Janja Horvatić

Changes in N and P limitation induced by water level fluctuations in Nature Park Kopački Rit (Croatia): nutrient enrichment bioassay

The importance of nutrients as limiting factors might vary in environments with different limnological characteristics. In this article we evaluate the effect of frequency and duration of flooding on nutrient limitation in a riparian floodplain. Variations in N and P limitations were studied in the period of low (2003) and high (2004) water level in two different floodplain habitat types in the Nature Park Kopački Rit (Croatia), a floodplain area of the Danube River. In 2003 and 2004, the limnological characteristics of floodplain lake (Lake Sakadaš) and the channel (Stara Drava) differed due to their hydrological regimes. Potential for nutrient limitation was determined by DIN:TP and TN:TP ratios, while the actual nutrient limitation was assessed by nutrient enrichment bioassay. A change from non-limited to N-limited conditions in the channel, and consistent actual N limitation in the lake was determined by the nutrient enrichment experiment. Of the two ratios, DIN:TP matched better with the bioassay data. Phosphorus limitation was only occasionally evident. Changes in trophic conditions from hypereutrophy to eutrophy (between low and high water levels) reflected the importance of the hydrologic regime as a factor which can modify the trophic state of Lake Sakadaš. Compared with 2003, the increase of total nitrogen concentrations in 2004 calls attention to the importance of nitrogen inputs from the Danube to the system with excessive phosphorus concentrations.

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.

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 Buško 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 Buško 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.

Spatial Distribution of Nutrient Limitation in the Danube River Floodplain in Relation to Hydrological Connectivity

In this paper, we describe the influence of flooding on nutrient conditions and phytoplankton Chl-a concentrations in a hydrologically dynamic Danube floodplain in North–Eastern Croatia. Based on hydrological typology, the connectivity gradient extends from the main river stream (Eupotamal) to the backwater channels and side-arms (Parapotamal) to the edge of the floodplain (Paleopotamal). A spatial gradient of phytoplankton Chl-a occurred during low water levels, while uniformity of Chl-a across wetland habitats was observed during high water levels. Nutrient enrichment assays indicated that inorganic nutrients in Eupotamal waters (N/P ratio 16–22) could support a higher growth rate of the assay alga Chlorella kessleri, while potential N limitation was found in Parapotamal and Paleopotamal waters (both with N/P ratios < 14). Spatial expansion of N limitation toward the main river was observed as inundation continued. Lateral flood pulse connectivity along with the duration of inundation period affects the input and the retention of nutrients, especially of nitrogen. Understanding of the relationship between spatial (location of the floodplain waterbodies regarding the river) and temporal (hydrological connectivity between the river and its floodplain) dimensions contribute to the importance of healthy floodplains for the maintenance of water quality and environmental conditions in the Danube River.

Short term exposure of Lemna minor and Lemna gibba to mercury, cadmium and chromium

The effects of mercury (Hg), cadmium (Cd) and chromium (Cr) in concentrations ranging from 0.02 to 20 mg L−1 applied for 24 h were assessed in Lemna minor and Lemna gibba by measuring changes in protein concentration, ascorbic acid, phenolics, malondialdehyde (MDA), hydrogen peroxide (H2O2), the activity of guaiacol peroxidase (G-POX) and catalase (CAT). Ascorbic acid, phenolics, catalase and guaiacol peroxidase played a key role in the antioxidative response of L. gibba. Inadequate activity of antioxidant enzymes in the L. minor resulted in MDA and H2O2 accumulation. In both used species, Hg treatment decreased protein content and increased CAT and G-POX activity, but decreased MDA and H2O2 levels. Cadmium and chromium had opposite impacts on two used Lemna species on almost all observed parameters. Enhanced antioxidative responses of L. gibba to lower concentrations of Hg, Cd and Cr indicated greater abiotic stress tolerance than L. minor.

Distribution and morphological variations of invasive macrophytes Elodea nuttallii (Planch.) H. St. John and Elodea canadensis Michx in Croatia

Abstract The invasive species Elodea nuttallii was recorded for the first time in the Croatian flora in 2006, in the drainage channels of Kopački rit (Baranja). After its establishment, E. nuttallii begins to spread to the eastern and northern part of the drainage channel network from 2006-2009. High water levels are responsible for the linear spreading direction of E. nuttallii, E. nuttallii and E. canadensis show a wide range of morphological variation, which is characteristic of successful invaders. To show morphological variations of two Elodea species, the most important characters indicated in the literature were measured on 24 fresh collected samples from the seven sites in Croatia. In spite of some overlap in leaf length and width between the two Elodea species, the differences of all morphological traits except internode length are statistically significant. In E. nuttallii leaf width, length and internode length show a higher morphological variability as a result of the higher adaptive strategy to environmental parameters. The most reliable morphologi-cal characters distinguishing E. nuttallii and E. canadensis are leaf width 0.5 mm below the tip and the angle at the apex. E. nuttallii can be expected to spread to other areas of Croatia.

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.

Trophic State and Water Quality in the Danube Floodplain Lake (Kopački Rit Nature Park, Croatia) in Relation to Hydrological Connectivity

The investigated floodplain lake is located in the marginal part of the Kopacki Rit Nature Park, Croatia, a floodplain area of the Danube. In this chapter, we have assessed the influence of hydrological connectivity with the main channel on the trophic state and water quality in the Lake Sakadas. The evaluation of trophic state combines a number of physical, chemical and biological parameters with the objective to select the relevant indicators for the floodplain lakes. The following environmental parameters have been considered: water temperature, nutrients, dissolved oxygen, water transparency, chlorophyll-a concentration, phytoplankton primary productivity, benthic and periphytic communities and laboratory experiments (nutrient enrichment bioassay). An integrative approach, which includes in situ research and laboratory experiments, indicated strong correlations of abiotic and biotic components with a hydrological regime. Also, during the biological monitoring, occurrence and spread of invasive species was noted as an indication of ecosystem disturbance (hydrological and/or anthropogenic). Therefore, our results contribute to the perception of importance of the hydrological connectivity (by inundation) on the eutrophication process in the investigated floodplain lake.