Life cycle responses of the midge Chironomus riparius to polycyclic aromatic compound exposure

During acute exposure, polycyclic aromatic compounds (PACs) act mainly by narcosis, but during chronic exposure the same compounds may exert sublethal life cycle effects. The aim of this study was therefore to evaluate the chronic effects of sediment spiked PACs on the emergence of the midge Chironomus riparius. Three isomer pairs were selected, and 28-day LC50 values and 50% emergence times (EMt50) were determined. Concentration-response relationships were observed for phenanthrene, acridine, phenanthridine and acridone. Anthracene and phenanthridone had no effect on total emergence, but did cause a delay in emergence. Calculated porewater LC50 values correlated well with logKow values, suggesting narcosis as mode of action. In contrast, effect concentrations for delay in emergence (EMt50) deviated from narcosis, suggesting a specific mode of action during chronic exposure. It is concluded that emergence is a powerful endpoint to detect life cycle effects of PACs on a key sediment inhabiting invertebrate.     

Combined effects of copper and food on the midge Chironomus riparius in whole-sediment bioassays

Effects observed in whole-sediment bioassays must be seen as the joint effect of all sediment characteristics. In whole-sediment bioassays, however, adverse effects on test organisms are usually attributed to the presence of contaminants and effects of food are often ignored. The aim of this study was to analyze the response of the midge Chironomus riparius to sediment spiked with different combinations of food and copper. The responses of C. riparius to these spiked sediments were assessed in 10-day whole-sediment bioassays. Decreases in survival, dry weight, and length of C. riparius were observed with increasing copper concentrations. However, an increase in the amount of food resulted in an increase of larval dry weight and length until copper concentrations reached a critical threshold of 200 mg/kg. In addition, an increase in the amount of food resulted in a decrease of accumulated copper in the larvae. The present study demonstrated that the combination of copper and food in the sediment determines the performance of C. riparius in whole-sediment bioassays. The dependency of C. riparius on high feeding levels, which mask toxic effects, questions its suitability as a test organism for whole-sediment bioassays. Because benthic communities in polluted ecosystems are often exposed to varying levels of both food and toxicants it is concluded that the trophic state of the ecosystem may alter the ecological risk of sediment-bound toxicants to opportunistic benthic invertebrates such as C. riparius.     

Comparative metabolism of phenanthridine by carp (Cyprinus carpio) and midge larvae (Chironomus riparius)

Abiotic transformation of azaarenes in the environment has been analysed extensively, but metabolism is less well described. To further elucidate preliminary observations of interspecific differences in azaarene metabolism by aquatic organisms, phenanthridine biotransformation by midge larvae and carp was studied. In both experiments, 6(5H)-phenanthridinone (phenanthridone) was found as an important metabolite. The fish were clearly capable of metabolising phenanthridine, but in the midge experiment the metabolite was principally formed by bacteria growing on the food and not by the midges. Phenanthridone itself was further degraded to non-observed compounds in both experiments, due to bacteria and midges acting together in the midge experiment, and by carp in the fish experiment. Internal concentrations of phenanthridine and phenanthridone were non-detectable in the midge larvae, but concentrations of both compounds in carp organs suggested a major role of bile and liver. Since phenanthridone did not account for all phenanthridine loss, it was suggested that, apart from phenanthridone degradation, other metabolic pathways may play a role. This study clearly demonstrates the importance of interspecies differences in metabolism, which should not be neglected in risk assessment.     

Induction of mouthpart deformities in Chironomus riparius larvae exposed to 4-n-nonylphenol

Chironomid mouthpart deformities have often been associated with sediment contamination and are, therefore, currently used to assess sediment quality. Deformities were only occasionally induced in laboratory bioassays. Mouthpart deformities results from a physiological disturbance during larval molting. In the past few years it has been shown that some chemicals can exert negative effects on both vertebrates and invertebrates at the level of endocrine regulation. As insect molting is hormonally regulated, we wanted to test the hypothesis that deformities are induced due to a hormonal disruption in the developmental process. The aim of the present study was to test whether the endocrine disrupter, 4-n-nonylphenol (4NP), induces mouthpart deformities in chironomids. A laboratory bioassay was performed exposing Chironomus riparius larvae to 10, 50 and 100 micrograms l-1 4NP. Survival of the larvae was not affected by the tested concentrations, but the frequency of mentum deformities increased significantly (P < 0.01) after exposure to 4NP.     

