Soil copper uptake by land snails: A semi-field experiment with juvenile Cantareus aspersus snails.

There is currently limited scientific evidence linking soil copper and land snails, although these invertebrates are important players in terrestrial ecosystems. In the present study, Cantareus aspersus juveniles, were exposed in two successive phases of 30 days each, to soil spiked with increasing concentrations of copper sulfate. Copper concentrated preferentially and in a dose-dependent manner in the hepatopancreas. In the case of specimens previously exposed to Cu-spiked soils, Cu retention kinetics were independent from the effects of a new exposure event. There was no effect on shell growth, but significant mortality was observed at 60 days. The no observed effect concentration and the lowest observed effect concentration for mortality in snails, were ˜ 41 and 54 mg, respectively, per grams dry weight in the hepatopancreas. The results demonstrate, for the first time, that terrestrial gastropods can accumulate soil Cu autonomously from dietary uptake.

Impact of soil cadmium on land snails: a two-stage exposure approach under semi-field conditions using bioaccumulative and conchological end-points of exposure.

Land snails are highly tolerant to cadmium exposure and are able to accumulate soil cadmium independently of food ingestion. However, little information exists on the kinetics of cadmium retention in terrestrial gastropods exposed to an increase in the soil cadmium content, over time. There is also little knowledge about how exposure to cadmium-polluted soils influences shell growth and architecture. In this context, we examined cadmium accumulation in the hepatopancreas and shell of juvenile Cantareus aspersus exposed to elevating high levels of cadmium in soil. Also, the toxicity of cadmium to snails was assessed using a range of conchological endpoints, including shell height, width, volume, allometry and integrity. Test snails, aged three months, were reared under semi-field conditions, fed an uncontaminated diet and exposed first, for a period of 30 days, to a series of soil cadmium concentrations, and then, for a second period of 30 days, to soils with higher cadmium content. Cadmium showed a dose-dependent accumulation in both the hepatopancreas and shell. The kinetics of cadmium retention in the hepatopancreas of snails previously exposed to cadmium-spiked soils was significantly influenced by a new exposure event. The shell was not a relevant bioaccumulator for soil cadmium. Under the present experimental conditions, only high cadmium exposure significantly affected either the shell growth or snail survival. There was no consistent effect on shell allometry, but the shell integrity, especially in rapidly growing parts, appeared to be affected by high cadmium exposure. Our results attest to the value of hepatopancreas for describing cadmium retention in land snails and to the difficulty of using conchological parameters in field surveys for estimating the environmental hazard of soil cadmium.

Soil manganese enrichment from industrial inputs: a gastropod perspective.

Manganese is one of the most abundant metal in natural environments and serves as an essential microelement for all living systems. However, the enrichment of soil with manganese resulting from industrial inputs may threaten terrestrial ecosystems. Several studies have demonstrated harmful effects of manganese exposure by cutaneous contact and/or by soil ingestion to a wide range of soil invertebrates. The link between soil manganese and land snails has never been made although these invertebrates routinely come in contact with the upper soil horizons through cutaneous contact, egg-laying, and feeding activities in soil. Therefore, we have investigated the direct transfer of manganese from soils to snails and assessed its toxicity at background concentrations in the soil. Juvenile Cantareus aspersus snails were caged under semi-field conditions and exposed first, for a period of 30 days, to a series of soil manganese concentrations, and then, for a second period of 30 days, to soils with higher manganese concentrations. Manganese levels were measured in the snail hepatopancreas, foot, and shell. The snail survival and shell growth were used to assess the lethal and sublethal effects of manganese exposure. The transfer of manganese from soil to snails occurred independently of food ingestion, but had no consistent effect on either the snail survival or shell growth. The hepatopancreas was the best biomarker of manganese exposure, whereas the shell did not serve as a long-term sink for this metal. The kinetics of manganese retention in the hepatopancreas of snails previously exposed to manganese-spiked soils was significantly influenced by a new exposure event. The results of this study reveal the importance of land snails for manganese cycling in terrestrial biotopes and suggest that the direct transfer from soils to snails should be considered when precisely assessing the impact of anthropogenic Mn releases on soil ecosystems.

