Hemocyte-specific responses to the peroxidizing herbicide fomesafen in the pond snail Lymnaea stagnalis (Gastropoda, Pulmonata)

Responses of circulating hemocytes were studied in Lymnaea stagnalis exposed to 10, 30, 90, and 270 microg/L fomesafen for 24 and 504 h. Flow cytometry was used to quantify fomesafen-induced production of reactive oxygen species (ROS), phagocytic activity on Escherichia coli, and oxidative burst when hemocytes were challenged by E. coli or phorbol 12-myristate-13-acetate (PMA). Lysosomal membrane damage was assessed, using the neutral-red retention time (NRRT) assay. Exposure to fomesafen for 24 h resulted in increase in ROS levels and decreases in phagocytosis and the oxidative burst in PMA-stimulated hemocytes. After 504 h, intracellular levels of ROS returned to normal, but phagocytosis of E. coli was still inhibited and the associated oxidative burst significantly reduced. After both durations of exposure, decreases of NRRT indicated that lysosome membrane fragility increased with fomesafen concentration. Potential implications for the health and survival of the snails and consequences on populations are discussed.     

Effects of environmental concentrations of atrazine on hemocyte density and phagocytic activity in the pond snail Lymnaea stagnalis (Gastropoda, Pulmonata)

Immunotoxicological effects of environmentally relevant concentrations (10, 23, 50, 100 microg/l) of atrazine were studied in Lymnaea stagnalis. Individual hemolymph sampling was performed at 0, 24, 48, 72, 96, 168, 336, 504 and 672 h during exposure. Every atrazine concentration induced a significant increase in the mean number of circulating hemocytes, without any concentration-response relation. A peak (1.6-fold increase) of hemocyte density was observed after 96 h of exposure. After 504 h, the number of hemocytes remained higher only in the snails exposed to the two highest concentrations. Granulocytes contributed most to the increase in hemocyte density in herbicide-exposed snails. Both short- (24 and 96 h) and long-term (504 h) exposures resulted in significant inhibition of hemocyte phagocytic activity upon E. coli. Over the long-term, phagocytosis recovered for the two lowest concentrations. After 504 h of exposure, every herbicide level resulted in a significant reduction of reactive oxygen species production in E. coli-stimulated hemocytes, which was not observed for short-term exposures.     

Chronic toxicity of tributyltin to development and reproduction of the European freshwater snail Lymnaea stagnalis (L.)

Chronic toxicity, growth and reproduction were measured in the freshwater gastropod Lymnaea stagnalis exposed to waterborne bis(tri-n-butyltin) oxide (TBTO) over a range of four nominal concentrations (0-10microg TBTl(-1)). Egg development was completely inhibited at 10microg TBTl(-1), whilst abnormal embryonic development was observed at 1microg TBTl(-1). For the solvent control and the 0.01microg TBTl(-1)treatment group, normal development of L. stagnalis was observed. Survivorship of hatchlings was significantly reduced by TBT at 1microgl(-1) while inhibition of shell growth of L. stagnalis was also observed at this concentration. The data were used to determine intrinsic growth rates (r) using two theoretical approaches (the Euler-Lotka equation and a Leslie Matrix). Both approaches showed that survival, fecundity and population growth rate were reduced at 1microg TBTl(-1). Interestingly, at 0.01microg TBTl(-1) snails showed a higher fecundity and growth rate than in the solvent control. The TBT concentration at which the r would equal zero (ECr(0)) and the population NOEC (No Observed Effect Concentration) were estimated. The population NOEC was defined as either the lower 95% confidence or lower 95% pointwise percentile limit of the ECr(0). Values obtained using the two different approaches were similar and thus a geometric mean was calculated to obtain a final representative population NOEC value for L. stagnalis of 2745ng TBTl(-1). The present data together with chronic toxicity TBT data for freshwater organisms, obtained from peer-reviewed literature, were used to construct a species sensitivity distribution (SSD). A predicted no effect concentration was then derived from the SSD (hazardous concentration at 5%, i.e., HC5 or 95% protection level). This SSD was compared with the SSD derived from saltwater species datasets. The HC5 value for saltwater species (3.55ng TBTl(-1); lower confidence limit: 1.93ng TBTl(-1)) was significantly lower than that for freshwater species (30.13ng TBTl(-1); lower confidence limit: 9.23ng TBTl(-1)), indicating that saltwater species are probably more susceptible to TBT than their freshwater counterparts.     