Contaminated sediments and bioassay responses of three macroinvertebrates, the midge larva Chironomus riparius, the water louse Asellus aquaticus and the mayfly nymph Ephoron virgo

Bioassays are widely used to estimate ecological risks of contaminated sediments. We compared the results of three whole sediment bioassays, using the midge larva Chironomus riparius, the water louse Asellus aquaticus, and the mayfly nymph Ephoron virgo. We used sediments from sixteen locations in the Dutch Rhine-Meuse Delta that differed in level of contamination. Previously developed protocols for each bioassay were followed, which differed in sediment pretreatment, replication, and food availability. The Chironomus bioassay was conducted in situ, whereas the other two were conducted in the laboratory. The measured endpoints, survival and growth, were related to contaminant levels in the sediment and to food quantity in water and sediment.

Only the response of A. aquaticus in the bioassay was correlated with sediment contamination. Food availability in overlying water was much more important for C. riparius and E. virgo, thereby masking potential sediment contaminant effects. We conclude that growth of A. aquaticus was depressed by sediment contamination, whereas growth of E. virgo and C. riparius was stimulated by seston food quantity. We discuss that the trophic state of the ecosystem largely affects the ecological risks of contaminated sediments.     

Biological responses of midge (Chironomus riparius) and lamprey (Lampetra fluviatilis) larvae in ecotoxicity assessment of PCDD/F-, PCB- and Hg-contaminated river sediments

Environmental Science and Pollution Research - We evaluated the utility of chironomid and lamprey larval responses in ecotoxicity assessment of polychlorinated dibenzo-p-dioxins, dibenzofurans...     

The influence of salinity on metal uptake and effects in the midge Chironomus maddeni

The influence of different porewater salinities (up to 12 g/L) on the toxicity and bioaccumulation of copper, zinc and lead from metal-spiked sediments was assessed using the midge, Chironomus maddeni. Survival of the larvae was significantly reduced at a porewater salinity of 12 g/L, but no effects were observed at 4 or 8 g/L. Both growth and survival of C. maddeni were reduced after exposure to salt/metal spiked sediments as compared to those exposed to sediments spiked with metals or salt alone. Increased salinity resulted in increased bioaccumulation of copper and zinc, but decreased bioaccumulation of lead. The observed patterns of bioaccumulation were not entirely explained by the modelled free ion activities of the metals, indicating that factors such as osmotic stress, consumption of metal-contaminated sediments or metal interactions may have been important as well. These results highlight the need to consider the influence of existing or potential salinization when undertaking hazard assessments of freshwater systems impacted by contaminants such as trace metals.     

Environmental chemicals with known endocrine potential affect yolk protein content in the aquatic insect Chironomus riparius

The use of vitellogenesis as a marker for possible effects of endocrine disrupting agents on insects was tested in the aquatic midge Chironomus riparius. As test substances the synthetic ecdysoid tebufenozide, and the endocrine disruptors bisphenol a and 4-n-nonylphenol were applied in a semi-static manner. The yolk protein contents of freshly emerged (24 h) male and female midges were determined by an ELISA procedure. In males, where always low amounts of immunoreactivity were apparent, yolk concentrations were lowered by 10% after a 80 microg/l tebufenozide treatment, and by 20-25% after exposition to bisphenol a at concentrations of 1, 100, and 3,000 microg/l. 4-n-nonylphenol contamination caused an inverted dose-response curve. At low test concentrations (1.9-30 microg/l) reduced yolk immunoreactivity occurred, while at medium concentrations (120 and 500 microg/l) no significant effects were observable. In the most highly contaminated group (2,000 microg/l) yolk protein immunoreactivity was elevated to 107% of the control. Female yolk protein contents were affected only in the 3,000 microg bisphenol a/l contaminated group, where yolk immunoreactivity was reduced by ca. 10% compared to the control.     

In situ bioassays with Chironomus riparius larvae to biomonitor metal pollution in rivers and to evaluate the efficiency of restoration measures in mine areas

In this study we evaluate the ability of an in situ bioassay with Chironomus riparius larvae, using larval development and growth as endpoints, to biomonitor water quality and to assess the biological recovery of metal contaminated freshwater ecosystems of mine areas that are subject of restoration measures. The bioassay was carried out in streams located near an abandoned goldmine in North Portugal, throughout an environmental rehabilitation of the mine (2002-2004). During this period, a decrease in the inhibition of larval growth in the metal contaminated stream was observed. The bioassay was also performed in streams located near an active tungsten mine in Central Portugal. Larval growth and development were highly inhibited in the stream that receives acid drainage from the tungsten mine and treated water from the AMD treatment station. The results indicate that the bioassay can be used to evaluate the efficiency of environmental restoration measures in mining areas.     

