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Short Term Toxicity of Iron (Fe) and Lead (Pb) to the Mayfly Leptophlebiu Marginata (L.) (Insecta) in Relation to Freshwater Acidification

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Short Term Toxicity of Iron (Fe) and Lead (Pb) to the Mayfly Leptophlebiu Marginata (L.) (Insecta) in Relation to Freshwater Acidification Hydrobiologiu 284: 157-168, 1994. 0 1994 Kluwer Acndemic Pub1isher.r. Printed it, Belgium 157 Short term toxicity of iron (Fe) and lead (Pb) to the mayfly Leptophlebiu marginata (L.) (Insecta) in relation to freshwater acidification A. Gerhardt Lund University, Dept qf Ecology, Ecotoxicology, Helgonaviigen 5, S-22362 Lund, Sweden Received 18 March 1993; in revised form 22 July 1993; accepted 14 September 1993 Abstract The mayfly Leptophlebia marginata was exposed to different concentrations of Fe” or Pb2’ at pH 4.5 and pH 7.0. The effects of the metals on escape behavior and survival of the mayflies were investigated during an exposure of 120 hours. (1) Whole-body metal loads (Fe; Pb) of the mayflies increased in a dose-dependent way at both pH levels. A significant effect of pH on metal concentration in the mayflies was only found for Pb (p<O.OOl). (2) In terms of mortality, both metals were more toxic at pH 4.5 than at pH 7. The 96 h-LCS, val- ues for Fe were 106.3 mg Fe 1~ ’ at pH 7 and 89.5 mg Fe l- ’ at pH 4.5. Those for Pb were > 5 mg Pb11’atpH7and1.09mgPb111atpH4.5. (3) The mayflies lost their escape behavior, when exposed to the metals, the effects being more pro- nounced at low than at circumneutral pH for both metals (p < 0.05). The 96 h-EC,, values for Fe were 70.0 mg Fe 1~ ’ at pH 7 and 63.9 mg Fe 1~ ’ at pH 4.5. Introduction (Wren & Stephenson, 1991; Gerhardt, 1993). Some of the reasons for the contradictory results Lead (Pb) is a widely distributed contaminant. of the effects of pH on Pb toxicity may be differ- Lead concentrations in freshwater can reach up ences in water chemistry parameters and in the to 9 pg l- ’ (Jorgensen et al., 1991). Acidification tolerance of the various test species. of surface waters down to pH 5 causes an in- Iron (Fe) is an essential trace metal for all or- crease of aqueous Pb if the content of organic ganisms because it is a mediator in oxygen and ligands in the water is low (Nelson & Campbell, energy transport (Huebers, 1991). The concen- 1991). The presence of organic matter or amor- tration of iron in freshwater varies from 0.01 to phous Fe-hydroxides, however, reduces the avail- 1.4 mg Fe 1- ’ (Jorgensen et al., 1991). Only a few ability of Pb (Tessier et al., 1984). Investigations studies of Fe-toxicity have been performed, of the role of Pb in aquatic ecosystems have con- mostly without considering pH and Fe-speciation centrated on its bioaccumulation and biomagni- (Jorgensen et al., 1991; Gerhardt, 1993). There is, fication in the aquatic food web. Toxicity studies however, an indication that Fe may be more toxic have mostly been performed with fish, crusta- at low pH to Asellus aquaticus (Maltby et al., 1987) ceans, molluscs or insects as test organisms, how- and to Leptophlebia marginata (Gerhardt, 1992a). ever, the effect of pH on Pb toxicity is not uniform Mayflies are important links in the aquatic food 158 web. However, they are under-represented in metal concentrations (Pb2’: 0, 0.1, 0.5, 1.0 and studies on metal toxicity (Jorgensen et al., 1991; 5.0mgl-‘;Fe2’: 0, 10,50, 100,250 and 500 mg Gerhardt, 1993). Leptophlebia marginata and the 1-l) at both pH conditions. As the background closely related species Leptophlebia vespertina levels for Pb and Fe are IO.0 1 mg Pb l- ’ and occur at pH levels down to 4.0 and may be in- Ilmg Fe I-’ respectively, the concentrations creasing in abundance in acidified streams in used in the experiments were up to 500 times Sweden (0kland & 0kland, 1986). Since metal higher than in natural surface waters. The stream solubility and bioavailability increase at low pH, water in the aquaria was renewed daily and the these mayflies may be exposed to increased metal metal concentrations (Fe,,,, Fe* + , Pb,,,) were stress. As L. marginata is a particle feeder, it will determined before a fresh addition of the metal be exposed to metals with a tendency to bind to solutions (PbCl,; FeSO, x 7H,O) occurred. Fetot organic matter, such as Pb and Fe. and Pb,,, were determined with flame AAS, Mortality is one