Growth of the damselfly Ischnura heterosticta is better in saline water than freshwater

Increasing salinity has the potential to affect freshwater organisms. Yet sub-lethal effects of salinity on macroinvertebrates are poorly understood. Growth and development of Ischnura heterosticta (Odonata: Coenagrionidae) was experimentally shown to be faster in 5-20 mS/cm than 0.1-1 mS/cm, while in 35 mS/cm all individuals died. In 30 mS/cm about half died and growth was similar to the 0.1 mS/cm treatment. The salinity-growth relationship cannot be explained indirectly, that is salinity affecting the survival of their prey. Tissue content and concentration of Ca, Mg, Na and K in emerged adults showed no evidence of deficiencies at low salinity. Heart beat rate was similar across treatments, except at 35 mS/cm, where it was slower. Respiration and feeding were similar at 0.1, 10 and 30 mS/cm. While there are similarities in I. heterosticta and other species' salinity response, there are differences and studies on more species are urgently needed.     

Do laboratory salinity tolerances of freshwater animals correspond with their field salinity?

The degree to which laboratory derived measures of salinity tolerance reflect the field distributions of freshwater biota is uncertain. In this paper we compare laboratory-derived acute salinity tolerance (LC(50) values) of freshwater macroinvertebrates (range 5.5-76 mS/cm) and fish (range 2.7-82 mS/cm) from southeastern Australia with the salinity from which they have been collected in the field. Only 4% of the macroinvertebrates were collected at salinity levels substantially higher than their 72-h LC(50) obtained from directly transferring animals from low salinity water to the water they were tested (direct transfer LC(50)). This LC(50) value was correlated with the maximum salinity at which a species had been collected. For common macroinvertebrates, the maximum field salinity was approximated by the direct transfer 72-h LC(50). For adult freshwater fish, 21% of species were collected at salinities substantially greater than their acute direct transfer LC(50) and there was a weak relationship between these two variables. Although there was a weak correlation between the direct transfer LC(50) of early life stages of freshwater fish and the maximum field salinity, 58% of the field distribution were in higher than their LC(50) values. In contrast, LC(50) determined from experiments that acclimated adult fish to higher salinity (slow acclimation) provided a better indication of the field distribution: with only one fish species (7%) being in conflict with their maximum field salinity and a strong positive relationship between these variables. This study shows that laboratory measures of acute salinity tolerance can reflect the maximum salinity that macroinvertebrate and fish species inhabit and are consistent with some anecdotal observations from other studies.     

潜水丁坝在湖滨带生态恢复中的应用

为保护和稳固湖滨带,在太湖梅梁湾进行生态恢复研究,并成功地实施了示范工程.沿湖滨带共构筑14道潜水丁坝,每道长23～28 m,坝间距为40～80 m,坝上种植芦苇和菰.构筑的潜水丁坝能经受住太湖常见大风袭击和湖流淘蚀,芦苇和菰等长势良好,湖滨原有芦苇带在一定水深范围内每年向湖延伸2～5 cm,坝上其他水生植物和底栖动物螺蛳等自然繁衍.潜水丁坝所起作用为利用自然力改变岸边流场,使泥沙在湖滨带预定沉积区内沉积,减缓岸边侵蚀和降低沉积物再悬浮,从而为湖滨带水生植物生长创造良好的生境条件.研究发现,在微风条件下,潜水丁坝群能有效拦截麇集于岸边的蓝绿藻,并在湖滨带进行消化降解.研究提出的构筑潜水丁坝并种植水生植物的方案为湖滨带生态恢复提供了一种新的探索途径.     

Water Quality of Medium Size Watercourse Under Baseflow Conditions: The Case Study of River Sutla in Croatia

