The functional and physiological status of Gammarus fossarum (Crustacea; Amphipoda) exposed to secondary treated wastewater

Climate change scenarios predict lower flow rates during summer that may lead to higher proportions of wastewater in small and medium sized streams. Moreover, micropollutants (e.g. pharmaceuticals and other contaminants) continuously enter aquatic environments via treated wastewater. However, there is a paucity of knowledge, whether extended exposure to secondary treated wastewater disrupts important ecosystem functions, e.g. leaf breakdown. Therefore, the amphipod shredder Gammarus fossarum was exposed to natural stream water (n = 34) and secondary treated wastewater (n = 32) for four weeks in a semi-static test system under laboratory conditions. G. fossarum exposed to wastewater showed significant reductions in feeding rate (25%), absolute consumption (35%), food assimilation (50%), dry weight (18%) and lipid content (22%). Thus, high proportions of wastewater in the stream flow may affect both the breakdown rates of leaf material and thus the availability of energy for the aquatic food web as well as the energy budget of G. fossarum.     

Unexpected toxic interactions in the freshwater amphipod Gammarus pulex (L.) exposed to binary copper and nickel mixtures

To document the toxicity of copper and nickel in binary mixtures, freshwater amphipods Gammarus pulex were exposed to the metals given independently or as mixtures. Toxicity to Cu alone was relatively high: 96-h LC₁₀ and LC₅₀ were found at 91 and 196 μg L^?1, respectively. Toxicity to Ni alone was very low, with 96-h LC₁₀ and LC₅₀ of 44,900 and 79,200 μg L^?1, respectively. Mixture toxicities were calculated from single toxicity data using conventional models. Modeled toxicity was then compared with the measured toxicity of the binary mixture. Two kinds of mixtures were tested. Type I mixtures were designed as combinations of Cu and Ni given at the same effect concentrations, when taken independently, to identify possible interactions between copper and nickel. In type II mixtures, Cu concentrations varied from 0 to 600 μg L^?1 while the nickel concentration was kept constant at 500 μg L^?1 to mimic conditions of industrial wastewater discharges. Ni and Cu showed synergic effects in type I mixtures while type II mixtures revealed antagonistic effects. Low doses of Ni reduced Cu toxicity towards G. pulex. These results show that even for simple binary mixtures of contaminants with known chemistry and toxicity, unexpected interactions between the contaminants may occur. This reduces the reliability of conventional additivity models.     

Behavioural and feeding responses of Echinogammarus meridionalis (Crustacea, Amphipoda) to acid mine drainage

As a result of mining activities the exposure of metal sulphides to oxidation takes place with consequent release of acid mine drainage (AMD). Biomonitoring instruments have been proven to have the best deterrent effect upon polluters. A new approach in online biomonitoring, with the Multispecies Freshwater Biomonitortrade mark (MFB), was developed combining behavioural and feeding responses of the Portuguese indigenous benthic shredder, Echinogammarus meridionalis (Pinkster, 1973) (Crustacea, Amphipoda). These endpoints, along with mortality, were measured and analyzed for a gradient of sublethal doses of AMD. Original river water was used as the control and three doses of treatments were attained by adding increasing volumes of AMD to the control. The increase in AMD concentration and concomitant decrease in pH and increase in the concentration of most metals was followed by an overall increase of the mortality, decrease of locomotion/feeding activity and inhibition of the feeding rate. Mortality was observed in the two highest concentrations of AMD. Significant decrease in average locomotion/feeding activity took place in the second treatment. Although an inhibition of feeding was observed along the gradient of AMD concentration only in the highest concentration the feeding rate was significantly reduced.     

