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Contrasting Sensitivities to Toxicants of the Freshwater Amphipods Gammarus Pulex and G

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Contrasting Sensitivities to Toxicants of the Freshwater Amphipods Gammarus Pulex and G View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Wageningen University & Research Publications Ecotoxicology (2010) 19:133–140 DOI 10.1007/s10646-009-0398-y Contrasting sensitivities to toxicants of the freshwater amphipods Gammarus pulex and G. fossarum A´ lvaro Alonso Æ Hendrika J. De Lange Æ Edwin T. H. M. Peeters Accepted: 29 July 2009 / Published online: 21 August 2009 Ó Springer Science+Business Media, LLC 2009 Abstract Amphipods are an important component of tolerant to cadmium than adults. In the case of ivermectin, freshwater ecosystems. They are very often used in eco- only for G. fossarum EC50 values were different between toxicology, particularly the freshwater amphipod Gamma- life stages. These results suggest that the risk assessment of rus pulex. However, there is scarce information on the toxicants to freshwater amphipods should include bioas- sensitivity to toxicants of other species within the genus says with the most sensitive species and life stage. Gammarus. The present study aims to: (1) to compare sensitivities to ivermectin and cadmium between two spe- Keywords Cadmium Á Ivermectin Á Juveniles Á cies of freshwater amphipods (G. pulex and G. fossarum); Adults Á Toxicity (2) to compare sensitivities to these toxicants between juveniles and adults within each species; and (3) to assess whether the sensitivity to toxicants of these co-generic species is related with the wideness of their natural distri- Introduction bution area. Eight independent short-term bioassays (96 h) were conducted to assess sensitivity for ivermectin and Amphipods are an important component of freshwater cadmium for juvenile and adult life stages for each species. ecosystems since they play a key role in the detritus The LC50 (mortality) and EC50 (mortality plus immobil- breakdown process and constitute an important source of ity) were calculated to 48 and 96 h of continuous exposure. food for predators (Forrow and Maltby 2000; MacNeil Our results showed that G. pulex was less tolerant to et al. 2000). They can be found in high densities and may ivermectin than G. fossarum, the reverse being true for be very sensitive to a wide range of toxicants (Felten et al. cadmium. In general, juveniles of both species were less 2008). These reasons explain why amphipods are often used in ecotoxicology. The most widely employed amphipod is the common species Gammarus pulex (L.) (Crustacea, Amphipoda), principally because this species has a wide distribution area (Barnard and Barnard 1983). A´ . Alonso (&) Other Gammarus species, like G. fossarum (Koch), are ´ ´ Departamento de Ecologıa, Facultad de Biologıa, Universidad de comparatively less employed in ecotoxicology mainly due Alcala´, Alcala´ de Henares, 28871 Madrid, Spain e-mail: [email protected] to their narrow distribution range. Furthermore, the avail- able information on the sensitivities of these other A´ . Alonso Á E. T. H. M. Peeters Gammarus species to toxicants is scarce compared to Aquatic Ecology and Water Quality Management Group, G. pulex. Gammarus pulex and G. fossarum occur both in Wageningen University, P.O. Box 47, 6700 AA Wageningen, The Netherlands Dutch streams with G. pulex having a much wider geo- graphical distribution than G. fossarum (Peeters and H. J. De Lange Gardeniers 1998). Optimum ranges for several common Centre for Ecosystem Studies, Alterra, Wageningen University water quality parameters, such as current flow, stream and Research Centre, P.O. Box 47, 6700 AA Wageningen, The Netherlands dimensions, chloride, conductivity, nutrients and dissolved 123 134 A´ . Alonso et al. oxygen levels are much narrower for G. fossarum than for Materials and methods G. pulex (Peeters and Gardeniers 1998). Thus we hypoth- esize that G. fossarum is more sensitive to toxicants than Amphipod collection and laboratory acclimatization G. pulex. Since the selection of the most sensitive stage permits a Gammarus pulex was collected using two sieves (2 and more suitable risk assessment of toxicants (McCahon and 4 mm mesh size) from a relatively unpolluted reach of the Pascoe 1988), it is of important ecotoxicological concern Heelsum Stream (51° 580N, 5° 450E) near Wageningen whether sensitivity to toxicants depends on life stages. (The Netherlands) (De Lange et al. 2006a; b). Gammarus According to McKim (1977) early fish stages (larval and fossarum was collected using a dip-net from two relatively early juvenile stages) are usually the most sensitive to the unpolluted upper streams (Geul Partij 50° 480N, 5° 510E, toxicity of different compounds. However, studies on Cottesen 50° 450N, 5° 560E) near Maastricht (The Nether- freshwater macroinvertebrates have shown contradictory lands), where cadmium concentrations were less than results with earlier stages of the dipteran Chironomus 0.4 lg/l (data from the regional Waterboard