The study on medium size river Sutla in Croatia indicated considerable water contamination at specific sites during the baseflow period, probably associated to low flow-rate (0.73-68.8 m3 s(-1)), and consequently low dilution capacity of this river. Various aspects of contamination were observed: increased conductivity to 1,000 microS cm(-1), decreased dissolved oxygen level to 50%, 4-5 degrees C increased water temperature, increased concentrations of several dissolved trace elements (e.g., maximal values of Li: 45.4 microg l(-1); Rb: 10.4 microg l(-1); Mo: 20.1 microg l(-1); Cd: 0.31 microg l(-1); Sn: 30.2 microg l(-1); Sb: 11.8 microg l(-1); Pb: 1.18 microg l(-1); Ti: 1.03 microg l(-1); Mn: 261.1 microg l(-1); and Fe: 80.5 microg l(-1)) and macro elements (e.g., maximal values of Na: 107.5 mg l(-1); and K: 17.3 mg l(-1)), as well as moderate or even critical fecal (E. coli: 4,888 MPN/100 ml; total coliforms: 45,307 MPN/100 ml; enterococci: 1,303 MPN/100 ml) and organic pollution (heterotrophic bacteria: 94,000 cfu/ml). Although metal concentrations still have not exceeded the limits considered as hazardous for aquatic life or eventually for human health, the observed prominent increases of both metal concentrations and bacterial counts in the river water should be considered as a warning and incentive to protect the small and medium size rivers from the future deterioration, as recommended by EU Water Framework Directive.     

The effects of engineered nanoparticles on survival, reproduction, and behaviour of freshwater snail, Physa acuta (Draparnaud, 1805)

Increasing uses of engineered nanoparticles (ENPs) in commercial products and industrial applications has eventually resulted to their releases into atmospheric, terrestrial, and aquatic environments. However, knowledge gaps in ENPs toxicity, fate, and behaviour currently limit our ability to quantify risk assessment of materials with nanoscale dimensions, and therefore, the extent of the resultant environmental impacts remains unknown. In the present study, we evaluated the effects of γ-alumina, α-alumina, modified TiO(2) (M-TiO(2)), and commercial TiO(2) (C-TiO(2)) ENPs on the survival, behaviour, and early life stages of the freshwater snail Physa acuta (Draparnaud). The toxicity evaluation was carried out after spiking commercial sand with ENPs concentrations of 0.005, 0.05, or 0.5 gk g(-1). Our findings suggest that increases of γ-alumina and α-alumina concentrations at sub-lethal level concentrations caused significant reduction in the embryo growth rate and embryo hatchability. In addition, these ENPs induced observable developmental deformities of the embryos. In addition, toxicity evaluations using acute 96-h and chronic 28-d tests showed exposure duration may be a significant factor in ENPs-induced toxicity. Therefore, long-term exposure of aquatic organisms to ENPs - potentially can alter certain ecological populations at different trophic levels - and may compromise the entire aquatic ecological functionality. The percentage hatchlings in test chambers containing 0.5 gk g(-1) γ-alumina and α-alumina concentration was 50% less to those observed in the controls. Our results suggest the embryonic growth and hatchability tests are useful endpoints in chronic sediment toxicity tests for determining the toxic thresholds of ENPs in sediment environment. Although no snail mortalities were observed during the static 96-h test containing sediment spiked with different concentrations of M-TiO(2), C-TiO(2), γ-alumina and α-alumina - the antioxidant enzymatic assay results indicated a significant change in antioxidant levels which altered peroxidation at 0.05 or 0.5 gk g(-1)concentrations for both γ-alumina and α-alumina.     

Acute and chronic toxicity of veterinary antibiotics to Daphnia magna

The acute and chronic toxicity of nine antibiotics used both therapeutically and as growth promoters in intensive farming was investigated on the freshwater crustacean Daphnia magna. The effect of the antibiotics metronidazole (M), olaquindox (OL), oxolinic acid (OA), oxytetracycline (OTC), streptomycin (ST), sulfadiazine (SU), tetracycline (TC), tiamulin (TI) and tylosin (TY) was tested in accordance to the ISO (1989) and OECD (1996) standard procedures. The acute toxicities (48-h EC50 value, mg/l) in decreasing order were OA (4.6), TI (40), SU (221), ST (487), TY (680) and OTC (approximately 1000). NOECs were 340 mg/l for TC and 1000 mg/l for M and OL. Toxic effect on reproduction occurred generally at concentrations, which were one order of magnitude below the acute toxic levels. The chronic toxicity (EC50 values, mg/l) in the D. magna reproduction test in decreasing order were TI (5.4), SU (13.7), TC (44.8) and OTC (46.2). The NOECs (mg/l) obtained in the reproduction test with OA, ST, TY and M were 0.38 for OA, 32 for ST, 45 for TY and 250 for M. The observed toxicity of OA to D. magna indicates that this substance, which is a commonly used feed additive in fish farms, has a potential to cause adverse effects on the aquatic environment.     

How are macroinvertebrates of slow flowing lotic systems directly affected by suspended and deposited sediments?