A novel exploratory chemometric approach to environmental monitorring by combining block clustering with Partial Least Square (PLS) analysis.

BACKGROUND: Given the serious threats posed to terrestrial ecosystems by industrial contamination, environmental monitoring is a standard procedure used for assessing the current status of an environment or trends in environmental parameters. Measurement of metal concentrations at different trophic levels followed by their statistical analysis using exploratory multivariate methods can provide meaningful information on the status of environmental quality. In this context, the present paper proposes a novel chemometric approach to standard statistical methods by combining the Block clustering with Partial least square (PLS) analysis to investigate the accumulation patterns of metals in anthropized terrestrial ecosystems. The present study focused on copper, zinc, manganese, iron, cobalt, cadmium, nickel, and lead transfer along a soil-plant-snai food chain, and the hepatopancreas of the Roman snail (Helix pomatia) was used as a biological end-point of metal accumulation. RESULTS: Block clustering deliniates between the areas exposed to industrial and vehicular contamination. The toxic metals have similar distributions in the nettle leaves and snail hepatopancreas. PLS analysis showed that (1) zinc and copper concentrations at the lower trophic levels are the most important latent factors that contribute to metal accumulation in land snails; (2) cadmium and lead are the main determinants of pollution pattern in areas exposed to industrial contamination; (3) at the sites located near roads lead is the most threatfull metal for terrestrial ecosystems. CONCLUSION: There were three major benefits by applying block clustering with PLS for processing the obtained data: firstly, it helped in grouping sites depending on the type of contamination. Secondly, it was valuable for identifying the latent factors that contribute the most to metal accumulation in land snails. Finally, it optimized the number and type of data that are best for monitoring the status of metallic contamination in terrestrial ecosystems exposed to different kinds of anthropic polution.

Assessing the effects of different pectins addition on color quality and antioxidant properties of blackberry jam.

BACKGROUND: In the last years pectin and other hydrocolloids were tested for improving the color stability and the retention of bioactive compounds in gelled fruit-based products. In line with these concerns, our study has been directed to quantify the changes in antioxidant status and color indices of blackberry jam obtained with different types of pectin (degree of esterification: DE, degree of amidation: DA) and doses in response to processing and storage for 1, 3 and 6 months at 20°C. RESULTS: Blackberry jam was obtained by a traditional procedure used in households or small-scale systems with different commercial pectins (HMP: high-methoxyl pectin, LMP: low-methoxyl pectin and LMAP: low-methoxyl amidated pectin) added to three concentrations (0.3, 0.7 and 1.0%) and investigated in terms of total monomeric anthocyanins (TMA), antioxidant capacity expressed as ferric reducing antioxidant power (FRAP), total phenolics (TP), color density (CD) and percent of polymeric color, PC (%). Thermal processing resulted in significant depreciation of analyzed parameters reported to the corresponding values of fresh fruit as follows: TMA (69-82%), TP (33-55%) and FRAP (18-52%). Biologically active compounds and color were best retained one day post-processing in jams with LMAP followed by samples with LMP and HMP. Storage for 6 months brings along additional dramatic losses reported to the values recorded one day post-processing as follows: TMA (31-56%), TP (29-51%) and FRAP (20-41%). Also, both processing and storage resulted in significant increases in PC (%). The pectin type and dosage are very influential factors for limiting the alterations occurring in response to processing and storage. The best color retention and the highest TMA, TP and FRAP were achieved by LMAP, followed by LMP and HMP. Additionally, a high level of bioactive compounds in jam could be related to a high dose of pectin. LMAP to a level of 1% is the most indicated to provide the highest antioxidant properties in jam. CONCLUSIONS: The retention of bioactive compounds and jam color stability were strongly dependent on the pectin type and dosage. By a proper selection of pectin type and dose could be limited the losses recorded in response to processing and storage.

The occurrence of mycotoxins in wheat from western Romania and histopathological impact as effect of feed intake.

