negative effect on the organism and the community. This study aims to investigate and compare the effects of two different isoproturon (IPU) pulse exposure scenarios on the non-targeted aquatic plant Lemna minor with effects observed in the standard test with continuous exposure. The obtained results showed that continuous IPU treatment causes significant reduction of photosynthetic pigment concentration and proteins as well as inhibition of L. minor growth. The activities of CAT, G-POX, and APX were significantly induced to diminish the accumulation of ROS under IPU treatment, but the induction of antioxidant enzymes was not sufficient to protect the plants from herbicide-induced oxidative stress. The growth of L. minor under pulse exposure to IPU recovers fast, but pulse treatment results in significant physiological changes in treated plants. The accumulation of H2O2 and lipid peroxidation products, alongside the reduced concentration of proteins and photosynthetic pigments in pulse treatment after a recovery period, indicates that IPU causes prolonged oxidative stress in L. minor plants. The recovery potential of L. minor plants after treatment with herbicides may have an important role in maintaining the population of essential primary producers in aquatic ecosystems, but IPU-induced physiological changes could potentially have a significant role in modulating the response of the plants to the next exposure event.

Physiological and biochemical effect of silver on the aquatic plant Lemna gibba L.: Evaluation of commercially available product containing colloidal silver.

This paper aims to evaluate the effects of a product containing colloidal silver in the aquatic environment, using duckweed Lemna gibba as a model plant. Therefore, growth parameters, photosynthetic pigments content and protein content as physiological indices were evaluated. Changes in the content of non-enzymatic antioxidants and activity of several antioxidant enzymes, alongside with the accumulation of hydrogen peroxide and lipid peroxidation end-products were assessed to explore the potential of colloidal silver to induce oxidative stress. The commercially available colloidal silver product contained a primary soluble form of silver. The treatment with colloidal silver resulted in significant physiological and biochemical changes in L. gibba plants and a consequent reduction of growth. Accumulation of silver caused altered nutrient balance in the plants as well as a significant decrease in photosynthetic pigments content and protein concentration. The antioxidative response of L. gibba plants to treatment with colloidal silver was inadequate to protect the plants from oxidative stress caused by metal accumulation. Silver caused concentration-dependent and time-dependent hydrogen peroxide accumulation as well as the elevation of lipid peroxidation levels in L. gibba plants. The use of commercially available products containing colloidal silver, and consequent accumulation of silver, both ionic and nanoparticle form in the environment, represents a potential source of toxicity to primary producers in the aquatic ecosystem.

S-metolachlor promotes oxidative stress in green microalga Parachlorella kessleri - A potential environmental and health risk for higher organisms.

The estimation of the toxic influences of herbicide products on non-target aquatic organisms is essential for evaluation of environmental contamination. We assessed the effects of the herbicide S-metolachlor (S-MET) on unicellular green microalga Parachlorella kessleri during 4-72 in vitro exposure to concentrations in the range 2-200µg/L. The results have shown that S-MET had a significant effect on algae, even in doses 10 and 20 times lower than the EC50 values obtained for P. kessleri (EC50-72h=1090µg/L). It generates reactive oxygen species in algae, decreases their growth and photosynthetic pigment concentration, changes their ultrastructure and alters the cellular antioxidant defence capacities. The levels of protein adducts with the reactive aldehyde 4-hydroxy-2-nonenal (HNE), the end-product of lipid peroxidation, were significantly elevated in S-MET treated cells revealing the insufficient effectiveness of P. kessleri antioxidant mechanisms and persistent lipid peroxidation. Since algae are fundamental aquatic food component, the damaged algal cells, still capable of dividing while having persistently increased content of HNE upon S-MET contamination could represent an important environmental toxic factor that might further affect higher organisms in the food chain.

Distribution and age-related bioaccumulation of lead (Pb), mercury (Hg), cadmium (Cd), and arsenic (As) in tissues of common carp (Cyprinus carpio) and European catfish (Sylurus glanis) from the Busko Blato reservoir (Bosnia and Herzegovina).

The purpose of this study was to quantify the bioaccumulation of Pb, Hg, Cd, and As in tissues of carp (Cyprinus carpio) and catfish (Silurus glanis) from Busko Blato in Bosnia and Herzegovina. Arsenic concentrations were below the Maximal Admissible Concentration (MAC) for Croatia and other countries. Mercury concentrations were below 1 mg kg(-1), but in most muscle samples of both species and all catfish liver samples, the values were higher than 0.5 mg kg(-1) (higher than the MAC for many countries including Croatia). Lead concentrations were higher than 1 mg kg(-1) (the MAC for Croatia) in most muscle samples; all kidney and most catfish liver samples also exceeded 1 mg kg(-1). Cadmium concentrations in all tissues, other than the gonads, were higher than 0.1 mg kg(-1) (MAC for Croatia), with the highest concentrations found in the kidneys. The only gender difference was found in carp, where a 68.4% higher concentration of As was found in the fry compared to the milt (P<0.05). Concentrations of all of the elements were higher in catfish compared to carp for most tissues. Significant correlations were found between all of the elements in the muscles and the liver of carp. In catfish, the muscles were the only tissue in which multiple correlations were found. Linear positive correlations with age and body mass were demonstrated for the concentrations of all heavy metals for all tissues except the gonads in both fish species. We concluded that significant heavy metal accumulation in carp and a catfish tissues correlates with age and body mass; bioaccumulation is species- and tissue-specific and is different for each element.