The effects of suspended and deposited sediments on the macroinvertebrates are well documented in upland streams but not in slower flowing lowland rivers. Using species found in lowland lotic environments, we experimentally evaluate mechanisms for sediments to affect macroinvertebrates, and in one experiment whether salinity alters the effect of suspended sediments. Suspended kaolin clay reduced feeding of Ischnura heterosticta (Odonata: Coenagrionidae) at high turbidity (1000-1500 NTU) but had no effects on feeding of Hemianax papuensis (Odonata: Aeshnidae) and Micronecta australiensis (Hemiptera: Corixidae). In freshwater (0.1 mS/cm), survival of Ischnura aurora was poor in clear water, but improved with suspended kaolin. Growth and feeding of I. aurora were unaffected by suspended sediments and salinity. Burial (1-5 mm) of eggs with kaolin or sand reduced hatching in Physa acuta (Gastropoda: Physidae), Gyraulus tasmanica (Gastropoda: Planorbidae) and Chironomus cloacalis (Diptera: Chironomidae). Settling sediments may pose greater risk to lowland lotic invertebrates than suspended sediments.     

Effects of fluoxetine on the reproduction of two prosobranch mollusks: Potamopyrgus antipodarum and Valvata piscinalis

Fluoxetine is a widely used antidepressant, frequently found in aquatic ecosystems. We investigated its effects on two freshwater prosobranch gastropods: Valvata piscinalis (European valve snail) and Potamopyrgus antipodarum (New Zealand mudsnail), which have different reproductive modes. The fecundity of V. piscinalis (cumulate number of eggs at day 42) was not affected with an NOEC of 100 μg/L nominal concentration (69 μg/L measured concentration). The mudsnail P. antipodarum responded in a biphasic dose-effect curve at low concentrations. The cumulate number of neonates at day 42 had an LOEC of 100 μg/L (69 μg/L) and an NOEC of 33.3 μg/L (13 μg/L), whereas the embryos in the brood pouch at day 42 only showed an LOEC of 3.7 μg/L (1 μg/L). We also observed histological effects in P. antipodarum (gonadal thickness). Among the sexual steroids we measured only testosterone which varied, independent of reproduction. Moreover the use of two closely related species highlights the interspecific variability.     

Effects of landfill leachate effluent and bisphenol A on glutathione and glutathione-related enzymes in the gills and digestive glands of the freshwater snail Bellamya purificata

Environmental contaminants with estrogenic activity have recently attracted attention due to their potential detrimental effects on the reproduction of human and wildlife. The aim of this study was to evaluate the use of endogenous glutathione and glutathione-related enzymes as biomarkers of exposure to landfill leachate effluent and bisphenol A (BPA) in the freshwater snail, Bellamya purificata. Following exposure to 1%, 5% and 10% landfill leachate effluent and 1, 10, 50 and 100mugl(-1) BPA for 0, 2, 7 and 15d, activities of glutathione S-transferase (GST), selenium-dependent glutathione peroxidase (SeGPx) and glutathione reductase (GR) and levels of total glutathione were measured in the gills and digestive glands of the snails. GST and total glutathione were the most sensitive parameters in both exposure scenarios. GST activities increased by about 80%, while total glutathione decreased to 70% and 80% in the gills and digestive glands, respectively. In contrast, SeGPx and GR activities remained at the same levels in all the treatment groups compared with those of controls. The results indicated that among glutathione and glutathione-related enzymes, GST activity and total glutathione level, which showed dose-dependent dynamics, could be used as biomarkers of aquatic ecosystems contaminated with landfill leachate.     

An invertebrate embryo test with the apple snail Marisa cornuarietis to assess effects of potential developmental and endocrine disruptors

A novel invertebrate embryo test with the apple snail, Marisa cornuarietis, comprising a test protocol for the following developmental endpoints is described: formation of eyes and tentacles, heart rate, hatching, weight after hatching. To evaluate effects on embryonic development, the snails were treated in a first step with 250 or 500 microg/l cadmium. Sublethal effects in terms of a significant delay in hatching could be found in the 250 microg/l treated animals, whereas 500 microg/l Cd were lethal for the snail embryos. To test endocrine disrupting chemicals with this protocol, experiments with bisphenol A (50 microg/l, 100 microg/l) and 17alpha-ethinylestradiol (10 microg/l) were performed. In both treatments an increase of weight after hatching was observed as well as a significant decline in the heart rate of the embryos. As shown here, the sensitivity of M. cornuarietis embryos test is equal or even higher than other test species like zebrafish embryos and, therefore, this test can be regarded as an alternative or supplement for ecotoxicological studies.     