Effects of the mixture of diquat and a nonylphenol polyethoxylate adjuvant on fecundity and progeny early performances of the pond snail Lymnaea stagnalis in laboratory bioassays and microcosms

Effects of the bipyridylium herbicide diquat and tank-mix adjuvant Agral((R))90 were investigated on various life history traits of the freshwater pulmonate snail Lymnaea stagnalis. Trait expression was measured in simple laboratory bioassays on small size groups of snails, and under more complex, indoor microcosm conditions, on larger groups of snails. Microcosms were provided with sediment, plants, and fish, thus allowing a more complex level of intra and inter-specific interactions to develop. Treatments were performed with substances alone or in mixture, at concentrations ranging from 4.4 to 222.2mugl(-1) for diquat, and from 10 to 500mugl(-1) for Agral 90, under a fixed ratio design. Adult growth was negatively affected by diquat and its mixture with Agral 90 both at the highest concentrations (222.2 and 500mugl(-1), respectively). Fecundity expressed differently in bioassays and microcosms, but no effect of the chemicals could be observed on this trait. Progeny development was impaired by 222.2mugl(-1) diquat and its mixture with 500mugl(-1) Agral 90, as reflected by longer development time and reduced hatching rate of clutches laid by the exposed animals, as compared to the controls. Hatching data suggested that diquat bioavailability was lower in microcosms than under bioassay conditions. Consistently, chemical analysis showed that diquat disappeared more rapidly from the water in microcosms than in bioassays. Moreover, the differential expression of several life history traits under bioassays and microcosms conditions was probably also influenced by the level of intraspecific interaction, which differed among the systems. When significant, the effect of diquat was attenuated by the presence of Agral 90, indicating antagonistic interaction between the two substances. Such a deviation from additivity was partly validated statistically.     

Effect of humic acid on water chemistry, bioavailability and toxicity of aluminium in the freshwater snail, Lymnaea stagnalis, at neutral pH

The influence of humic acid on the water chemistry of environmentally relevant concentrations of Al at neutral pH was studied, together with its effect on the bioavailability and toxicity of Al in Lymnaea stagnalis. Humic acid significantly reduced the loss of Al from the water and increased the fraction of filterable Al, although this was a relatively small fraction of total Al. Filterable Al concentration in the presence or absence of humic acid was independent of initial Al concentration. Humic acid only partly reduced toxicity, as observed by a reduction in behavioural suppression, and had no effect on the level of Al accumulated in tissues. These results suggest that humic acid maintains Al in a colloidal form that is bioavailable to L. stagnalis. However, these colloidal Al-humic acid species were less toxic since behavioural toxicity was reduced. Humic acid may play an important role in limiting the toxicity of Al to freshwater organisms.     

Molluscicidal activity of Ferula asafoetida, Syzygium aromaticum and Carum carvi and their active components against the snail Lymnaea acuminata

The molluscicidal activity of dried root latex powder of Ferula asafoetida, flower-bud powder of Syzygium aromaticum and seed powder of Carum carvi against the snail Lymnaea acuminata was studied. The molluscicidal activity of all the three plant products was found to be both time and concentration dependent. The toxicity of S. aromaticum flower-bud powder (96 h LC(50):51.98 mg/l) was more pronounced than that of root latex powder of F. asafoetida (96 h LC(50):82.71 mg/l) and seed powder of C. carvi (96 h LC(50):140.58 mg/l). Ethanol extract was more toxic than other organic extracts. The ethanol extract of S. aromaticum (24h LC(50):83.53 mg/l) was more effective than that of F. asafoetida (24h LC(50):132.31 mg/l) and C. carvi (24h LC(50):130.61 mg/l) in killing the test animals. The 96 h LC(50) of column purified fraction of seed powder of C. carvi was 5.40 mg/l whereas those of flower-bud powder of S. aromaticum and dried root latex powder of F. asafoetida were 7.87 and 9.67 mg/l, respectively. The product of F. asafoetida, S. aromaticum and C. carvi may be used as potent molluscicides.     