Comparison of arsenate and cadmium toxicity in a freshwater amphipod (Gammarus pulex)

Cadmium is largely documented on freshwater organisms while arsenic, especially arsenate, is rarely studied. The kinetic of the LC50s values for both metals was realized on Gammarus pulex. Physiological [i.e. metal concentration in body tissues, bioconcentration factor (BCF)] effects and behavioural responses (via pleopods beats) were investigated after 240-h exposure. Arsenate LC50 value was 100 fold higher than Cd-LC50 value after 240-h exposure, while concentrations in gammarids were similar for both metals at their respective LC50s. BCF decreased with increasing cadmium concentration while BCF remained stable with increasing arsenate concentration. Moreover, BCF was between 148 and 344 times lower for arsenate than cadmium. A significant hypoventilation was observed for cadmium concentrations exceeding or close to the 240h-LC50(Cd), while gammarids hyperventilated for the lowest arsenate concentrations and hypoventilated for the highest arsenate concentrations. We discussed the relationships between potential action mechanisms of these two metals and observed results.     

Suitability of Crangonyx pseudogracilis (Crustacea: Amphipoda) as an Early Warning Indicator in the Multispecies Freshwater Biomonitor (9 pp)

Background Biological monitors are increasingly important in 'Biological Early Warning Systems' (BEWS) for monitoring water quality. This study examines the freshwater amphipod Crangonyx pseudogracilis as a potential new indicator species when used in the Multispecies Freshwater Biomonitor (MFB). The MFB is an online continuous biomonitor which uses impedance conversion to record behavioural responses of vertebrates and invertebrates. Methods Four experiments were undertaken to establish: (1) if the electrical field generated by the MFB affected the organisms' behaviour, (2) if defined behaviours and their response to a gradient of ammonium chloride could be detected by the MFB, (3) if there was variation in the behaviour of C. pseudogracilis over a diel cycle, and (4) if behaviour changed significantly in response to a pulse of ammonium chloride. Results and Discussion Results showed no significant effect of the MFB's current on behaviour of C. pseudogracilis. Four behaviours; swimming, walking, grooming and inactivity, were observed and identified in the MFB. In the MFB, each behaviour changed significantly in response to an increasing gradient of ammonium chloride exposure. The MFB also detected increases in nocturnal activity by C. pseudogracilis. The MFB also detected a significant increase in activity after a pulse of ammonium chloride. Conclusion The range of behaviours exhibited by Crangonyx pseudogracilis, together with its ease of culture, suggest future potential of this species as an indicator species for the Multispecies Freshwater Biomonitor. Recommendations and Perspective . Further testing is required over a range of toxicants and concentration gradients to establish threshold responses and the full compliment of behaviours that could be useful in online biomonitoring.     

Enhanced metal and metalloid concentrations in the gut system comparing to remaining tissues of Gammarus pulex L

Invertebrate shredders such as Gammarus pulex are key species in contaminated stream ecosystems. Although a number of previous studies examining differences in metal accumulation between the gut system and remaining tissues of invertebrates exist, few focus on wide range of metals and metalloids that are relevant to contaminated systems. This study compared accumulation of the commonest (at study site) 15 metals and metalloids between the gut system including feces and remaining tissues of G. pulex. All metals and metalloids measured were significantly higher (p<0.001, except Cu p<0.005) in the gut system including feces than remaining tissues of G. pulex. Metals and metalloids in body tissues without the gut system including feces were significantly lower (Al, Cr, Fe and Mn (p<0.005), Sr and U (p<0.01), Co (p<0.05)) in content for a number of elements when compared to washed, whole G. pulex specimens. As well, all elements measured were significantly higher (all elements (p<0.005) except Cu and Co (p<0.05)) in gut system including feces than washed, whole G. pulex specimens. These results indicate that in G. pulex the uptake of all 15 metals and metalloids examined across the gut epithelium is minimalized or that sequestration of these elements in gut epithelial cells may occur.     

Effect of gender on physiological and behavioural responses of Gammarus roeseli (Crustacea Amphipoda) to salinity and temperature

The importance of potentially interacting factors in organisms responses to a stress are often ignored or underestimated in ecotoxicology. In laboratory experiments we investigated how gender, temperature and age influence the behaviour and the physiology of the freshwater amphipod Gammarus roeseli under salinity stress. Our results revealed a significant higher sensitivity of females in survival, ventilation and ionoregulation whereas no inter-age differences were reported. Water temperature also exerted a significant effect in survival and ventilation of G. roeseli. Some of those factors appeared to interact significantly. This study provides evidence that gender can affect organisms responses to a stressor and consequently has to be considered while assessing a stress impact. We discussed the potential relationships between biological and behavioural responses.     