BACKGROUND: The goal of this study has been to evaluate the extent of mycotoxins contamination and their co-occurrence in wheat grain intended for animal feed. A total of 52 wheat samples were collected from the harvest of two consecutive years (2010, 2011) from two counties (Timis and Arad) located in Western Romania and the presence of ochratoxin A (OTA), deoxynivalenol (DON), zearalenone (ZON) and fumonisins (FUMO) was determined by enzyme-linked immunosorbent assay (ELISA). In order to evaluate the toxicological impact of mycotoxins, naturally contaminated wheat samples were administered in rats feed for one month. RESULTS: The mycotoxin with the highest incidence in wheat samples was DON due to agro-climatic conditions typical for the west part of Romania. DON was found in 73.08% of samples harvested in 2010 and the highest level of contamination was 3390 ppb. The incidence of DON in sample from 2011 was lower than those of 2010, with a frequency of occurrence of 19.23%. The occurrence of ZON was in the range 69.23-76.92%, with an average value of 187.74 ppb. The OTA content in wheat was below the maximum tolerable limit established by EU Commission regulation for feed legislation (250 ppb). For FUMO the lowest percentage of positive samples was registered (15.38% in wheat sample harvested in 2010 and 11.54% positive samples in 2011). With respect to the co-occurrence of Fusarium mycotoxins, the results proved that ZON was found as a co-contaminant together with DON, especially when climatic conditions for development of fungus are favorable (high air humidity). The differences recorded between investigated localities and their classifications according to the mycotoxin kind and year of harvest were carried out by principal components analysis (PCA). The histopathological and immunohistochemical evaluation performed by hematoxylin and eosin (HE) staining technique as well as by assessing the vascular endothelial growth factor (VEGF) revealed significant modification of kidney, liver and spleen cells in the case of DON and FUMO. In terms of toxicity induced by OTA and ZON it was highlighted mixed normal and necrotic areas in liver, while histological assessment revealed normal VEGF expression in kidneys. CONCLUSIONS: Although none of the analyzed samples exceeding the stipulated maximum limits for cereals used as feed, a high incidence of mycotoxins produced by Fusarium species have been recorded (DON and ZON) in wheat samples harvested in Western Romania. Also, histopathological evaluation revealed significant tissue lesions in liver and kidney of rats after one month of feeding with natural contaminated wheat.

Use of enzymatic tools for biomonitoring inorganic pollution in aquatic sediments: a case study (Bor, Serbia).

BACKGROUND: Sediment bacterial communities are key players in biogeochemical cycling of elements in the aquatic environment. Copper mining, smelting, and processing operations located in Bor area (Serbia) are major environmental hot spots in the lower Danube Basin and Western Balkans. In the present study, we evaluate the influence of trace element (TE) concentration in sediments and physico-chemical properties of water on sediment microbial communities in water streams adjacent to the Copper Smelter Complex Bor (RTB Bor, Serbia). The degree to which metabolic activities of bacterial biota inhabiting differently polluted sites is inhibited by inorganic pollution were compared using selected enzymatic bioindicators. RESULTS: Cu, Zn, Pb, and As concentrations systematically exceeded the target values for metal loadings in aquatic sediments. Water electrical conductivity (WEC) followed the same pattern of spatial variation, irrespective of season. Interestingly, the most intense enzymatic activity occurred at the reference site although this site showed the greatest TE levels in aquatic sediments. Catalase activity (CA), potential dehydrogenase activity (PDA), actual dehydrogenase activity (ADA), urease activity (UA), and phosphatase activity (PA) in aquatic sediments displayed heterogeneous patterns of spatio-temporal variation. Inorganic pollution greatly affected CA, ADA, and PDA, but much less so UA and PA. Canonical correlation analysis showed that pH and WEC were the strongest determinants of enzymatic activity in bacterial biota, with the latter variable being reversely correlated with the enzymatic indicator of sediment quality (EISQ). The median values of EISQ increased with distance from the major sources of pollution. In addition, it was found that sites with different degrees of inorganic pollution can be appropriately classified by applying cluster analysis to EISQ, TE levels in sediments, and physico-chemical properties of water. CONCLUSIONS: Because EISQ can precisely identify changes in overall enzymatic activity of sediment bacterial communities, this enzymatic bioindicator has a great potential for biomonitoring the current status of inorganic pollution in aquatic ecosystems.

