Do Larvae and Ovipositing Chironomus riparius (Diptera: Chironomidae) Females Avoid Copper-Contaminated Environments?

Studies on the avoidance behavior of aquatic organisms to contaminants have confirmed that such behavior can be relevant in field situations. However, almost all toxicity tests involve the forced exposure of organisms to toxicants. In particular, despite the importance of Chironomus riparius Meigen larvae in sediment toxicity testing, only a few studies on avoidance behavior have been performed. This study investigated the ability of different life stages of C. riparius, including ovipositing females, first-, second-, and fourth-instar larvae, to avoid copper-contaminated environments. Ovipositing females were given a choice between a control and copper solution (1.3 mg Cu l^{? 1}). First-instar larvae were provided with a choice between a control and a copper (2.0 mg Cu l^{? 1})-spiked sediment. Both second- and fourth-instars were exposed to a copper gradient (0.38–3.4 mg Cu l^{? 1}) in a flow-through system. None of the life stages avoided copper, even though the highest concentrations caused lethal effects on midges. The avoidance behavior of C. riparius is not a sensitive endpoint to assess copper sublethal toxicity.     

Fluoranthene as a model toxicant in sediment studies with Chironomus riparius

The bioavailability of fluoranthene (as a model toxicant) in freshwater sediment tests with Chironomus riparius was investigated in relation to the partitioning of the chemical between water and sediment. Effects on survival and successful development of newly hatched larvae through to adult emergence were measured over a range of sediment concentrations. Radiolabelled fluoranthene was employed to enable sediment, aqueous and tissue concentrations to be determined. A significant effect on emergence was obtained at a measured sediment concentration of 80 mg kg^-1 (dry weight). The measured concentration in the overlying water was 88 g l^-1. Although effects at 170 mg kg^-1 (dry weight) were more severe, the measured concentration in the overlying water was similar (89 g l^-1), probably reflecting the approach towards the solubility limit. Bioaccumulation of fluoranthene, determined by analysis of emerging adults, was approximately proportional to the sediment concentration. Although the aqueous phase concentration appeared to be the principal factor controlling bioavailability, the increasing toxicity, when water concentrations had reached a plateau, suggested that other mechanisms may also be involved. Different sediment spiking procedures resulted in broadly similar partitioning of the chemical between the sediment and the overlying water. There were only minor differences in toxicity between the procedures, two of which allowed the carrier solvent used for spiking to be eliminated before addition of the test organisms. It was concluded that, for most purposes, all four procedures were equally acceptable, and by retaining a flexible approach, even for standardised regulatory tests, spiking procedures may be tailored to the physico-chemical properties of the test material.     

Nitrification in Freshwater Sediments as Influenced by Insect Larvae: Quantification by Microsensors and Fluorescence in Situ Hybridization

Sediment-reworking macrofauna can stimulate nitrification by increasing the O₂ penetration into sediments or it can reduce nitrification by grazing on nitrifying bacteria. We investigated the influence of Chironomus riparius larvae (Insecta: Diptera) on the in situ activity, abundance, and distribution of NH₄⁺-oxidizing (AOB) and NO₂^–-oxidizing bacteria (NOB) in two freshwater sediments with microsensors and fluorescence in situ hybridization. In organic-poor sediment, nitrification activity was reduced by the presence of C. riparius larvae, whereas no such effect was detected in organic-rich sediment. We explain this difference with the variable larval burrowing and grazing behavior in the two sediment types: In organic-poor sediment larval activities were intense and evenly distributed across the whole sediment surface, whereas in organic-rich sediment larval activities were locally restricted to the microenvironment of animal burrows. Surprisingly, the animals did not cause any significant change of the abundance of AOB and NOB. This implies that the observed reduction of nitrification activity was not density-regulated, but rather was due to the lowered metabolic activity of the nitrifiers. Partial digestion and redeposition of particle-associated bacteria by C. riparius larvae are believed to have caused this loss of metabolic activity.This revised version was published online in November 2004 with corrections to Volume 48.     