Relationship between heavy metals pollution and genetic diversity in Mediterranean populations of the sandhopper Talitrus saltator (Montagu) (Crustacea, Amphipoda)

Trace metals are one of the groups of pollutants that reduce genetic variability in natural populations, causing the phenomenon known as “genetic erosion”. In this study we evaluate the relationship between trace metals contamination (Hg, Cd and Cu) and genetic variability, assessed using fluorescent Inter-Simple Sequence Repeats (fISSRs). We used eight populations of a well-established biomonitor of trace metals on sandy beaches: the amphipod Talitrus saltator. The trace metals analysis confirmed the ability of sandhoppers to accumulate Hg, Cd and Cu. Moreover, populations from sites with high Hg availability had the lowest values of genetic diversity. Our results validate the use of fISSR markers in genetic studies in sandhoppers and support the “genetic erosion” hypothesis by showing the negative influence of Hg contamination on sandhopper genetic diversity. Therefore, genetic variability assessed with fISSR markers could be successfully employed as a biomarker of Hg exposure.     

Validation of estuarine gammarid collectives (Amphipoda: Crustacea) as biomonitors for cadmium in semi-controlled toxicokinetic flow-through experiments

Concentrations of chemicals in organisms are regarded as being indicators of the bioavailable fraction of those substances in the environment. Three gammarid collectives (assemblages) from the Weser-estuary (NW Germany), showing different species compositions, were used as experimental units to evaluate their suitability as biomonitors. Uptake and clearance of cadmium was investigated in semi-controlled dynamic flow-through tests. Kinetic data obtained from two compartment models confirmed a net accumulation strategy of gammarids for Cd and resulted in similar bioconcentration factors (BCFs) within the same experimental treatments. Deviations in bioconcentration factors between two treatments (BCFs: 377-542 vs 947-1190, based on dry wt) obtained under similar exposures (17 +/- 2 and 23 +/- 2 microg Cd litre(-1)) could not be assigned to significant fluctuations of both Cd levels and particulate matter in the flow-through systems. The biological half-life of Cd in gammarids was 6 to 12 days. It is concluded that gammarid collectives of different species composition may be used as experimental units in biomonitoring studies. This conclusion was confirmed by homogeneous Cd concentrations in field samples of gammarids from the Weser-estuary (0.34 +/- 0.03 mg kg(-1) dry wt., mean +/- 95% confidence intervals, N = 58).     

Toxicity of imidacloprid to the terrestrial isopod Porcellio scaber (Isopoda, Crustacea)

Imidacloprid is a neonicotinoid insecticide with neurotoxic action that, as a possible alternative for commonly used organophosphorus pesticides, has gained registration in about 120 countries for use in over 140 agricultural crops. Only few data are available on its toxicity for soil invertebrates. We therefore assessed the effects of imidacloprid on survival, weight gain, feeding rate, total protein content, glutathione S-transferase activity (GST), and digestive gland epithelial thickness in juveniles and adults of the terrestrial isopod Porcellio scaber. After two weeks of feeding on imidacloprid-dosed food, weight gain (NOEC 5 microg/g dry food) and feeding rate (NOEC 10 microg/g) in juveniles, and feeding rate (NOEC<10 microg/g) and digestive gland epithelial thickness (NOEC<10 microg/g) in adults were most affected. In juveniles induction of GST activity and increase of total protein content per wet animal weight was detected at 5 microg/g dry food, whereas in adults a reduction of GST was observed at 25 microg/g (NOEC 10 microg/g). An estimate of actual intake rates suggests that imidacloprid affects isopods at similar exposure concentrations as insects. The toxicity of imidacloprid was similar to that of the organophosphorus pesticide diazinon, tested earlier using the same methods [Stanek, K., Drobne, D., Trebse, P., 2006. Linkage of biomarkers along levels of biological complexity in juvenile and adult diazinon fed terrestrial isopod (Porcellio scaber, Isopoda, Crustacea). Chemosphere 64, 1745-1752]. At actual environmental concentrations, diazinon poses a higher risk to P. scaber. Due to its increasing use in crop protection and higher persistence in soil, imidacloprid might however, be potentially more dangerous after long-term application. We conclude that toxicity testing with P. scaber provides relevant, repeatable, reproducible and comparable toxicity data that is useful for the risk assessment of pesticides in the terrestrial environment.     

