Chironomus Xanthus Research Articles

An evaluation of Allonais inaequalis Stephenson, 1911 (Oligochaeta: Naididae) as a toxicity test organism

Oligochaeta are suitable for assessing the effects of contaminants on the environment, but in Brazil, few studies have been developed for the use of native species in toxicity bioassays. Given the increasing interest in this research area by the environmental agencies for the usage of species of aquatic Oligochaeta in ecotoxicological studies, in addition to the need for greater biological knowledge of native species, the present work aimed to establish a protocol to be used in ecotoxicological bioassays using the native species Allonais inaequalis. The Naididae A. inaequalis (Stephenson, 1911) was first described as Nais pectinata var. inaequalis Stephenson, 1911. The distribution of this species has been mentioned for several environments in Brazil. This paper presents results from tests on the sensitivity of the species to KCl (LC50) and to zinc chloride (LC50), the general characteristics of the species and laboratory breeding methods. Laboratory rearing, the collection and identification of the organism are easy when compared to other organisms such as Chironomus xanthus, a species commonly used in toxicity bioassays. It is expected that this work will assist in the use of A. inaequalis species for routine water monitoring protocols for tropical environments.

Flavonoids from Kalanchoe ×laetivirens Impair Survival and Immunity of Atta sexdens (Hymenoptera: Formicidae)

Background: The active secondary metabolites of various Kalanchoe species are promising for controlling insects and other agricultural pests. Objective: This study aimed to evaluate the mortality and immune responses of the leafcutter ant Atta sexdens (L.) after treatment with Kalanchoe ×laetivirens (Desc.) aqueous extract. The effects on an aquatic organism used as a bioindicator, Chironomus xanthus (Rempel), were also evaluated. Methods: Different K. ×laetivirens concentrations were prepared by dilution in honey to assess the toxicity and determine the median lethal concentration (LC50) for A. sexdens workers. The haemocytes of workers treated with a sublethal concentration (0.005 g/mL) were counted. Results: The plant extracts induced high mortality in A. sexdens workers. All concentrations, except 0.001 g/mL, increased mortality rates. The sublethal concentration negatively affected the immune system of A. sexdens workers 48 and 72 h after ingestion. The flavonoids 4″″-acetylsagittatin A, sagittatin A and the nucleosides uridine and adenosine were identified. The LC50 of the K. ×laetivirens (128.1 mg /L) was determined for the non-target organism, C. xanthus. Conclusion: These natural compounds can be used in future research to plan alternative control strategies for leafcutter ants in forests and other agroecosystems.

The combined effects of microbial insecticides and sodium chloride on the development and emergence of Chironomus xanthus.

Freshwater organisms are facing increasing salinity levels, not only due to natural environmental processes, but also human activities, which can cause several physiological adaptations to osmotic stress. Additionally, these organisms might also have to deal with contamination by microbial insecticides. Our main goal was to use Chironomus xanthus to assess the chronic effects of increasing the salinity and commercial formulations of the microbial insecticides based on Bacillus thuringiensis subs. kurstaki (Btk) and Beauveria bassiana (Bb) as active ingredients, respectively. A significant interaction of growth was observed between the biopesticide based on Bb and NaCl on the larvae of C. xanthus. Single exposure to NaCl and each one of the formulations demonstrated deleterious impacts not only on larval development, but also on the emergence success and emergence time of this nontarget insect, with potential consequences for freshwater ecosystems due to cascading effects. The chronic effects induced by both bioinsecticides show that these formulations can have environmental impacts on nontarget freshwater insects. © 2023 Society of Chemical Industry.