Exposure of Chironomus riparius larvae to uranium: effects on survival, development time, growth, and mouthpart deformities

Among non-biologically essential metals, data concerning uranium effects on freshwater benthic macroinvertebrates are scarce. The effects of uranium on survival, development time, growth and mouthpart deformities of midge Chironomus riparius were investigated. A 10-day static laboratory bioassay was performed exposing first instar larvae to artificial sediment spiked with four concentrations of uranium (2.97; 6.07; 11.44; 23.84 microg U g(-1) dry wt). As uranium was released from the sediment to the overlying water during this bioassay, both the sediment and the water column act as contamination pathways in giving rise to the observed effects. Significant negative effects on survival, development time, and growth were detected at 6.07, 6.07 and 2.97 microg U g(-1) dry wt, respectively. An LC20 of 2.49 microg U g(-1) dry wt (95% CI=1.48-4.27), and an LC50 of 5.30 microg U g(-1) dry wt (95% CI=3.94-7.25) were estimated. With respect to effects of uranium on larvae mouthpart deformities, we found that the lower the concentrations, the higher the deformity rates. These results highlight the potential impact of uranium at population level in environmentally realistic concentrations.     

Variation in ozone sensitivity among clones of Betula pendula and Betula pubescens

Forty clones of Betula pendula and 6 clones of Betula pubescens, originating from southern and central Finland, were ranked in order of ozone sensitivity according to visible injuries, growth and leaf senescense under low ozone exposure. The plants were fumigated in natural climatic conditions using an open-air exposure system during two growing seasons. Control plants were grown under ambient air, and the elevated-ozone exposures were 1.6x the ambient in 1994 and 1.7x the ambient in 1995. The differences in ozone sensitivity among clones were large. Ozone tolerance was related to thicker leaves and higher stomatal density as compared to sensitive clones. Ultrastructural ozone-induced symptoms were found in chloroplasts of sensitive clones. Increased number of visibly injured leaves on fumigated plants was correlated with reduced leaf formation, foliage area, shoot dry wt and number of stomata, and increased yellowing of leaves. The results suggest that a considerable proportion of birch trees, showing high sensitivity to ozone, are at risk if ambient ozone exposures increase.     

Exposure of Chironomus riparius larvae (diptera) to lead, mercury and beta-sitosterol: effects on mouthpart deformation and moulting

Mouthpart deformation in chironomid larvae is induced by exposure to chemical contaminants and is becoming an established bio-indicator in sediment assessment programmes. However, concentration-response relationships with causal agents have only been established occasionally and with varying success. In this laboratory study, instar II and III larvae were exposed to sub-lethal concentrations of lead, mercury and beta-sitosterol. A significant deformation response was induced in the pecten with lead and mercury. Deformation frequencies of the mentum after metal exposures were not significantly different from the control. Moulting was retarded by both metals and was well correlated with mouthpart deformation. The beta-sitosterol is an endocrine disruptor, which was used to test the hypothetical cause-effect relation between disruption of ecdyson functioning and chironomid deformation. In the present study, exposure to sublethal concentrations of beta-sitosterol did not result in any effect on deformation or moulting. As such, the proposed hypothesis of endocrine disruptors as primary causal agents of chironomid deformation could not be substantiated. Acetone, which was used as a solvent to apply beta-sitosterol caused a significant increase of mentum deformation. The ground filtration paper used as substrate seemed to induce deformities as well. Substrate contamination, acetone and (especially) inbreeding were most probably responsible for the high deformation frequencies in the control conditions.     

Short-term exposure to sub-lethal doses of lindane affects developmental parameters in Chironomus riparius Meigen, but has no effect on larval glutathione-S-transferase activity

Chironomus riparius Meigen were exposed to 0, 0.01, 0.1, 0.5, 0.75 and 1.0 ppm lindane for 48 h as fourth instar larvae. Exposure had no effect on glutathione-S-transferase (GST) activity in larvae snap-frozen immediately following exposure. In contrast, exposure had longer-term consequences affecting developmental parameters. Concentrations above 0.5 ppm lindane affected larval behaviour, reduced adult body size and fecundity and delayed emergence times. The lack of significant change in GST activity when life history characters were affected by high concentrations of lindane, suggests that in C. riparius, GST is not a sensitive biomarker of pesticide exposure or effect.     