Temporal and geographical genetic variation in the amphipod Melita plumulosa (Crustacea: Melitidae): Link of a localized change in haplotype frequencies to a chemical spill

Anthropogenic effects such as contamination affect the genetic structure of populations. This study examined the temporal and geographical patterns of genetic diversity among populations of the benthic crustacean amphipod Melita plumulosa in the Parramatta River (Sydney, Australia), following an industrial chemical spill. The spill of an acrylate/methacrylate co-polymer in naphtha solvent occurred in July 2006. M. plumulosa were sampled temporally between December 2006 and November 2009 and spatially in November 2009. Genetic variation was examined at the mitochondrial cytochrome c oxidase subunit I locus. Notably, nucleotide diversity was low and Tajima’s D was significantly negative amongst amphipods collected immediately downstream from the spill for 10 months. We hypothesize that the spill had a significant localized effect on the genetic diversity of M. plumulosa. Alternate explanations include an alternate and unknown toxicant or a localized sampling bias. Future proposed studies will dissect these alternatives.     

Acute toxicity tests with Daphnia magna, Americamysis bahia, Chironomus riparius and Gammarus pulex and implications of new EU requirements for the aquatic effect assessment of insecticides

Threshold concentrations for treatment related effects of 31 insecticides, as derived from aquatic micro-/mesocosm tests, were used to calibrate the predictive value of the European Tier-1 acute effect assessment on basis of laboratory toxicity tests with Daphnia magna, Chironomus spp., Americamysis bahia and Gammarus pulex. The acute Tier-1 effect assessment on basis of Daphnia (EC(50)/100) overall was protective for organophosphates, carbamates and most pyrethroids but not for neonicotinoids and the majority of insect growth regulators (IGRs) in the database. By including the 28-day water-spiked Chironomus riparius test, the effect assessment improves but selecting the lowest value on basis of the 48-h Daphnia test (EC50/100) and the 28-day Chironomus test (NOEC/10) is not fully protective for 4 out of 23 insecticide cases. An assessment on basis of G. pulex (EC(50)/100) is sufficiently protective for 15 out of 19 insecticide cases. The Tier-1 procedure on basis of acute toxicity data (EC(50)/100) for the combination of Daphnia and A. bahia and/or Chironomus (new EU dossier requirements currently under discussion) overall is protective to pulsed insecticide exposures in micro-/mesocosms. For IGRs that affect moulting, the effect assessment on basis of the 48-h Chironomus test (EC(50)/100) may not always be protective enough to replace that of the water-spiked 28-day C. riparius test (NOEC/10) because of latency of effects.     

Stressor identification and health assessment of fish exposed to wastewater effluents in Miho Stream, South Korea

This study evaluated the effects of an industrial wastewater treatment plant (IWTP) and a municipal wastewater treatment plant (MWTP) effluents on a variety of bioindicators ranging from biochemical, organism, and population-level responses in pale chub (Zacco platypus) and fish community structure. The Index of Biotic Integrity (IBI) indicated that the site upstream of these wastewater treatment plant discharges is in fair–good condition and downstream of the plant is in poor condition. The EROD (ethoxyresorufin-O-deethylase) activity, condition factor, and liver somatic index were significantly increased at the downstream site compared to those of the upstream site. The most significant change observed in pale chub population in the downstream site of the Miho Stream, relative to the upstream population, was the total absence of an younger age group. Stressors impacting the downstream site were identified as mostly organic or nutrient enrichment and habitat degradation associated with wastewater treatment plants. The results of causal analysis suggest that the primary causes affecting fish population in the downstream site are through both size-selective mortality caused by ammonia toxicity and recruitment failure caused by habitat degradation and reproduction problem due to an IWTP and MWTP effluents.     