Toxicity of pirimiphos-methyl: I. The acute and subacute oral toxicity in albino rats

In an acute study, albino rats of both sexes were orally administered graded doses of Pirimiphosmethyl, and the statistically computed median lethal dose (LD-50) were 1861 and 1667 mg/kg body weight for male and female rats respectively. No treatment related changes were discernible with regard to food intake, growth, gross or histopathology of the organs. In a time-course study, the correlation between symptoms and degree of esterase inhibition was examined in rats administered the minimum lethal dose (MLD: 1000 mg/kg b.w.) of the insecticide. Time-course inhibition pattern of both cholinesterase (ChE) and non-specific carboxylesterase (NSE) activities in brain and plasma revealed maximum inhibition at 24 h post-treatment which correlated well with the intensity of symptoms. In a subacute study, groups of male rats were fed dietary Pirimiphos-methyl at 0, 10, 250, 500 and 1000 ppm for 28 days. Food consumption and growth rate were not affected throughout the experimental period. At necropsy after 28 days, no gross pathological changes were seen in any of the organs except a slight increase in liver weight at 1000 ppm. Though no statistical differences were observed in the levels of hepatic transaminases, a significant increase in serum transaminase was evident. Significant increase in the activities of hepatic ALP, beta-GLR and serum ALP were evident at 500 and 1000 ppm. Further, significant inhibition of plasma PChE was evident at 250, 500 and 1000 ppm while the degree of inhibition of brain AChE was significant only at the higher dosages. No histopathological alterations were observed in any of the organs.     

Effect of spatial variation on salinity tolerance of macroinvertebrates in Eastern Australia and implications for ecosystem protection trigger values

Salinisation of freshwater has been identified as a serious environmental issue in Australia and around the world. Protective concentrations (trigger values) for salinity can be used to manage salinity impacts, though require locally relevant salinity tolerance information. 72-h acute salinity tolerance values were determined for 102 macroinvertebrates collected from 11 locations in four biologically distinct freshwater bio-regions in Northeast Australia and compared with sensitivities observed in Southeast Australia. The salinity tolerance of individual taxa was consistent across Northeast Australia and between Northeast and Southeast Australia. However, two distinct communities were identified in Northeast Australia using distributions of the acute tolerance values and a calculated index of salinity sensitivity. Salinity trigger values should therefore be representative of local or regionally relevant communities and may be adequately calculated using sensitivity values from throughout Eastern Australia. The results presented provide a basis for assessing salinity risk and determining trigger values for salinity in freshwater ecosystems at local and regional scales in Eastern Australia.     

A chronic bioassay with the estuarine amphipod Corophium volutator: test method description and confounding factors

Methods of conducting a chronic sediment toxicity test with the estuarine amphipod Corophium volutator are described. They consist of a 49-day exposure, after which mortality, growth and reproduction are determined. Pilot experiments were used to optimize test design parameters such as temperature, duration, feeding and refreshing regimes, and effects of indigenous organisms. By way of further validation, the present study focused on the effects of four different parameters: oxygen saturation, salinity, ammonium and nitrite. These confounding factors might play an important role especially if the test is used for risk assessment of field-contaminated sediments. It is concluded that the present experimental design is well suited for chronic sediment exposures with C. volutator. The test can be performed at a broad range of salinity values, provided that controls are performed at the same salinity. Results further demonstrate that with the endpoints growth and reproduction this chronic test procedure is a factor 7-18 more sensitive to ammonium and nitrate than the standardized acute bioassay (endpoint mortality).     