Metallothionein modulation in relation to cadmium bioaccumulation and age-dependent sensitivity of <Emphasis Type="Italic">Chironomus riparius</Emphasis> larvae

The goal of this study was to contribute to understanding of the mechanisms behind sensitivity differences between early and late instar larvae of Chironomus riparius and to address the influence of the differences in standard testing approaches on the toxicity evaluation. A 10-day contact sediment toxicity test was carried out to assess sensitivity to cadmium exposure in relation to different age and laboratory culture line origin of test organisms. Chironomid larvae of early (OECD 218 method) and late instar (US-EPA600/R-99/064 method) differed substantially in sensitivity of traditional endpoints (OECD: LOEC 50 and 10 μg Cd/g dry weight (dw); US-EPA: LOEC?>?1000 and 100 μg Cd/g dw for survival and growth, respectively). Bioaccumulated cadmium and metallothioneins (MTs) concentrations were analyzed to investigate the role of MTs in reduced sensitivity to cadmium in late instar larvae. Metallothioneins were induced after treatment to greater Cd concentrations, but their levels in relation to cadmium body burdens did not fully explain low sensitivity of late instars to cadmium, which indicates some other effective way of detoxification in late instars. This study brings new information related to the role of MTs in age-dependent toxicant sensitivity and discusses the implications of divergence in data generated by chironomid sediment toxicity tests by standardized methods using different instars.     

Use of whole-body and subcellular Cu residues of Lumbriculus variegatus to predict waterborne Cu toxicity to both L. variegatus and Chironomus riparius in fresh water

We tested the use of whole-body and subcellular Cu residues (biologically-active (BAM) and inactive compartments (BIM)), of the oligochaete Lumbriculus variegatus to predict Cu toxicity in fresh water. The critical whole-body residue associated with 50% mortality (CBR₅₀) was constant (38.2-55.6 μg g⁻¹ fresh wt.) across water hardness (38-117 mg L⁻¹ as CaCO₃) and exposure times during the chronic exposure. The critical subcellular residue (CSR₅₀) in metal-rich granules (part of BIM) associated with 50% mortality was approximately 5 μg g⁻¹ fresh wt., indicating that Cu bioavailability is correlated with toxicity:subcellular residue is a better predictor of Cu toxicity than whole-body residue. There was a strong correlation between the whole-body residue of L. variegatus (biomonitor) and survival of Chironomus riparius (relatively sensitive species) in a hard water Cu co-exposure. The CBR₅₀ in L. variegatus for predicting mortality of C. riparius was 29.1-45.7 μg g⁻¹ fresh wt., which was consistent within the experimental period; therefore use of Cu residue in an accumulator species to predict bioavailability of Cu to a sensitive species is a promising approach.     

Monitoring of genetically modified Escherichia coli in laboratory wastewater

Containment of genetically modified (GM) microorganisms such as Escherichia coli is a legal requirement to protect the environment from an unintended release and to avoid horizontal gene transfer (HGT) of recombinant DNA to native bacteria. In this study, we sampled the laboratory wastewater (LWW) at a large Swiss university from three sources over 2 years and cultured ampicillin-resistant, presumptive GM E. coli. From a total of 285 samples, 127 contained presumptive GM E. coli (45%) at a mean concentration of 2.8 × 10² CFU/ml. Plasmid DNA of 11 unique clones was partially or entirely sequenced. All consisted of cloning vectors harboring research-specific inserts. To estimate the chance of HGT between GM E. coli and native bacteria in LWW, we identified taxa representative for the bacterial community in LWW using 16S rRNA amplicon sequencing and measured conjugation frequencies of E. coli with five LWW isolates. At optimal conjugation conditions, frequencies were between 3.4 × 10⁻³ and 2.4 × 10⁻⁵. Given the absence of transferable broad-host range plasmids and suboptimal conjugation conditions in the LWW system, we conclude that the chance of HGT is relatively low. Still, this study shows that the implementation of robust containment measures is key to avoid the escape of GM microorganisms.

     

Functional and structural rearrangements of salivary gland polytene chromosomes of Chironomus riparius Mg. (Diptera,Chironomidae) in response to freshly neutralized aluminium

Although recent work has shown that environmentally relevant concentrations of freshly neutralized aluminium (AI) are bioavailable and toxic to freshwater invertebrates, the genotoxicity of Al has not been examined. Here we show that freshly neutralized Al affects structure and function of the salivary gland polytene chromosomes of the ubiquitous chironomid larva Chironomus riparius over three generations. Exposure to 500 microg l-1 added Al for 24-25 days resulted in a significantly higher frequency of numerous somatic aberrations, while no structural aberrations were found in F1 controls and few in the second and third generation. Aberrations also included deletions of sections of chromosome G of C. riparius larvae as well as deletions of one or more Balbiani rings. Changes in functional activity included decreased activity of the Balbiani rings (BR), and an increase in the number of decondensed centromeres. The activity of the nucleolar organizer (NOR) significantly decreased in F1 chironomids exposed to Al, while in the F2 and F3 generations the NOR showed normal (high) activity. First generation chironomids were generally more susceptible to Al although no clear evidence of tolerance was apparent over three generations. The possible use of alterations in chironomid polytene chromosomes as biomarkers of trace metal pollution is discussed.