Endogenous 4-hydroxy-2-nonenal in microalga Chlorella kessleri acts as a bioactive indicator of pollution with common herbicides and growth regulating factor of hormesis.

Oxidative stress, i.e. excessive production of reactive oxygen species (ROS), leads to lipid peroxidation and to formation of reactive aldehydes (e.g. 4-hydroxy-2-nonenal; HNE), which act as second messengers of free radicals. It was previously shown that herbicides can induce ROS production in algal cells. In the current paper, the unicellular green microalga Chlorella kessleri was used to study the effect of two herbicides (S-metolachlor and terbuthylazine) and hydrogen peroxide (H(2)O(2)) on oxidative stress induction, HNE formation, chlorophyll content and the cell growth. Production of HNE was detected in this study for the first time in the cells of unicellular green algae using the antibody specific for the HNE-histidine adducts revealing the HNE-histidine adducts even in untreated, control C. kessleri. Exposure of algal cells to herbicides and H(2)O(2) increased the ROS production, modifying production of HNE. Namely, 4h upon treatment the levels of HNE-histidine conjugates were below controls. However, their amount increased afterwards. The increase of HNE levels in algae was followed by their increased growth rate, as was previously described for human carcinoma cells. Hence, changes in the cellular HNE content upon herbicide treatment inducing lipid oxidative stress and alterations in cellular growth rate of C. kessleri resemble adaptation of malignant cells to the HNE treatment. Therefore, as an addition to the standard toxicity tests, the evaluation of HNE-protein adducts in C. kessleri might indicate environmental pollution with lipid peroxidation-inducing herbicides. Finally, C. kessleri might be a convenient experimental model to further study cellular hormetic adaptation to oxidative stress-derived aldehydes.

Heavy metal distribution in tissues of six fish species included in human diet, inhabiting freshwaters of the Nature Park "Hutovo Blato" (Bosnia and Herzegovina).

The aim of the study was to quantify heavy metal (mercury, lead, cadmium, and arsenic) concentration in tissues (muscles, liver, kidney, gills, and gonads) of six fish species (carp: Cyprinus carpio, tench: Tinca tinca, pumpkinseed: Lepomis gibosus, prussian carp: Carassius auratus gibelio, hasselquist: Salmo dentex, eel: Anguilla anguilla) from the freshwaters of the Nature Park Hutovo Blato, Bosnia and Herzegovina, and determine whether they are potentially harmful for human health if included in the diet. Fish were angled from the Svitava Lake in the second part of August of the year 2003, and fish tissues were stored at -18 degrees C until analysis. Heavy metal concentration was determined by atomic absorption spectrophotometry in the Veterinary Institute Brno, Czech Republic, and expressed as mg.kg(-1) of wet tissue. Concentration of mercury, lead, and arsenic in most tissues of all analyzed fish types is lower than the maximal allowed concentration (MAC) in most countries. Cadmium concentration is also low in muscles and gonads, but kidney, liver, and gill concentrations exceed MAC value in most countries. Hasselquist, an endemic type for that region, differs from other fish types in the fact that it has very low cadmium concentration in liver and kidney, but the highest concentration of arsenic in most tissues, especially muscles. In muscles and gonads of all fish types analyzed, Pb is present in higher concentration than Cd, whereas in liver, gills, and particularly kidney, the situation is opposite, suggesting diverse metabolic pathways and unequal bioaccumulation of these two metals in different fish tissues. Although the region of the Nature Park Hutovo Blato in Bosnia and Herzegovina is not an agricultural territory, the intensive agricultural activities in the neighboring regions already result in high cadmium concentration in inner organs of fish species analyzed. Therefore, fish types in the freshwaters of the Park may be included in the human diet, but without inner organs and gills (or the whole head).

Heavy metal concentration in fish tissues inhabiting waters of "Busko Blato" reservoir (Bosnia and Herzegovina).

Heavy metals concentration (mercury, lead, cadmium, arsenic, copper, zinc and chromium) in tissues (muscles, liver, kidney and gonads) of Dalmatian barbelgudgeon, the nase, the souffie and brown trout, inhabiting waters of Busko Blato reservoir in Bosnia and Herzegovina, has been determined by atomic absorption spectrophotometry. The meat of the tested fish sorts does not contain elevated concentration of most analyzed heavy metals with exception of lead (higher than MAC in Italy, Germany and Denmark) and mercury (in muscles of brown trout higher than MAC in most countries). The lowest level of all heavy metals is always detected in gonads, with higher values in fry compared to milt for copper, zinc, chromium and arsenic. The highest copper concentration is observed in the liver from the souffie which is suggested as a suitable biomonitor for copper intoxication. In muscles of all fish sorts, lead was always present in much higher concentration than cadmium, while in kidneys of most fish sorts, lead and cadmium concentrations were similar. We showed that bioaccumulation of some heavy metals in the fish sorts analyzed is tissue and sex dependent. Also, we concluded that the small water exchange in reversible shallow reservoir does not induce elevated concentration of heavy metals in fish tissues inhabiting Busko Blato.