水体pH和曝气方式对藻类生长的影响

利用水族箱微宇宙研究了藻类在不同pH和曝气条件下的生长和种类变化.使用天然湖水,一组试验每天调节pH,使其分别保持在8.0、8.5、9.0和9.5;另一组试验是设定不同的曝气方式,分别为不曝气、完全曝气、昼间曝气和夜间曝气,定期测定水体叶绿素a和藻类组成.pH试验结果显示,在pH 8.0～9.5范围内,pH 8.5下藻类生长状况最好,pH 9.5下生长最差,人为改变pH使其远离8.5能够抑制藻类生长.曝气试验结果显示,曝气不能抑制水体中藻类的生长,昼间曝气甚至还有明显的促进作用.     

Survival, growth and reproduction of Daphnia carinata (Crustacea: Cladocera) exposed to chronic cadmium stress at different food (Chlorella) levels

In nature, organisms have to respond to a diversity of factors acting simultaneously. The present investigation was conducted to study whether changes in food (Chlorella) levels could modify the chronic toxicity of cadmium on the various life-history parameters, such as survivorship, longevity, life expectancy, fecundity, age at first reproduction, R(0), T, r and growth rates of the cladoceran Daphnia carinata. The study indicated that at low food levels (0.5 x 10(6) cells ml(-1) Chlorella), cadmium concentrations in the range of 27-162 microg litre(-1) reduced these life-history parameters by 50% (EC(50)). At medium food levels (1.5 x 10(6) ml(-1) Chlorella) the EC(50) of cadmium was in the range of 51-127 microg litre(-1). At high food levels (4.5 x 10(6) cells ml(-1) Chlorella), the toxic effect of cadmium was greatly reduced. The decreases in survival, growth and reproduction of D. carinata at high cadmium-low food levels affected the fitness parameter 'r'. The study emphasises the need to include reproductive parameters other than mere survival in toxicity bioassays. The study also stresses the need to incorporate in laboratory tests other relevant factors that might modify pollutant toxicity.     

Effects of different food (Chlorella) concentrations on the chronic toxicity of cadmium to survivorship, growth and reproduction of Echinisca triserialis (Crustacea: Cladocera)

Extraneous factors have been shown to greatly modify pollutant stress. The present study was conducted with the objective of determining whether differences in food levels could modify chronic toxicity of cadmium to the various life-history parameters of the cladoceran Echinisca triserialis. Laboratory experiments were conducted on a sublethal range of cadmium (0, 2.5, 5.0, 10 and 20 microg litre(-1)) on life-history parameters such as survivorship, longevity, life expectancy, age at first reproduction, total fecundity, neonate size, net reproductive rate (R0), generation time (T), intrinsic rate of natural increase (r) and growth of Echinisca triserialis in relation to different food (Chlorella) levels of 0.5 (low), 1.5 (medium) and 4.5 (high) x 10(6) cells ml(-1). Cadmium levels of 10 microg litre(-1) and above, and low food levels, had a profound effect in decreasing the magnitude of all parameters studied. EC50 levels computed for life-history parameters, such as longevity, life expectancy at birth, total fecundity, R0 and T, were in the range of 2 to 21 microg litre(-1) cadmium, and this is indicative of extreme adverse effects on the population dynamics of E. triserialis when exposed to low food levels. At high food levels the EC50 was not achieved for cadmium in the toxicant range studied. The significance of these results is discussed in relation to field conditions.     

Short-term responses of Oryzias latipes (Pisces: Adrianichthyidae) and Macrobrachium nipponense (Crustacea: Palaemonidae) to municipal and pharmaceutical waste water in Beijing, China: survival, behaviour, biochemical biomarkers