Integral assessment of estrogenic potentials of sediment-associated samples

GOAL, SCOPE AND BACKGROUND: Exogenic endocrine-active substances are also called 'Endocrine Disrupting Chemicals' (EDC). They imitate or hinder the function of natural endogenic hormones or disturb the synthesis or the metabolism of hormones or of hormone receptors. The Enzyme-Linked Receptor Assay (ELRA) can detect estrogenic and anti-estrogenic effects at the level of receptor binding and is a useful tool for the integrative detection of contaminant effects. Although the test system has been used repeatedly in sediment assessments, the questions have remained concerning how it responds to variations in the physico-chemical matrix. For some bioassays, the salinity of the sample is a critical factor. This is especially relevant when testing wastewater samples or when sediment-associated samples in the tidal reaches of rivers are tested. Sediments in the tidal reaches of rivers change their salinity several times a day. Against this background, it would be beneficial to have a test procedure of known salinity tolerance. On account of this, the salinity tolerance of the ELRA was tested, assessed with reference substances at several salinity levels, and compared with the E-Screen method and a Yeast Estrogen Screen (YES), which are also frequently applied in environmental testing. The aim of this paper was to explore when the salinity limits within these test procedures are applicable. The trials should reveal the working range to be expected, characterize the salinity-dependent variations in sensitivity of the test, and provide options for methodological adjustments to improve the stability against increased salinity. METHODS: The ELRA was carried out with the human Estrogen Receptor alpha. (ER) using the same principle like a competitive immunoassay based on ligand-protein interaction. However, an essential difference is the use of a physiologically relevant receptor instead of an antibody as a linking protein. The ELRA measures the competition of sample estrogens and anti-estrogens against estradiol supplied as a BSA-coating conjugate for the binding site of dissolved ER. Estradiol or xeno-estrogen binding is quantified by a biotynilated anti-ER antibody and the subsequent measurement of peroxidase activity by a streptavidin-POD-biotin complex. The E-Screen was performed with the human breast cancer cell line MCF-7, which expresses the estrogen receptor constitutively. Cell proliferation depends on binding of estrogens or xeno-estrogens with the receptor. After incubation, estrogen-dependent cell growth was measured by sulforhodamin B staining. The YES was performed with a recombinant yeast strain, transfected with a receptor and a reporter plasmid bearing the estrogen receptor and a vitellogenin gene fused with the reporter gene lacZ. Estrogen or xeno-estrogen-dependent gene induction was measured indirectly by LacZ activity. The salinity levels were simulated in varying concentrations with NaCl from 0 to 40 per thousand or Artificial Sea Water (ASW) from 0 to 32 per thousand. RESULTS: The study characterized the factor 'salinity' for the prospective application fields of the ELRA. With reference substances such as 17-beta-estradiol, the ELRA showed classical sigmoidal concentration-effect relations in a range from 0.05 to 100 microg/l under physiological conditions. After a methodological adjustment to compensate decreasing receptor-binding affinity of estrogens and xeno-estrogens at higher salinity levels, the ELRA became applicable under salinity conditions up to concentrations of 20.5 per thousand. In tests, the ELRA reached under the influence of salinity a mean limit of detection of 0.062 microg/l 17-beta-estradiol. The mean relative inter-test error was around 11%. Above concentrations of 20.5 per thousand there is a risk of false negative assessment. Compared with the E-Screen method using the MCF7 cell line and the yeast estrogen test system (YES), the ELRA shows a lower sensitivity to 17-beta-estradiol. In the E-Screen, the cell proliferation was strongly reduced by sodium chloride induced cytotoxicity. In comparison with the E-Screen, the salinity tolerance of the YES and YAS methods is significantly higher. DISCUSSION: Despite adaption, total salinity tolerance could not be achieved with the ELRA. Freshwater samples were generally appraisable. Higher salinity levels above 20.5 per thousand would tend towards false negative results. The low inter-test error of 11% makes the ELRA suitable for the detection of estrogenic and anti-estrogenic potentials of single substances, substance mixtures, and of environmental samples. CONCLUSIONS: The ELRA is very fast and reproducible, it can be used for high-throughput screening in a microplate format at low cost, it is robust to microbial contamination, and is less susceptible to cytotoxic interferences than cell culture methods. RECOMMENDATIONS AND PERSPECTIVES: In their established form, the YES and the E-Screen methods are not applicable for liquid phase testing at higher salinity conditions. The salinity-adapted test version of the ELRA described here shows a broader working range for samples. Native water samples of more or less brackish origin or high-salinity effluent samples are testable. Results of tests with sediment associated samples of different salinity will be subject of a forthcoming publication.     