The effects of endosulfan on the great ramshorn snail Planorbarius corneus (Gastropoda, Pulmonata): a histopathological study

In this study the great ramshorn snail (Planorbarius corneus), one of the most abundant gastropod of Turkish limnic systems, was investigated to determine the histopathological effects of endosulfan on the digestive gland, foot and mantle under laboratory conditions. Samples were collected from small artificial pools in Karaot at Gelendost-Isparta (southwest of Turkey), where agricultural activities are widespread. The snails were exposed to two sublethal concentrations of endosulfan (0.4 and 0.8 mg/l) for periods of 10, 20 and 30 days. Fifteen snails were kept in 2.5 l glass jars containing dechlorinated tap water and exposed under semi-static test (daily exchange of test water). All the testing was carried out on adult specimens, and snails were maintained on a photothermal period with 16 light hours at 22 +/- 2 degrees C. The histopathological examinations revealed the following changes: amoebocytes infiltration, dilatation in hemolymphatic spaces between the tubules, degeneration of cells, abnormal lumen, necrosis of cells and atrophy in the connective tissue of digestive gland; desquamation of the epithelium cells, changes in the number of mucocytes and protein gland cells, lipid vacuolus and atrophy of the columnar muscle fibers of the foot and mantle tissues. Pycnotic state of cells was also seen in the mantle tissues. Endosulfan caused significant histopathological alterations in the digestive gland, foot and mantle tissues of the snail, irrespective of concentrations of the pesticide and its exposure periods. The results are discussed, particularly in comparison to those of other aquatic organisms.     

Effects of the antibacterial agents tiamulin,olanquindox and metronidazole and the anthelmintic ivermectin on the soil invertebrate species Folsomia fimetaria (Collembola) and Enchytraeus crypticus (Enchytraeidae)

Veterinary pharmaceutical products such as antibacterial agents and antiparasitics are widely used to control diseases and promote production in the agricultural sector. Exposure of non-target organisms are a likely result of using manure from treated live stocks or from dung dropped on the field by grazing animals. The aim of this study was to determine the toxic threshold levels of three antibacterial agents (tiamulin, olanquindox and metronidazole) and one anthelmintic (ivermectin) to two species of soil dwelling organisms (springtails and enchytraeids), that are often found in bio-solids such as manure or dung. The antibacterial agents were not toxic to adults and effects on reproduction occurred generally above concentrations normally found in soil or dung. The threshold values for toxicity (10% reduced reproduction or EC10 values) were in the range of 61-111 mg kg(-1) dry soil for springtails and 83-722 mg kg(-1) dry soil for enchytraeids. Ivermectin was significantly more toxic with EC10 values of 0.26 mg kg(-1) dry soil for the springtails and 14 mg kg(-1) dry soil for the enchytraeids. A comparison of these results with rough estimates of likely and worse case environmental concentrations indicates a potential risk of ivermectin to non-target species such as springtails and enchytraeids, whereas direct toxic effect of antibacterial agents is very unlikely to occur at environmental realistic concentrations. However, indirect effects of antibacterial agents driven through changes in the food web cannot be abolished at this stage.     

Effects of heavy metal exposure on the condition and health of adult great tits (Parus major)

We examined the possible effects of heavy metal exposure on the quality and health of adult great tits (Parus major) at four study sites along a pollution gradient near a non-ferrous smelter in Belgium. Tarsus length, wing length, body mass and condition of great tits were compared with respect to study site, age (first-year and older great tits), sex and season (birds caught in winter and during breeding). Tarsus length did not differ significantly among study sites. The wing length of great tits was larger at the study site furthest from the smelter, especially for older great tits. The length of the outermost tail feathers, however, did not differ significantly among study sites. We found no signs of loss of body mass or condition towards the pollution source. The body mass and condition was lowest for female great tits at the site furthest from the smelter, especially during winter. Haematocrit values did not differ significantly among sites. Overall, we found no clear significant effects of heavy metal pollution on morphological measurements and health parameters of great tits.     

Histopathological effects of copper and lithium in the ramshorn snail, Marisa cornuarietis (Gastropoda, Prosobranchia)