Biological effects of contaminated bottom sediments of water bodies in Central and South Vietnam on aquatic organisms

The levels of metal content and toxicity have been determined in bottom sediments of water bodies in Central and South Vietnam. It is found that the concentration of metals (As, Cr, Cu, Hg, Ni, Pb, and Zn) exceed the value of the abundance of elements in the Earth’s crust, which may be associated with geochemical features of the province. Acute toxicity of water-extracted sediment elutriate for Ceriodaphnia and whole (native) sediments for chironomid larvae is not recorded in any investigated water bodies. An increase in the relative abundance of Chironomus riparius larvae with deformations of mouthpart structures compared to the control was observed for most of the water bodies. This index reaches the value in the Khe River (Cân Th? city). Antenna anomalies prevailed in pathomorphological disorders.     

Bioavailability of zinc in the sediment to the estuarine amphipod <Emphasis Type="Italic">Grandidierella</Emphasis><Emphasis Type="Italic">japonica</Emphasis>

The utility of interstitial water concentrations of metals and simultaneously extracted metals/acid-volatile sulfide differences (SEM–AVS) in two seasons were investigated to explain the biological availability of zinc in sediments to benthic organisms exposed in the laboratory. The amphipod Grandidierella japonica was exposed, in 10-day acute toxicity tests, to clean sediment spiked with zinc to obtain nominal treatments ranging from 0.25 to 74.4 mol g^–1 dry weight with respect to the molar difference between SEM_Zn and AVS. When the molar difference between SEM_Zn and AVS (i.e., SEM–AVS) was <0 mol g^–1, the concentration of zinc in the sediment interstitial water was low and few adverse effects were observed for any of the biological endpoints measured. Conversely, when SEM_Zn–AVS exceeded 0 mol g^–1, the concentration of zinc in the interstitial water and amphipod mortality increased. These data compare favorably with observations made in short-term exposures and thus support the use of AVS as a normalization phase for predicting toxicity in metal-contaminated sediments in different season.     

Effects of Cu,Pb and Zn on two Potamogeton species grown under field conditions

Two common macrophyte species, Potamogeton perfoliatus L. and Potamogeton pectinatus L. were grown for 12 weeks at shallow depths in sediments contaminated with 1250 or 2500 g Pb or Cu and/or Zn (gDW sediment)^-1. Control experiments were run at background levels of 4, 13, and 38 g Pb, Cu and Zn (gDW sediment)^-1, respectively. Effects of heavy metals on biomass production and metal uptake and distribution in plants are presented in relation to total amount and plant-available fraction of metals in the sediment.All three studied metals gave reduced biomass production, and the toxicity of the metals decreased in the order Zn>Cu>Pb. The root/shoot biomass ratio increased for P. pectinatus, but decreased for P. perfoliatus with metal treatment. The content of any single metal was higher in shoots than in roots of plants grown on sediments not contaminated with that specific metal, but addition of that metal increased the proportion in roots. The uptake by plants of any of the heavy metals increased with increased metal addition. The magnitude of the plant-available fraction of metals of untreated sediment was Zn>Cu>Pb, and increased in contaminated sediments. Addition of Cu decreased both the plant-available fraction and the total concentration of Zn in the sediment, while increased the uptake of Zn by the plants. The opposite was found for Cu when Zn was added. P. pectinatus accumulated about twice as much Cu as P. perfoliatus. On the other hand, the concentration of Pb was higher in P. perfoliatus than in P. pectinatus, and was negligible in P. pectinatus when cultivated in untreated sediments.     

Enhanced exoenzyme activities in sediments in the presence of deposit-feeding Chironomus riparius larvae

1. The combined effects of deposit‐feeding, bioturbation and bioirrigation by benthic macrofauna on the enzymatic hydrolysis of organic matter were studied in microcosms. Chironomus riparius larvae (Insecta, Diptera) served as model macrofauna and stinging nettle leaves (Urtica dioica) were used as a detrital food source. 2. In the upper 10 mm of the sediment (the habitat of C. riparius larvae), the activities of several exoenzymes, the contents of several fractions of particulate organic matter (POM), and the concentrations of dissolved oxidants (O₂, NO) were measured on a small scale. Fluorescent particles (luminophores) were used to quantify the vertical redistribution of particles within the same layer. 3. In control sediment, the addition of detrital food enhanced exoenzyme activities in the 0–2 mm layer only. In the presence of C. riparius larvae, exoenzyme activities increased to 10 mm depth. Further, the content of POM in the 0–2 mm layer was lower in the presence than in the absence of larvae, suggesting ingestion and subduction of the added detritus. After prolonged incubation without further food addition, exoenzyme activities returned close to background values in both treatments, whereas the vertical distribution of POM remained unchanged. 4. The overall penetration depth of O₂ and NO into the sediment was greater in the presence than the absence of C. riparius, the differences being more pronounced after prolonged incubation. Locally high O₂ and NO concentrations due to bioirrigation by C. riparius were measured deep in the sediment. Net downward transport of particles was observed only in the presence of C. riparius larvae and only at the beginning of the incubation. 5. I conclude that deposit‐feeding and bioturbation by macrofauna can quickly remove freshly deposited POM from the sediment surface and transfer it to less oxygenated sites (i.e. animal guts and deep sediment layers). Bioirrigation also increases the availability of oxidants deep in the sediment. The oscillation of oxidant supply to POM particles by ingestion–egestion, burial and re‐burial, and the intermittent bioirrigation of subsurface sediment, is probably the cause of the increased rate of organic matter hydrolysis, the rate‐limiting step in mineralization.     