Roer en riparius and the crustacean Asellus aquaticus, being more Overmaas, The Netherlands). Invertebrates were trans- sensitive to certain metals than adults (Williams et al. ferred to the laboratory using plastic containers (5 l), and 1986; Naylor et al. 1990), and no difference in sensitivity kept in aerated aquaria in a temperature and humidity for the mollusc Potamopyrgus antipodarum to ammonia controlled room (15 ± 1°C; 60% humidity). Animals were between contrasting sizes (Alonso and Camargo 2004a). fed with stream-conditioned poplar (Populus sp.) leaves Furthermore, adults of the crustacean A. aquaticus were and were progressively acclimatized to the test water more sensitive than juveniles to low dissolved oxygen and (Dutch Standard Water-DSW; Netherlands Normalisation ammonia (Maltby 1995). The literature also reports con- Institute 1980) during at least 1 week prior to the bioas- trasting results for G. pulex with higher sensitivities to says. This standarized water was used because it has sim- cadmium for juveniles, and higher sensitivity of adults to ilar physical–chemical properties as most natural Dutch ammonia and low dissolved oxygen (McCahon and Pascoe freshwaters (Netherlands Normalisation Institute 1980). 1988; Maltby 1995). The latter result can be partly For each species two life stages were selected on the basis explained by intraspecific differences in ventilation rate of body length: adults (pre-copulatory pairs were rejected) (Maltby 1995). Regarding G. fossarum, to our knowledge and juveniles (McCahon and Pascoe 1988; Pockl 1992). no data are available to the sensitivities between contrast- ing life stages so far. Therefore, for each toxicant and Bioassay design species, specific bioassays are necessary to know the rel- ative tolerance of each stage. In the last decades many Adults and juveniles of G. pulex and G. fossarum were studies on rearing and handling of amphipods have been used to test the effects of ivermectin and Cd in independent published (McCahon and Pascoe 1988; Morritt and Spicer bioassays that lasted for 4 days. Independent bioassays 1996), facilitating the use of different life stages for labo- were used to avoid possible cannibalism of adults over ratory bioassays. juveniles, or competition between different species. Toxic The aims of the present study are (1) to test experi- solutions (Cd and Ivermectin) and controls were renewed mentally the hypothesis that G. fossarum is more sensitive every 2 days. Amphipods were not fed during the to toxicants than G. pulex, and (2) to evaluate how sensi- bioassays. tivities to these compounds vary between adults and Design of bioassays is shown in Table 1. For iver- juveniles for both species. We used two toxicants (iver- mectin bioassays, each amphipod was individually placed mectin and cadmium) that differ in their mode of action. in a glass vessel with 40 ml of toxic solution, acetone Ivermectin is a synthetic antiparasitic compound supplied control (acetone concentration = 0.0055 ml acetone/l), or to domestic cattle on a regular basis. This compound control water (Dutch Standard Water-DSW, Netherlands amplifies the effect of glutamate on the chloride channels Normalisation Institute 1980). For each treatment a total of invertebrates, causing their opening (Rohrer and Arena of 30 individuals were used. In each treatment, each group 1995). Cadmium is a nonessential heavy metal which can of 10 individuals was considered as a replicate to calculate be accumulated on gills and hepatopancreas of crustaceans the percentage of individuals showing a response. There- causing damage on cells and disruption of enzymatic fore, three replicates (i.e. three groups of 10 individuals reactions (Felten et al. 2008). Several studies have shown per group) were used for each treatment to calculate LC that crustaceans are very sensitive to both compounds and EC values. All vessels were covered with a plastic foil (Garric et al. 2007; Felten et al. 2008). The high sensitivity to reduce water evaporation. As ivermectin is known and the contrasting mode of action were the criteria used to to be photodegradable (Garric et al. 2007), bioassays select these toxicants. were conducted in darkness, in a temperature and 123 Contrasting sensitivities to toxicants of the freshwater amphipods 135 Table 1 Experimental designs for the bioassays with Gammarus pulex and Gammarus fossarum Toxicant Species Stage Length (±SD) Nominal concentrations Solution Individuals Controls (mm) (n = 40) volume (ml) per vessel Ivermectin G. pulex Adults 11.01 ± 1.55 1.5, 2.5, 3.5, 4.5, 5.5 lg/l 40 1 Acetone (0.0055 ml/l) and DSW G. pulex Juveniles 4.70 ± 0.88 1.5, 2.5, 3.5, 4.5, 5.5 lg/l 40 1 Acetone (0.0055 ml/l) and DSW G. fossarum Adults 10.11 ± 0.94 1.5, 2.5, 3.5, 4.5, 5.5 lg/l 40 1 Acetone (0.0055 ml/l) and DSW G. fossarum Juveniles 5.06 ± 0.90 1.5, 2.5, 3.5, 4.5, 5.5 lg/l 40 1 Acetone (0.0055 ml/l) and DSW Cadmium G. pulex Adults 10.6 ± 1.2 0.3, 0.5, 0.7, 1.0, 2.0 mg/l 300 10 DSW G. pulex Juveniles 4.0 ± 0.7 0.1, 0.3, 0.5, 0.7, 1.0 mg/l 20 10 DSW G.
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