of the most frequently used whereas Fe* + was measured spectrophotometri- endpoint in toxicity studies. However, physiologi- tally at 522 nm using 2,2’ Bipyridine. The may- cal and behavioral responses to a toxicant are flies were not fed during the 120 h exposure pe- more sensitive (Warner, 1967) and in terms of riod. response time, they are among the first reactions Survival of the mayflies and their reaction to a against toxicant stress at sublethal doses (Beit- mechanical stimulus with a forceps (escape reac- inger, 1990). tion) were recorded daily. A positive escape re- To compare the lethal (mortality) and sublethal action was noted if the mayflies moved a distance (behavior) effects of Fe and Pb the mayfly Lep- of more than one body length away from the tophlebia marginata was exposed to the metals for stimulus, irrespective of the kind of movement 120 hours under circum-neutral and acidic con- (swimming, creeping). ditions. At the end of the experiments, alive animals from all treatment groups were prepared for metal analysis (Fetot, Pb,,,) with flame AAS. After rins- Material and methods ing with destilled water, the mayflies were dried to constant weight (48 h, 80 “C) and then digested Mayfly nymphs were collected from a small, soft- in HNO, suprapur at 80 “C (Gerhardt, 1990, water stream, rich in humic matter (Table 1). The 1992b). nymphs were kept in aquaria containing unfil- tered streamwater and stones serving as shelter. Statistical analysis For a period of two days, the animals were ac- climated to the system (10 “C+_ l”, 12 h/l2 h All data were analyzed by non-parametric statis- light/dark cycle) and to the two pH conditions of tical methods because of the lacking knowledge a) circum-neutral and b) pH 4.5 by stepwise ad- about the underlaying frequency distribution due justments twice a day with 0.1 M H,SO, and 0.1 to poor replication (r = 2). According to Manly M NaOH. (1991) an estimation of the distribution is only Two groups with 20 mayflies each were subse- useful at r2 10. Differences in the pH values be- quently exposed to one of the following nominal tween the various metal-concentration levels were Table 1. Chemical characterization of the water used for the experiments. ca’+ PH Cond. DOC NOi POi- *l,o, FPOM mgl-’ PScrn-’ mgC1 - ’ mgl-’ mgl- ’ mgl- ’ mgl- ’ 6.5 1.2 7.0 21.6 0.1 0.015 0.034 12.5 159 Table 2. pH and metal concentrations during the experiments. Group pH hot (md ‘1 Fe” (“/,) mean sd mean sd Fe control 4.64 0.17 1.3 0.2 n.d. lOmgl- 4.65 0.08 9.6 0.3 98 50 mgl~ ’ 4.56 0.10 47.0 2.8 97 100 mgl~ ’ 4.53 0.11 97.0 2.4 96 250 mgl - ’ 4.42 0.21 241.3 8.5 95 500 mgl - ’ 4.46 0.18 450 30 95 Fig. I. Uptake of Fe (mg g- ’ DW) by Leptophlebiu murginutu in relation to pH levels in the water. control 6.14 0.45 1.3 0.2 n.d. lOmgl-’ 6.51 0.32 9.4 0.3 93 50 mgl- ’ 6.42 0.34 45.8 2.0 94 cause it seems to be a more accurate method for 100 mgl- ’ 6.31 0.40 95.0 2.2 91 250 mgl- ’ 5.95 0.29 230.0 5.2 90 scattered data (Weber, 1986). 500 mgl - ’ 6.06 0.48 423.0 18 92 Results Pb control 4.60 0.2 0.002 0.0003 0.1 mgl-’ 4.61 0.15 0.11 0.02 Chemical parameters 0.5 mgl~ ’ 4.90 0.21 0.49 0.10 1.0 mgl- ’ 4.7 0.17 0.80 0.11 Between the daily renewals of the water and metal 5.0 mgl- ’ 4.6 0.19 4.10 0.35 solutions in the aquaria, the Fetot concentrations control 6.1 0.20 0.002 0.0003 were reduced by up to 10% at pH 4.5 and 20% 0.1 mgl~ ’ 6.5 0.454 0.09 0.02 at circumneutral pH and 2 90 y0 of the Fetot was 0.5 mgl - ’ 6.6 0.51 0.42 0.13 present as Fe2 + at both pH levels (Table 2). The 1.0 mgl- ’ 6.7 0.49 0.70 0.20 Pb concentrations decreased by 20% within 24 5.0 mgl- ’ 6.6 0.43 3.91 0.30 hours at both pH levels. At circumneutral pH, The values are means of 2 experimental units, with 10 mea- exposure to 500 mg Fe l- * caused a decrease in surements in each. pH down to pH 6.06, whereas in the correspond- ing acidified Fe-treatment, the mean pH values analysed with the Kruskal Wallis multiple com- were only slightly lower than in the controls parison test. The data for metal concentrations in (Table 2). However, neither in the Fe-experiments the mayfly nymphs from different metal treat- nor in the Pb experiments, the differences in pH ments were compared with Likelihood ratio tests. between the various metal exposures were signif- The Friedman test served as a non-parametric icant (p > 0.05, Kruskal Wallis).
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