Oviposition habitat selection in response to risk of predation in temporary pools: mode of detection and consistency across experimental venue

Natural selection should favor females that avoid ovipositing where risk of predation is high for their progeny. Despite the large consequences of such oviposition behavior for individual fitness, population dynamics, and community structure, relatively few studies have tested for this behavior. Moreover, these studies have rarely assessed the mode of detection of predators, compared responses in prey species that vary in vulnerability to predators, or tested for the behavior in natural habitats. In an outdoor artificial pool experiment, we tested the oviposition responses of two dipteran species, Culiseta longiareolata (mosquito) and Chironomus riparius (midge), to the hemipteran predator, Notonecta maculata. Both dipteran species have similar life history characteristics, but Culiseta longiareolata larvae are highly vulnerable to predation by Notonecta, while Chironomus riparius larvae are not. As their vulnerabilities would suggest, Culiseta longiareolata, but not Chironomus riparius, strongly avoided ovipositing in pools containing Notonecta. An experiment in natural rock pools assessing oviposition by Culiseta longiareolata in response to Notonecta maculata yielded an oviposition pattern highly consistent with that of the artificial pool experiment. We also demonstrated that the cue for oviposition avoidance by Culiseta longiareolata was a predator-released chemical: Notonecta water (without Notonecta replenishment) repelled oviposition for 8 days. Oviposition avoidance and mode of detection of the predator have important implications for how to assess the true impact of predators and for the use of commercially produced kairomones for mosquito control.     

Chronic toxicity of cadmium toChironomus riparius Meigen — effects upon larval development and adult emergence

Chronic cadmium toxicity tests were carried out withChironomus riparius Meigen using a static-with-replacement exposure procedure. Significant reductions in larval development, survival and production were detected at 0.15 mg Cd l^–1 a value approximately 5000 times lower than the 48 h LC50 for fourth instar larvae. The first instar appears to be the most critical stage in terms of sensitivity, with the number of adults eventually emerging reflecting first instar mortality. Male emergence was slightly delayed at 0.15 mg Cd l^–1. The results of this study illustrate the importance of chronic toxicity tests in identifying sensitive life cycle stages and explaining pollutant-induced stress.     

Physiological energetics of a midge,Chironomus riparius Meigen (Insecta,Diptera): normoxic heat output over the whole life cycle and response of larva to hypoxia and anoxia