Shallow groundwater quality on dairy farms with irrigated forage crops

California's dairies are the largest confined animal industry in the state. A major portion of these dairies, which have an average herd size of nearly 1000 animal units, are located in low-relief valleys and basins. Large amounts of liquid manure are generated and stored in these dairies. In the semi-arid climate, liquid manure is frequently applied via flood or furrow irrigation to forage crops that are grown almost year-round. Little is known about the impact of manure management practices on water quality of the extensive alluvial aquifers underlying these basins. The objective of this work is to assess nitrate and salt leaching to shallow groundwater in a relatively vulnerable hydrogeologic region and to quantify the impact from individual sources on dairies. The complex array of potential point and nonpoint sources was divided into three major source areas representing farm management units: (1) manure water lagoons (ponds); (2) feedlot or exercise yard, dry manure, and feed storage areas (corrals); and (3) manure irrigated forage fields (fields). An extensive shallow groundwater-monitoring network (44 wells) was installed in five representative dairy operations in the northeastern San Joaquin Valley, CA. Water quality (electrical conductivity, nitrate-nitrogen, total Kjehldahl nitrogen) was observed over a 4-year period. Nitrate-N, reduced nitrogen and electrical conductivity (EC, salinity) were subject to large spatial and temporal variability. The range of observed nitrate-N and salinity levels was similar on all five dairies. Average shallow groundwater nitrate-N concentrations within the dairies were 64 mg/l compared to 24 mg/l in shallow wells immediately upgradient of these dairies. Average EC levels were 1.9 mS/cm within the dairies and 0.8 mS/cm immediately upgradient. Within the dairies, nitrate-N levels did not significantly vary across dairy management units. However, EC levels were significantly higher in corral and pond areas (2.3 mS/cm) than in field areas (1.6 mS/cm) indicating leaching from those management units. Pond leaching was further inferred from the presence of reduced nitrogen in three of four wells located immediately downgradient of pond berms. The estimated minimum average annual groundwater nitrate-N and salt loading from manure-treated forage fields were 280 and 4300 kg/ha, respectively. Leaching rates for ponds are estimated to be on the order of 0.8 m/year, at least locally. Since manure-treated fields represent by far the largest land area of the dairy, proper nutrient management will be a key to protecting groundwater quality in dairy regions overlying alluvial aquifers.     

Nitrate toxicity to aquatic animals: a review with new data for freshwater invertebrates

Published data on nitrate (NO3-) toxicity to freshwater and marine animals are reviewed. New data on nitrate toxicity to the freshwater invertebrates Eulimnogammarus toletanus, Echinogammarus echinosetosus and Hydropsyche exocellata are also presented. The main toxic action of nitrate is due to the conversion of oxygen-carrying pigments to forms that are incapable of carrying oxygen. Nitrate toxicity to aquatic animals increases with increasing nitrate concentrations and exposure times. In contrast, nitrate toxicity may decrease with increasing body size, water salinity, and environmental adaptation. Freshwater animals appear to be more sensitive to nitrate than marine animals. A nitrate concentration of 10 mg NO3-N/l (USA federal maximum level for drinking water) can adversely affect, at least during long-term exposures, freshwater invertebrates (E. toletanus, E. echinosetosus, Cheumatopsyche pettiti, Hydropsyche occidentalis), fishes (Oncorhynchus mykiss, Oncorhynchus tshawytscha, Salmo clarki), and amphibians (Pseudacris triseriata, Rana pipiens, Rana temporaria, Bufo bufo). Safe levels below this nitrate concentration are recommended to protect sensitive freshwater animals from nitrate pollution. Furthermore, a maximum level of 2 mg NO3-N/l would be appropriate for protecting the most sensitive freshwater species. In the case of marine animals, a maximum level of 20 mg NO3-N/l may in general be acceptable. However, early developmental stages of some marine invertebrates, that are well adapted to low nitrate concentrations, may be so susceptible to nitrate as sensitive freshwater invertebrates.     

Effects of the organophosphate insecticide azinphos-methyl on the reproduction and cholinesterase activity of Biomphalaria glabrata