What does not kill it makes it stronger! The tolerance of the F1 larvae of Chironomus xanthus to a neonicotinoid insecticide formulation

Thiamethoxam (TMX) is a systemic neonicotinoid that acts as a partial agonist of the nicotinic acetylcholine receptors (nAChRs). However, target species have shown resistance to formulations based on such neonicotinoids, which can also be expected for non-target insects. This research aimed to study the effects of a formulation based on TMX [Cruiser® 350 FS (CRZ)] on the life traits of Chironomus xanthus filial generation (F1) and compare it with the parental generation (P). Environmentally relevant concentrations of CRZ significantly decreased larvae growth P generation , also slowing and decreasing their emergence. Larvae of the F1 generation were less sensitive than their parents, suggesting that the progeny were able to thrive and perform basic physiological functions better than the parental generation. Our results highlight that insect resistance to neonicotinoids may be associated with the better performance of the filial generation, which is related to the change in affinities of the active ingredient for the sub-units constituting the nAChRs subtypes of F1 organisms, inherited from P organisms that were able to survive and reproduce. Moreover, further studies using biochemical and omics tools should be performed to disentangle the specific changes occurring at the nAChRs throughout insect development.

The multigenerational effects of clothianidin on Chironomus xanthus: Larvae exposed to this acetylcholine super agonist show no clear resistance

Clothianidin (CLO) is an insecticide belonging to the second-generation class of neonicotinoids. In this study, we evaluated how CLO affects the survival and the complete life cycle of the tropical insect Chironomus xanthus, a non-target species, considering the Parental (P) and Filial (F1) generations. We found a 48 h-lethal concentration (LC50) of CLO of 3.78 µg/L. The lowest observed effect concentrations (LOECs) were: i) for body growth and head capsule width in P generation = 47.3 ng/L CLO; ii) for body growth and head capsule width in F1 generation larvae = 80 and 36.4 ng/L CLO, respectively; iii) for cumulative emergence it was 80 ng/L CLO in the P generation, while there was no significant difference in the F1 generation; iv) for total developmental time for males and females = 61.53 ng/L in P generation; v) in the F1 generation, the LOEC was determined to be 36.4 ng/L for males and 80 ng/L for females; vi) The number of total hatched eggs and total hatched eggs/female had LOECs of 36.4 ng/L CLO for both generations. Our study reveals that environmentally relevant concentrations of the CLO-based insecticide are highly toxic to C. xanthus. It also shows that the F1 generation, resulting from parents exposed to CLO was not clearly resistant to the insecticide. This fact might be explained by the different effects observed for males and females of F1 generation. Understanding the sub-types of acetylcholine receptors present on target and non-target insect species and toxicological effects of neonicotinoids seems to be desirable for the insecticide industry to deal with insect pests and the environmental protection of non-target organisms.

Is there a common mechanism of neonicotinoid resistance among insects? Preliminary results show that F1 larvae of pre-exposed Chironomus xanthus are more tolerant to imidacloprid

Imidacloprid (IMI) belongs to the group of the first generation of neonicotinoids, known for their toxic effects to non-target invertebrate species. Neonicotinoids have a specific mode of action that mimics acetylcholine, impairing the cholinergic neurotransmission, ultimately affecting the survival of organisms. This research aimed to determine the effects of a commercial formulation based on IMI on survival, and life traits parameters to larvae of parental (P) and F1 generations of the non-target insect Chironomus xanthus. C. xanthus is a bioindicator species of environmental change of great ecological relevance in tropical freshwater systems. The 48-h LC50 of IMI was 23.72 µg/L. Environmentally relevant concentrations of IMI impaired the development of larvae, delayed emergence and diminished the fecundity and fertility rates. However, a physiological adaptation of C. xanthus larvae from the parental generation to the filial generation can be suggested. This finding seems to be a key factor determining the resistance to neonicotinoids in pest species. Further biochemical studies should be carried out to unravel possible physiological adaptations focused on detoxification processes and variability of nicotinic receptors that allow the increase of resistance over generations of insect species of interest.

Lethal and sublethal effects of the saline stressor sodium chloride on Chironomus xanthus and Girardia tigrina.