Bioaccumulative and conchological assessment of heavy metal transfer in a soil-plant-snail food chain.

BACKGROUND: Copper (Cu), zinc (Zn), cadmium (Cd), and lead (Pb) can pose serious threats to environmental health because they tend to bioaccumulate in terrestrial ecosystems. We investigated under field conditions the transfer of these heavy metals in a soil-plant-snail food chain in Banat area, Romania. The main goal of this paper was to assess the Roman snail (Helix pomatia) usefulness in environmental monitoring as bioindicator of heavy metal accumulation. Eight sampling sites, selected by different history of heavy metal (HM) exposure, were chosen to be sampled for soil, nettle leaves, and newly matured snails. This study also aimed to identify the putative effects of HM accumulation in the environment on phenotypic variability in selected shell features, which included shell height (SH), relative shell height (RSH), and whorl number (WN). RESULTS: Significantly higher amounts of HMs were accumulated in snail hepatopancreas and not in foot. Cu, Zn, and Cd have biomagnified in the snail body, particularly in the hepatopancreas. In contrast, Pb decreased when going up into the food chain. Zn, Cd, and Pb correlated highly with each other at all levels of the investigated food chain. Zn and Pb exhibited an effective soil-plant transfer, whereas in the snail body only foot Cu concentration was correlated with that in soil. There were significant differences among sampling sites for WN, SH, and RSH when compared with reference snails. WN was strongly correlated with Cd and Pb concentrations in nettle leaves but not with Cu and Zn. SH was independent of HM concentrations in soil, snail hepatopancreas, and foot. However, SH correlated negatively with nettle leaves concentrations for each HM except Cu. In contrast, RSH correlated significantly only with Pb concentration in hepatopancreas. CONCLUSIONS: The snail hepatopancreas accumulates high amounts of HMs, and therefore, this organ can function as a reliable biomarker for tracking HM bioavailability in soil. Long-term exposure to HMs via contaminated food might influence the variability of shell traits in snail populations. Therefore, our results highlight the Roman snail (Helix pomatia) potential to be used in environmental monitoring studies as bioindicator of HM pollution.

Heavy metals health risk assessment for population via consumption of vegetables grown in old mining area; a case study: Banat County, Romania.

BACKGROUND: The aim of this study is to measure the levels of heavy metals (Fe, Mn, Zn, Cu, Ni, Cd and Pb) found in common vegetables (parsley, carrot, onion, lettuce, cucumber and green beans) grown in contaminated mining areas compared with those grown in reference clear area and to determine their potential detrimental effects via calculation of the daily metal intake (DImetal) and Target Hazard Quotients (THQ) for normal daily consumption of these vegetables, for male and female gender. RESULTS: Compared with the reference in contaminated areas, soil and plant contents of all analyzed metals are higher, usually over normally content for Mn, Zn, Cu, Cd and Pb. Particularly, in soil, higher values than intervention threshold values (ITV) were found for Cu and Pb and higher than maximum allowable limits (MAL) for Zn, Cu, Cd and Pb for parsley roots and leaves, carrot roots, cabbage, lettuce and cucumber. DImetal and THQ values for male and female were calculated for each vegetable and metal and for which oral reference doses exist. The combined THQ values calculated are concerning in that they are usually below the safe level of THQ<1 for all vegetables grown in reference area. In contaminated Moldova Noua (M) area the combined THQ exceeded the safe level only for parsley roots, while in more contaminated Ruschita (R) area combined THQ exceeded the safe level for parsley and carrot roots, lettuce and cabbage. Cd and Pb, most toxic metals to humans, have an increasing prevalence in the combined THQ for leafy (cabbage and lettuce) and fruit vegetables (cucumber). In the root vegetables only Pb has an increasing prevalence in combined THQ values. In all areas female THQ is higher than male THQ. CONCLUSION: The results of this study regarding metal contents in soils, vegetables, DImetal and THQ suggest that the consumption of some vegetables (especially parsley, carrot and cabbage and less for lettuce, cucumber and green beans) is not free of risks in these areas. The complex THQ parameter use in health risk assessment of heavy metals provides a better image than using only a simple parameter (contents of metals in soils and vegetables).