The rate of metabolism of laboratory reared Chironomus riparius was monitored by direct calorimetry over the entire life cycle from egg to adult stage. The metabolic response of the fourth instar larva to decreasing oxygen concentrations and anoxia was also measured. Normoxic measurements were carried out at 20°C and the hypoxic-anoxic experiments at 10°C. In larvae with body sizes ranging from 0.0028 to 0.645 mg ash-free dry mass (afdm), the rate of heat dissipation was related to body mass by a power function, with a mass exponent of 0.71±0.02 corresponding to an exponent of -0.29 for the relationship between mass-specific metabolic rate and body mass. However, the allometric equations applicable to larvae would not predict the metabolic rates of eggs, pupae and adults. Single egg batches used in the experiments consisted of 354±90 eggs, the individual egg with a mass of 0.99±0.01 g (mean±SD). The mass-specific rate of heat dissipation of the egg (13.7±1.8 W mg^-1 afdm) was considerably lower than that of the first and second instar larvae (44–53 W mg^-1) but equal to that of fourth instar larvae (13.1±3.9 W mg^-1). Heat dissipation by a pupa shortly before adult emergence was high (14.8±1.8 W mg^-1), probably due to high metabolism during metamorphosis. Emergence of the adult in the calorimeter was indicated by a short but intense burst of heat. The newly emerged imago had a ca. 20–35% higher metabolic rate than the pupa. In response to reduced O₂ partial pressure the fourth instar larva of C. riparius displayed metabolic regulation. In continuously declining oxygen partial pressure, the fourth instar larva maintained its aerobic energy metabolism (4.2 W mg^-1) with only a small decrease down to 0.8 kPa, corresponding to an oxygen concentration of 0.42 mg O₂l^-1 H₂O. Below this critical oxygen concentration (P_c), the rate of heat dissipation decreased rapidly down to the anoxic level which was only 14–17% of the normoxic level. The high relative reduction of metabolic rate under anoxia gives a wrong impression of short-term tolerance of C. riparius to anoxia. The absolute energetic costs of C. riparius associated with anaerobic energy metabolism (0.64±0.11 W mg^-1) are almost 6 times higher than those of more anoxia tolerant invertebrates such as sphaeriid bivalves.     

Toxicological and teratogenic assessment of bottom sediments from the Rybinsk Reservoir

The toxicity of bottom sediments (BSs) from different parts of the Rybinsk Reservoir has been determined by biotesting. Neither acute nor chronic toxicity are detected for ceriodaphnia Ceriodaphnia affinis Lillijeborg in tests with water extracts of sediments from all the investigated stations. The relative number of Chironomus riparius Meigen larvae with deformed mouth structures in the specimens exposed to the BSs of the reservoir is higher than in the control. Abnormalities of the mentum and mandibules (heavily chitinized structures) were the most often observed pathomorphological disturbances. The results of this study suggest that BSs from the stations located near the Cherepovets Industrial Complex (CIC) may be classified as contaminated.     

Impact of a diazinon calamity in 1996 on the aquatic macroinvertebrates in the River Meuse, The Netherlands

A more or less continuous load of pesticides has been noted in the River Meuse in recent years. In April 1996, when high concentrations (up toca. 1 g 1^–1) of the insecticide diazinon were measured in the River Meuse at the Belgian-Dutch border, the maximum concentration for drinking water production was exceeded. This was alerted after activity changes of fish and daphnids in the biological monitoring systems (RIZA). These observations were compared with literature toxicity data of diazinon, in order to determine the ecological impact of this diazinon discharge on the aquatic macroinvertebrates. LC₅₀ values of several aquatic macroinvertebrate species were exceeded. In addition, a high mortality was observed in bioassays with the midgeChironomus riparius and the caddisflyHydropsyche angustipennis. It is inferred that the species composition of the macrofauna community in the River Meuse is likely to be reduced by such an accident. Recolonization of the River Meuse by sensitive macrofauna species may therefore be prevented by incidental pesticide discharges.     

The acute toxicity of cadmium to different larval stages of Chironomus riparius (Diptera: Chironomidae) and its ecological significance for pollution regulation

Summary Acute toxicity tests using a static-with-replacement testing procedure were carried out with the four larval instars of the freshwater detritivore Chironomus riparius (Meigen). Median lethal concentrations (10, 24, 48 and 96 h LC50's) indicated great differences in sensitivity to cadmium between instars. Larvae became more tolerant with increasing age, the most resistant stage (fourth instar) having a 24 h LC50 of 2,400 mg Cd l^-1, approximately 950 times greater than the corresponding value of 2.1 mg Cd l^-1 recorded for the most sensitive (first instar) stage. These observations are discussed in relation to the establishment of water quality standards.     