Salinization in freshwaters is gradually increasing as a result of human activities and climatic changes. Higher salt content causes stress for freshwater organisms. Sodium chloride (NaCl) is among the most frequently occurring salts in freshwater ecosystems. The objective of the present study was to investigate the lethal and sublethal effects of NaCl on freshwater ecosystems, using as test organism the dipteran Chironomus xanthus and the planarian Girardia tigrina. Acute tests showed that C. xanthus was more sensitive (48-h LC50 (median lethal concentration) of 2.97gNaClL-1) than G. tigrina (48-h LC50 of 7.77gNaClL-1). C. xanthus larvae growth rate (larvae length and head capsule width) was significantly reduced under exposure to concentrations as low as 0.19gL-1 NaCl and higher. A delay in the emergence time (EmT50) was also demonstrated for the same concentration. Sublethal NaCl effects in G. tigrina included feeding inhibition (LOEC (lowest observed effect concentration) of 0.4gL-1), reduced locomotion (LOEC = 0.2gL-1), and 24-48-h blastema regeneration (LOEC = 0.2gL-1 and 0.1gL-1, respectively). The results demonstrated the toxicity of NaCl to C. xanthus and G. tigrina including sublethal effects that can result in negative consequences for populations in natural freshwaters under salinization.

Effects of lethal and sublethal concentrations of peracetic acid and active chlorine of calcium hypochlorite on Chironomus xanthus

Freshwater ecosystems are vulnerable to residual concentrations of chemical agents from anthropogenic activities, and the real impacts of such compounds can only be evaluated accurately using ecotoxicological tests. The assessment of ecotoxicological effects of peracetic acid (PAA) and the active chlorine of calcium hypochlorite (Ca(ClO)2) on the insect Chironomus xanthus Meigen (Diptera: Chironomidae) is highly relevant as there are few reports on its effects in fresh water ecosystems. To our best knowledge, this is the first study to assess the chronic toxicity of the compounds to C. xanthus. The toxicity bioassays for C. xanthus included the acute effect (CL50) and the chronic effects based on body length, head width, and cumulative emergence. The results obtained in the acute effect tests indicated that the active chlorine of Ca(ClO)2 is 14 fold more toxic than PAA to C. xanthus. In sublethal evaluations, the active chlorine of Ca(ClO)2 presented higher toxicity than PAA in terms of percentage emergence, body development, and head width. In general, the results showed lower PAA toxicity relative to the active chlorine of Ca(ClO)2, demonstrating that PAA is a promising substitute for chlorinated disinfectants. In addition, the study facilitates the establishment of reference values for the safe release of effluents treated with PAA into water bodies.

Toxicity of microbial insecticides toward the non-target freshwater insect Chironomus xanthus.

Commercial formulations based on Bacillus thuringiensis subs. kurstaki (Btk) and Beauveria bassiana (Bb) are commonly used microbial insecticides in Brazil and other tropical regions. However, and despite being considered environmentally friendly, their use generates concerns regarding possible adverse ecological effects in freshwater ecosystems. Here, we evaluate the effects of these bioinsecticides on the tropical aquatic dipteran Chironomus xanthus under laboratory conditions. After laboratory exposures to these compounds 48-h median lethal concentration (LC50 ) values of 1534 μg a.i./L for Btk and of 6.35 μg a.i./L for Bb were estimated. Chronic assays revealed different sublethal effects: Btk-based bioinsecticide exposure reduced C. xanthus growth [lowest observed effect concentration (LOEC) was 126 μg a.i./L for head width], decreased emergence rate (LOEC = 8 μg a.i./L) and increased immunological response (LOEC = 50 μg a.i./L) measured as total hemocyte count in larvae hemolymph. Exposure to low concentrations of Bb-based insecticide also reduced C. xanthus growth (LOEC = 0.07 μg a.i./L for larvae body length measurements), and emergence rate (LOEC = 0.28 μg a.i./L), despite no clear effects on the total hemocyte counts. Our results suggest that low concentrations of Btk and Bb bioinsecticides are toxic to C. xanthus. Given their widespread use and occurrence in tropical freshwater systems, research is needed to evaluate the potential effects of these compounds concerning natural freshwater insect communities and ecosystem functioning. © 2019 Society of Chemical Industry.

The direct effects of a tropical natural humic substance to three aquatic species and its influence on their sensitivity to copper.