Whole effluent toxicity was assessed for the fish Oryzias latipes and the prawn Macrobrachium nipponense for 18 h in a dilution series (0-66%) of the inflow and effluent of a municipal waste water treatment plant as well as waste water from a teramycin producing pharmaceutical industry, before, during and after a pilot laboratory purification process. The waste waters caused acute toxicity as measured by inhibition of light emission in the luminiscent bacterium Vibrio qingaiensis sp. nov. (Q67). EROD and aryl hydrocarbon hydroxylase activity in in vitro carp liver-cells showed a dose-dependent toxic response to the municipal waste water. Behavioural responses and time-to-death of fish and prawn, recorded online with the "Multispecies Freshwater Biomonitor" proved to be concentration- and time-dependent sensitive toxicity indicators in both types of waste water. Behaviour changed stepwise from normal activity to (increased or decreased) activity to more time spent on ventilation and finally to increased morbidity at higher concentration and time of exposure. The municipal waste water treatment plant managed to reduce toxicity to bacteria (Q67), prawn and fish. The pharmaceutical waste water treatment process still has to be improved, in order to reduce toxicity for fish and prawn.     

Effects of Cadmium and Zinc on the feeding behaviour of two freshwater crustaceans: Atyaephyra desmarestii (Decapoda) and Echinogammarus meridionalis (Amphipoda)

Lethal and sublethal effects of Cadmium and Zinc on two freshwater crustaceans were investigated. The 96-h LC50 for Cadmium and Zinc were 42.20 microg l(-1) and 5.43 mg l(-1) for Atyaephyra desmarestii, and 36.17 microg l(-1) and 4.61 mg l(-1) for Echinogammarus meridionalis, respectively. An increase in metal concentrations at sublethal levels resulted in significant reductions of the feeding rate of both species. The LOECs in the chronic feeding assays for Zinc were 1.29 mg l(-1) for A. desmarestii and 0.4 mg l(-1) for E. meridionalis. For Cadmium the LOEC was 6.53 microgl(-1) for both species. The results suggest the potential use of these two crustacean species as test organisms in ecotoxicological tests. Feeding assays appear to be a rapid, cheap and effective tool to be used in biomonitoring studies of Portuguese freshwaters.     

Production of male neonates in Daphnia magna (Cladocera, Crustacea) exposed to juvenile hormones and their analogs

We exposed the water flea Daphnia magna (Cladocera, Crustacea) to either juvenile hormone I (JH I), juvenile hormone II (JH II), or the juvenile hormone-mimicking insecticides kinoprene, hydroprene, epofenonane, or fenoxycarb. By 21-day reproduction tests, we investigated the effects on the number of neonates born per female and the offspring sex ratio. All six chemicals induced D. magna to produce male neonates; the male sex ratio of the offspring increased as the chemical concentration increased. EC50 values for production of male neonates were estimated as 400 (JH I), 410 (JH II), 190 (kinoprene), 2.9 (hydroprene), 64 (epofenonane), and 0.92 (fenoxycarb) microg/l. The number of neonates produced was reduced with all chemicals at the concentrations investigated. At the EC50 for male production, five of the six chemicals reduced the reproductive rate to less than 50%; the exception was epofenonane, which caused only a slight reduction in reproductive rate. These results were similar to those obtained for five juvenoids studied previously, one of which was studied here again. There are now 10 chemical substances--all juvenile hormones or their analogs-that are known to induce D. magna to produce male neonates. This suggests that juvenile hormone is involved in initiating male production followed by sexual reproduction in D. magna, and probably in most cladocerans that exhibit cyclic parthenogenesis.     

Responses of the mussel Anodontites trapesialis (Unionidae) to environmental stressors: effect of pH, temperature and metals on filtration rate

To evaluate the suitability of the tropical freshwater mussel Anodontites trapesialis for ecotoxicity assays, we tested the effects of temperature, pH, and Cd, Cu and Zn on its filtration rate. This is a relevant, sensible sublethal endpoint given the ecological role that this mussel plays in Amazonian environments. Filtration rate was calculated from the clearance of algae, fed to mussels at different temperature, pH and metal concentrations. Filtration rate was highest at 20 degrees C and pH 8, and decreased at low temperatures and pH. For all metals clear dose-response relationships were found. Cd exerted the most toxic effects (EC(50filtration) 64microg/L) followed by Cu (605microg/L) and Zn (4064microg/L). Metal mixtures representing present pollution levels clearly affected filtration rates. The results suggest that A. trapesialis is a suitable ecotoxicological test organism for the Amazonian region.