The relationship between sediment mobilisation and the entry of Baetis mayflies into the water column in a laboratory flume

Mass bedload movement is thought to play a key role in initiating stream invertebrate drift during extreme flood events. However, little is known of the importance of the shear of invertebrates from stone surfaces relative to their entrainment along with bed material at different discharges. In particular, it is unclear whether so-called catastrophic drift only occurs once mass bedload movement, and hence entrainment of invertebrates, occurs. We investigated the relationship between the mobilisation and transport of bed sediments and the entry of Baetis mayflies into the water column in a laboratory flume. Experiments quantified the percentage of Baetis drifting at a range of discharges that mobilised between 0 and 95% of the flume-bed sediments. Control experiments quantified drift losses from sediment fixed to the bed of the flume, such that sediments were immobile even at the highest discharges. Drift losses increased with increasing discharge and velocity in the flume. Sediment mobility contributed significantly to drift (ANCOVA, p < 0.001), with consistently greater drift losses in mobile sediment experiments than in those with fixed sediment. The discharge which resulted in a loss of 100% of Baetis from the mobile sediment bed (discharge 30 l s^–1) resulted in a loss of approximately 50% of individuals from the fixed bed. Results indicate that once bed sediments are mobilised, entry of Baetis into the drift is greater than expected from the shear of animals from stone surfaces alone. Thus, entrainment of animals along with sediment contributes significantly to drift at high flows. This implies that differences in bed stability between sites or streams, or temporal changes in sediment characteristics within a site, could influence patterns of drift.     

Transport of carbon dioxide and ammonium in bioturbated (Nereis diversicolor) coastal, marine sediments

Two identical experiments with sieved and homogenized sandy and muddy sediment were conducted to determine transport enhancement of porewater solutes (TCO₂ and NH₄ ⁺) by the presence of the polychaeteNereis diversicolor (1000–1500 m^–2). Flux measurements showed thatN. diversicolor enhanced the release of CO₂ and NH₄ ⁺ 1.5–5 times. Accordingly, porewater concentrations of these compounds were reduced considerably in the bioturbated zone of both types of sediments. Two different diagenetic models, effective (eddy) diffusion and nonlocal exchange, were used to describe solute profiles in the bioturbated sediments. In permeable sandy sediments advective porewater movements may occur more readily than in more cohesive muddy sediments. The effective diffusion model (with D_e=1.6–2.0 cm² d^–1) provided an excellent fit to the measured concentrations of both solutes below the bioturbated zone in permeable sandy sediment, whereas this model overestimated the concentration in the bioturbated zone. However, in the less permeable muddy sediment the effective diffusion model overestimated the NH₄ ⁺ profile considerably at all depths. The nonlocal exchange model (with=0.17–0.29 d^–1), on the other hand, provided an excellent fit in the less permeable muddy sediment, suggesting that solute profiles here were controlled by molecular diffusion, even in the presence of burrow irrigation. For the permeable sediment, the nonlocal exchange model (with=0.14 d^–1) underestimated the measured NH₄ ⁺ profile. Accordingly, linear slopes from plots of porewater TCO₂ as a function of porewater NH₄ ⁺ revealed that eddy diffusion (or advective porewater movements) was important in the bioturbated zone of this sediment type. However, combined with the generally more realistic shape of profiles derived by the nonlocal exchange, these evidences suggest that both eddy and molecular diffusion must operate in the bioturbated zone of permeable sediments.     

Continuously measured changes in redox potential influenced by oxygen penetrating from burrows of Callianassa subterranea

A continuously recording multiple redox sensor was used to measure oxygenation effects in otherwise anoxic sediments. The sensor reacted within three minutes (increase > 50 mV) to oxygen penetrating into the sediment. Different patterns of oxidation/reduction were observed along burrows of sediment-dwelling infauna. Examples of an increase of more than 600 mV lasting 50 hours and short, 1 to 4 hours long, oxidation pulses of 280 mV increase are given. Redox patterns similar to the ones observed along natural burrows were generated using artificial burrows. Oxygen penetrated up to 1 mm by diffusion from burrow walls into the adjacent sediment, whereas elevated Eh values were measured within 3 mm of the burrow. The oxygenation effect of a burrowing shrimp, Callianassa subterranea, was studied in a 3-month experiment with ten sensors and five animals. For one individual we recorded 108 events with Eh increasing by more than 300 mV. It was estimated that below every square meter of sediment surface 0.7 m². burrow surface are subjected to oxygenation at least once per day by the population of Callianassa in the southern North Sea.     