The aim of this study was to determine and quantify effects of copper and lithium in tissues of early juveniles of the ramshorn snail, Marisa cornuarietis. For this purpose, hatchlings of M. cornuarietis were exposed for 7 days to a range of five different sublethal concentrations of copper (5, 10, 25, 50, and 75 μg Cu²⁺ L⁻¹) and lithium (50, 100, 200, 1000, and 5000 μg Li⁺ L⁻¹). Both metals changed the tissue structure of epidermis, hepatopancreas, and gills, varying between slight and strong reactions, depending on the copper and lithium concentration. The histopathological changes included alterations in epithelial and mucous cells of the epidermis, swelling of hepatopancreatic digestive cells, alterations in the number of basophilic cells, abnormal apices of digestive cells, irregularly shaped cilia and changes in the amount of mucus in the gills. The most sensible organ in M. cornuarietis indicating Cu or Li pollution is the hepatopancreas (LOECs were 10 μg Cu²⁺ L⁻¹, or 200 μg Li⁺ L⁻¹). In epidermis, mantle and gills relevant effects occurred with higher LOECs (50 μg Cu²⁺ L⁻¹, or 1000 μg Li⁺ L⁻¹). Base on LOECs, our results indicated that histopathological endpoints are high sensitivity to copper and lithium compared to endpoints for embryonic developmental toxicity.     

Evaluation of HODE-15, FDE-15, CDE-15, and BDE-15 toxicity on adult and embryonic zebrafish (Danio rerio)

Diphenyl ether and its derivatives are widely used in the industry of spices, dyes, agrochemicals, and pharmaceuticals. Following the previous study, we selected 4,4′-dihydroxydiphenyl ether, 4,4′-difluorodiphenyl ether, 4,4′-dichlorodiphenyl ether, and 4,4′-dibromodiphenyl ether as research objects. The LC₅₀ (96 h) values for these compounds in adult zebrafish were determined with the acute test. Also, developmental toxicities of the four substances to zebrafish embryos were observed at 24, 48, 72, and 96 hpf. All the LC₅₀ (96 h) values of these compounds were between 1 and 10 mg/L, suggesting that they all had moderate toxicity to adult zebrafish. The embryonic test demonstrated that with increasing doses, 4,4′-dihydroxydiphenyl ether decreased the hatching rate, while 4,4′-difluorodiphenyl ether, 4,4′-dichlorodiphenyl ether, and 4,4′-dibromodiphenyl ether delayed the hatching time but had little effect on final hatchability at 96 hpf. All of these compounds inhibited larval growth, especially 4,4′-dihydroxydiphenyl ether. Exposure to these chemicals induced embryo yolk sac and pericardial edema. Spine deformation was visible in hatched larvae after 96 hpf 4,4′-dihydroxydiphenyl ether exposure, while tail curvature was observed for the halogenated compounds. The overall results indicated that 4,4′-dihydroxydiphenyl ether, 4,4′-difluorodiphenyl ether, 4,4′-dichlorodiphenyl ether, and 4,4′-dibromodiphenyl ether all had significant toxicity on adult and embryonic zebrafish.     

Speciation of five arsenic species (arsenite, arsenate, MMAAV, DMAAV and AsBet) in different kind of water by HPLC-ICP-MS

A method using Ion Chromatography hyphenated to an Inductively Coupled Plasma-Mass Spectrometer has been developed to accurately determine arsenite (As(III)), arsenate (As(V)), mono-methylarsonic acid (MMAA(V)), dimethylarsinic acid (DMAA(V)) and arsenobetaine (AsBet) in different water matrices. The developed method showed a high sensitivity with detection limits for each arsenic species close to 0.4pg injected. Arsenite and arsenate were the major species found in surface and well waters, but AsBet and DMAA(V) were found in some surface waters, which has never been reported before, while in some natural mineral waters located in volcanic region, the arsenic content exceeded the maximal admissible arsenic content by European legislation standards and the predominant form was As(V).     

Heavy metal accumulation and histomorphological alterations in Aphanius fasciatus (Pisces,Cyprinodontidae) from the Gulf of Gabes (Tunisia)

The present study illustrates an analysis of histological changes; cadmium (Cd), copper (Cu) and zinc (Zn) accumulation; and metallothionein (MT) levels in normal and deformed Mediterranean killifish, Aphanius fasciatus (Pisces, Cyprinodontidae), collected from unpolluted (S1) and polluted areas (S2) in the Gulf of Gabes in Tunisia. Metal determination in water and sediment showed that the concentrations were significantly higher (p?Histopathological investigations revealed greater changes in gills, kidney, liver, and bone tissues of fish from the polluted area than those recorded in fish from the reference area. In comparison to normal fish of the polluted area (S2), tissue alterations were more developed in deformed specimens of this site. A possible relationship between metallic pollution, incidence of spinal deformities, and histological changes in A. fasciatus in the polluted site was discussed.     