Azinphos-methyl is an organophosphate insecticide used for pest control on a number of food crops in many parts of the world. The snail Biomphalaria glabrata is a freshwater gastropod widely distributed in South America, Central America and Africa. The aim of the present work was to investigate whether azinphos-methyl causes alterations in the reproduction of B. glabrata. To this end, gastropod pigmented specimens were exposed to various concentrations of the insecticide (0.021, 0.5, 2.5, and 5 mg L⁻¹) for either 2 or 14 d. Along 14 d, several reproduction parameters and cholinesterase (ChE) activity were evaluated. In each group, the number of egg masses, the number of eggs per mass, the number of hatchings, the time to hatching, and the survival of the offspring after one month of treatment was evaluated. The results showed that, depending on the concentration and time of exposure, azinphos-methyl induced alterations in the reproduction of B. glabrata. These alterations were mainly represented by a decrease in the number of egg masses, and, in some cases, by a lower number or even the total absence of hatchings. Thus, the gastropods exposed to 2.5 and 5 mg L⁻¹ of azinphos-methyl for 14 d showed ChE inhibitions higher than 35% along time and completely lost their ability to reproduce. On the other hand, exposure to high acute concentrations or exposure to low concentrations for 14 d resulted in ChE inhibition equal to or lower than 35% between 7 and 14 d of treatment and similar alterations in reproduction. These were represented by a decrease in the number of egg masses. At low pestice levels, the number of egg masses and the number of offspring resulted to be more sensitive biomarkers than ChE inhibition. It is concluded that the insecticide azinphos-methyl can cause a decline in the reproductive performance of B. glabrata.     

Reproduction of the estuarine and marine amphipod Corophium volutator (Pallas) in laboratory for toxicity testing

The acute whole sediment bioassay with the estuarine and marine amphipod Corophium volutator (Pallas) is broadly used within Europe. Hitherto, the test is carried out with field-collected animals. In order to provide a more standardized and continuously available test organism reproduction and growth experiments were performed for a period of one year in laboratory under simulated summer conditions (light/dark 16:8 at 15, 19, and 23 degrees C). C. volutator was the first time reproduced successfully in laboratory for several generations and independent from its natural life cycle also in winter. The females produced two to three broods with a mean number of 96 offspring. A mean growth rate of 0.07 mm per day was determined at 15 degrees C. The reproduction and growth experiments provide consequently the essential base for the development of a chronic toxicity test with C. volutator.     

Effects of pharmaceuticals on Daphnia survival, growth, and reproduction

Pharmaceuticals have been globally detected in surface waters, and the ecological impacts of these biologically-active, ubiquitous chemicals are largely unknown. To evaluate the aquatic toxicity of individual pharmaceuticals and mixtures, we performed single species laboratory toxicity tests with Daphnia magna, a common freshwater zooplankton. We conducted acute (6-day) and chronic (30-day) exposure pharmaceutical bioassays and evaluated survivorship and morphology of adults and neonates, adult length, resting egg production, brood size (fecundity), and the proportion of male broods produced (sex ratio). In general, exposure to a single pharmaceutical in the 1-100 microg/l range yielded no apparent effects on the normal life processes of Daphnia. However, chronic fluoxetine exposure (36 microg/l) significantly increased Daphnia fecundity, and acute clofibric acid exposure (10 microg/l) significantly increased sex ratio. A mixture of fluoxetine (36 microg/l) and clofibric acid (100 microg/l) caused significant mortality; the same fluoxetine concentration mixed with 10 microg/l clofibric acid resulted in significant deformities, including malformed carapaces and swimming setae. Mixtures of three to five antibiotics (total antibiotic concentration 30-500 microg/l) elicited changes in Daphnia sex ratio. We conclude: (1) individual and mixtures of pharmaceuticals affect normal development and reproduction of Daphnia magna, (2) aquatic toxicity of pharmaceutical mixtures can be unpredictable and complex compared to individual pharmaceutical effects, and (3) timing and duration of pharmaceutical exposure influence aquatic toxicity.     

Role of exogenous and endogenous silicon in ameliorating behavioural responses to aluminium in a freshwater snail

Aluminium accumulation by the freshwater snail Lymnaea stagnalis is correlated with behavioural depression which is ameliorated by addition of orthosilicic acid. We hypothesised that Si is relocated to the digestive gland in response to Al, leading to the formation of non-toxic hydroxyaluminosilicates (HAS). Exposure to 500 microg l(-1) Al for 30 days was associated with an initial period of behavioural depression, followed by apparent tolerance and subsequent depression, suggesting saturation of the cellular detoxification pathway during prolonged exposure. Exogenous Si (7.77 mg l(-1)) completely ameliorated all behavioural effects of Al but did not prevent its accumulation. In the presence of added Al, significantly more of this Si was accumulated by the tissues, compared to controls and snails exposed to Si alone. In snails exposed to Al plus Si, Al and Si concentrations were significantly correlated, with a ratio around 3:1 Al:Si, consistent with the presence of the non-toxic HAS protoimogolite.