The impact of sediment reworking by opportunistic chironomids on specialised mayflies

1. Bioturbation, by definition, changes the structure and properties of sediments, thereby altering the environment of the bioturbator and other benthic species. In addition to the indirect effects of sediment reworking (e.g. changes in water quality), bioturbating species may also directly interfere with other species via competition. This study aims, therefore, to examine both the direct and indirect effects of sediment reworking by an opportunistic detritivore on survival and growth of a specialised mayfly species. 2. Bioturbation was imposed by adding different densities of the midge Chironomus riparius to clean and polluted sediments. Changes in water quality and sediment properties, and survival and growth of the mayfly Ephoron virgo were assessed. 3. Chironomid density had a strong negative effect on the concentrations of metals, nutrients and particles in the overlying water, but increased the penetration of oxygen into the sediment. Survival and growth of E. virgo were strongly reduced in the presence of chironomids. In the polluted sediment, the activity of chironomids enhanced the negative effects of pollution on E. virgo. In the clean sediment, inhibition of the mayfly was even more pronounced. 4. This suggests that direct disturbance by C. riparius was more important than indirect changes in water quality, and over‐ruled the potential positive effects of improved oxygen penetration. The results indicated that the distribution of small insects, such as E. virgo, can be limited by bioturbating benthic invertebrates.     

Gliding motility of Peloploca spp., and their distribution in the sediment and water column of a eutrophic lake

Observations indicating gliding motility in the gas-vacuolate, filamentous organism Peloploca were made using microcapillary tubes. Tubes containing semi-solid agar, incubated in sediment cores gave good enrichments of Peloploca spp. The organisms, which had the form of helical bundles of filaments, were seen to corkscrew through the agar at up to 2–3 m s^-1.The vertical distribution of Peloploca spp. in the sediment and water column of a eutrophic lake was examined periodically during summer stratification. The organisms were confined to anoxic conditions in the sediment prior to stratification. With increasing anoxia in the hypolimnion, the population shifted upwards in the sediment, and towards the end of stratification, in the most reducing conditions, appeared in the lower hypolimnion. Anaerobically incubated sediment cores also showed the movement of the Peloploca population from sediment into the overlying water.It is suggested that the gliding motility and helical morphology of Peloploca bundles enable them to migrate through sediment in response to oxygen and Eh gradients, in addition to their use of gas vacuoles to adjust their position in the water column. The taxonomic implications of gliding motility in Peloploca spp. are discussed.     

Influence of nitrogen species (NH4 + and NO3 −) on the dynamics of P in water–sediment–Salvinia herzogii systems

Water – Salvinia herzogii – sediment systems were exposed to different phosphorus and nitrogen combinations in outdoor experiments. The aim was to estimate the amounts of P immobilized in macrophytes and sediments, as well as to elucidate whether or not the presence of N affects the retention of P. The following components were added: o-P, o-P + NH₄ ⁺, o-P + NO₃ ^– + NH₄ ⁺, o-P + NO₃ ^–. The concentration of nutrients was periodically determined throughout the experiment (28 days). The concentrations of P and N in plant tissues and sediments were determined at the beginning and the end of the experiment. Sequential extractions of P-fractions in sediment were performed using the EDTA method (Golterman, 1996). The removal efficiency of P in water was 95–99%. The removal of NH₄ ⁺ (97–98%) was more effective than that of NO₃ ^– (44–86%). The presence of nitrogen species increased the removal velocity of o-P from water, NH₄ ⁺ was the most effective species. Sediments not only had higher P removal rates than macrophytes but, in the control treatment without macrophytes, they reached the values obtained by macrophytes plus sediments in the other treatments. The adsorption of P takes place at the surface layer of the sediment (1 cm). Most of the P incorporated into the sediment during the experiment was sorbed by the fraction Fe(OOH)P. The addition of nutrients to water modified the leaves/lacinias weight ratio.     

Effect of artificial destratification on iron,manganese, and zinc in the water,sediments, and two species of benthic macroinvertebrates in an Oklahoma lake

Ham's Lake was destratified in the summers of 1977 and 1978 by pumping surface water to the bottom. However, an arm was prevented from mixing by a submerged dam of a former farm pond and remained stratified. The concentration of manganese in the water increased to over 5 000 µg 1^–1 in the hypolimnion of the anoxic arm in summer. Soluble manganese averaged 28% of total concentration. Total iron concentrations were relatively high in the water and sorbed iron values were extremely low in the sediments. Soluble iron averaged 12% of total concentration. Zinc values were low in the water and sediments. Chaoborus punctipennis and Chironomus riparius concentrated iron and zinc and discriminated against manganese. Concentration factors were unaffected by mixing.