Imidacloprid induced alterations in oxidative stress,biochemical, genotoxic,and immunotoxic biomarkers in non-mammalian model organism Galleria mellonella L. (Lepidoptera: Pyralidae)

Imidacloprid (IMI), a neonicotinoid insecticide, is widely used to control pests in agriculture. We investigated the changes in antioxidant enzyme activities, lipid peroxidation levels, biochemical effects, genotoxic effect, and immunotoxic effect of sublethal doses (0.25, 0.50, 0.75, and 1.00?µg) of IMI at different time periods (24, 48, 72, and 96?h) on a model organism, Galleria mellonella L. The results indicated that there were dose-dependent increases in both antioxidant enzyme activities (SOD and CAT) and MDA levels. Protein content was not affected by IMI at 24th and 48th, whereas it was decreased by the highest dose of IMI (1.00?µg) at 72nd and 96th h. Lipid and carbohydrate contents were reduced with increasing doses of IMI. Micronucleus frequency significantly increased in all IMI doses. All IMI doses caused a significant decrease in THC at 24th, 48th, and 72nd h. Our results can help to illustrate the effects of IMI in target organisms and indirectly may aid to discover potential risk of it on nontarget organisms. Future studies, at molecular levels, will be helpful in understanding the mechanism of action of IMI on these biomarkers.     

Distribution and accumulation of elements (As, Cu, Fe, Hg, Mn, and Zn) in tissues of fish species from different trophic levels in the Danube River at the confluence with the Sava River (Serbia)

Pikeperch (Sander lucioperca), European catfish (Silurus glanis), common carp (Cyprinus carpio), and gobies (Neogobius gymnotrachelus, Neogobius melanostomus) were collected from the Danube River (Belgrade section), and samples of liver, muscle, or whole-body composites (in the case of gobies) were analyzed for As, Cu, Fe, Hg, Mn, and Zn with inductively coupled plasma optical spectrometry to find out if there was a correlation between accumulation of these elements in predatory and prey species, as well as in pairs of species with overlapping diets. Concentrations of all analyzed elements were either higher (Cu, Fe, Mn, Zn) in liver than in muscle, or equal (As, Hg), except for Hg in carp, which was higher in muscle. Mercury concentration in liver and muscle of predators (catfish, pikeperch) was significantly (<10^?4) higher than in prey fishes (carp and gobies). The results indicate that Hg concentration was biomagnified through the food chain. Concentrations of As, Fe, and Hg in carp liver and gobies whole-body composite were similar, but carp had significantly (<10^?4) higher values of Zn and Cu in liver. The regression analysis and trendline equations indicate that the concentrations of all tested elements, except for As in liver, and Mn and Fe in muscle, were similar in predatory fish (pikeperch and catfish), on one hand, and in prey fish (carp and gobies), on the other hand. Distinctly high Zn concentration in carp is very common in this species due to its physiology. Concentrations of Hg and Zn were higher than the maximum acceptable concentration due to the high pollution level in this section of the Danube River, accordingly posing a risk for the human consumption of these fish species.     

Comparing the response of biochemical indicators (biomarkers) and biological indices to diagnose the ecological impact of an oil spillage in a Mediterranean river (NE Catalunya, Spain)

Three biomarkers of hydrocarbon exposure, liver 7-ethoxyresourfin-O-deethylase activity (EROD), fluorescent hydrocarbon compounds (FACs) in bilis, and the liver antioxidant enzyme catalase (CAT) were examined in the autochthonous fish species Barbus meridionalis collected in the river Fluvià (NE Catalunya, Spain) after an oil spillage. Four different locations were sampled, including the impacted site, upstream and downstream sites and a reference site. Biomarker responses were compared with diatom and macroinvertebrate community assemblage metrics (Specific Pollution Sensitivity index - IPS, and Iberian Biological Monitoring Working Party - IBMWP, respectively). Chemical analyses denoted that polycyclic aromatic hydrocarbon levels in sediment were much higher at the impacted site than in downstream reaches. Four fold increase of EROD activity together with increased levels of biliary FACs in barbs collected at the spilled site indicated exposure of inhabiting fish to the oil. Additionally, CAT activity was significantly depressed (four fold) when compared to other stations, thus suggesting that fish collected from the impacted sites could be more susceptible to suffer oxidative stress. Biological indices (particularly that of the diatom community IPS) showed slight significant effects between control and impacted sites, indicating that more tolerant taxa were favoured because of the oil spillage. These results support the need to include biochemical responses measured in local species in monitoring programmes aimed to diagnose specific pollution effects in stressed river ecosystems.     

Persistent organochlorines in minke whale (Balaenoptera acutorostrata) and their prey species from the Antarctic and the North Pacific

Persistent organochlorines such as polychlorinated biphenyls (PCBs), DDTs, chlordane compounds (CHLs), hexachlorocyclohexanes (HCHs) and hexachlorobenzene (HCB) were determined in the blubber of minke whale and its diet collected from the Antarctic and the North Pacific Oceans. Residue levels of these compounds (except HCB) in minke whale from the Antarctic were apparently lower than those from the North Pacific. This is due to the lower levels of these pollutants in the southern hemisphere than in the northern hemisphere and the specific feeding habit of the minke whale from the Antarctic which feeds on lower trophic organisms, primarily euphausiids. The north-south difference for HCB residue levels was small, reflecting its dispersible nature through long-range atmospheric transport. Compositions of DDT and CHL compounds in minke whale from the Antarctic were similar to those from the North Pacific. However, the composition of HCH isomers was different between the Antarctic and the North Pacific as was observed in their diet, suggesting a larger or on-going usage of lindane in the southern hemisphere countries. In minke whale from the Antarctic, the elevated level of PCBs residues was noted during a period of 1984 to 1993, implying a continuous discharge of PCBs in the southern hemisphere. A similar discharge was also suggested in the North Pacific, while a decreasing contamination by DDTs was apparent.     

Exposure of Brassica juncea (L) to arsenic species in hydroponic medium: comparative analysis in accumulation and biochemical and transcriptional alterations

Arsenic (As) contamination in the environment has attracted considerable attention worldwide. The objective of the present study was to see the comparative effect of As species As(III) and As(V) on accumulation, biochemical responses, and gene expression analysis in Brassica juncea var. Pusa Jaganath (PJn). Hydroponically grown 14-day-old seedlings of B. juncea were treated with different concentrations of As(III) and As(V). Accumulation of total As increased with increasing concentration of both As species and exposure time, mainly in roots. Reduction in seed germination, root–shoot length, chlorophyll, and protein content were observed with increasing concentration and exposure time of both As species, being more in As(III)-treated leaves. PJn variety showed that antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX)) and stress-related parameters (cysteine, proline, and malondialdehyde (MDA)) were stimulated and allows plant to tolerate both As species. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis in leaves showed significant changes in protein profile with more stringent effect with As(III) stress. Semiquantitative RT-PCR analysis showed regulation in expression of phytochelatin synthase (PCS), metallothionine-2 (MT-2), glutathione reductase (GR), and glutathione synthetase (GS) genes under both As(III) and As(V) stresses. Results suggested that accumulation and inhibition on physiological parameters differ according to the As species, while molecular and biochemical parameters showed a combinatorial type of tolerance mechanism against As(III) and As(V) stresses.     

Nitrate causes deleterious effects on the behaviour and reproduction of the aquatic snail Potamopyrgus antipodarum (Hydrobiidae, Mollusca)

Nitrate (NO₃ ^?) is present in aquatic ecosystems as a natural component of the nitrogen cycle. However, in the last decades, several human activities are the causes of the rising amounts of organic matter and inorganic nitrogen nutrients in aquatic ecosystems, causing notable increase of nitrate above background natural levels. In spite of the toxicity of nitrate to aquatic animals, there are relatively few studies on the chronic toxicity of this compound to invertebrates. The aim of our study is to assess the effect of chronic (35 days) exposure to nitrate on the behaviour (velocity of movement) and reproduction (number of newborns) of the aquatic snail Potamopyrgus antipodarum. Four actual concentrations of nitrate were used (21.4, 44.9, 81.8 and 156.1 mg?N-NO₃ ^?/L). In each treatment, 12 animals were individually monitored for velocity (weekly) and newborn production (every 3–4 days). Velocity was recorded using quantitative video monitoring. Our results showed that nitrate did not cause mortality, but it reduced the velocity of movement (at 44.9, 81.8 and 156.1 mg?N-NO₃ ^?/L) and number of live newborns (in all tested concentrations). Reproductive impairment was caused at realistic nitrate concentrations which is relevant to the risk assessment of this compound. Our study contributes to the knowledge of the chronic effects of nitrate on the behaviour and reproduction of an aquatic snail.