Few studies have been conducted so far into the effects of humic substances (HS) on aquatic organisms and their influence on the toxicity of chemical pollutants in the tropics. The aim of the present study was therefore to evaluate the direct effects of locally-derived tropical natural HS on the cladoceran Daphnia similis, the midge Chironomus xanthus and the fish Danio rerio. The influence of a HS concentration series on the acute toxicity of copper to these organisms was also assessed through laboratory toxicity testing. The HS did not exert direct acute effects on the test organisms, but long-term exposure to higher HS concentrations provoked a stress response (increase in feces production) to D. rerio and exerted effects on chironomid adult emergence and sex ratio. The biotic ligand model proved to be a useful tool in converting total copper concentrations to the appropriate bio-available fraction to which tropical aquatic organisms are exposed.

Effects of a Thiamethoxam-Based Insecticide on the Life History of Chironomus xanthus

Thiamethoxam (TMX) is one of the most widely used neonicotinoid insecticides for pest control in agricultural crops. However, information on its acute and chronic toxicity for freshwater non-target insects is still limited. We aimed to evaluate the lethal and sub-lethal effects of TMX-based insecticide on Chironomus xanthus using laboratory ecotoxicological assays. Besides survival, 28-day partial life-cycle tests assessed the effects of chronic exposures on larval growth and emergence of midges. The estimated 48-h LC50 was 32 μg a. i./L of TMX. Chronic tests revealed that chironomids exposed to sub-lethal concentrations of TMX showed reduced larval growth (LOEC = 0.4 and 1.6 μg a.i/L of TMX for head capsule width and growth rate, respectively) and emergence rate (LOEC of 1.6 μg a.i/L of TMX). This study provides important ecotoxicological data concerning effects of TMX-based insecticides on tropical ecosystems and reveals that environmentally relevant concentrations of TMX can adversely affect chironomids populations. Our study also validates C. xanthus as a sensitive species to monitor the ecological integrity of tropical aquatic systems near intensive agricultural areas and to help understand the effects caused by TMX on freshwater insects.

Low Concentrations of Glyphosate-Based Herbicide Affects the Development of Chironomus xanthus

Glyphosate is an herbicide commonly used worldwide for weed control and generally applied as part of a formulated product, such as Roundup. Contamination of surface water by glyphosate-based herbicides can cause deleterious effects in organisms, mainly in aquatic systems near to intensive agricultural areas (e.g., transgenic soybean crops). Given the lack of toxicological information concerning effects of glyphosate-based herbicides on tropical aquatic ecosystems, we aimed to evaluate the lethal and sub-lethal effects of Roundup Original® on the dipteran Chironomus xanthus. The endpoints evaluated included survival, growth, and emergence. The results showed that the 48 h LC50 for glyphosate to C. xanthus was 251.5 mg a.e./L. Larval growth of C. xanthus was reduced under glyphosate exposure (LOEC for body length = 12.06 mg/L; LOEC for head capsule width = 0.49 mg/L). No effects were observed in terms of cumulative percentage of imagoes emergence. However, low concentrations of glyphosate caused delayed emergence of females (at 1.53 mg/L) and induced fast emergence of males (at 0.49 mg/L), compared to control treatment. The deleterious effects of environmental relevant concentrations of glyphosate (0.7 mg/L) observed in terms of C. xanthus growth and development suggest that glyphosate-based herbicides can have negative consequences for aquatic non-target invertebrates such as Chironomus. Multigerational assays are needed to assess the long term effects of glyphosate on C. xanthus populations. Finally, our study adds ecotoxicological data on the effects of glyphosate-based herbicides on tropical freshwater invertebrates.

Toxicity on aquatic organisms exposed to secondary effluent disinfected with chlorine, peracetic acid, ozone and UV radiation.

The toxic potential of four disinfectant agents (chlorine, ozone, peracetic acid and UV radiation), used in the disinfection of urban wastewater, was evaluated with respect to four aquatic organisms. Disinfection assays were carried out with wastewater from the city of Araraquara (São Paulo State, Brazil), and subsequently, toxicity bioassays were applied in order to verify possible adverse effects to the cladocerans (Ceriodaphnia silvestrii and Daphnia similis), midge larvae Chironomus xanthus and fish (Danio rerio). Under the experimental conditions tested, all the disinfectants were capable of producing harmful effects on the test organisms, except for C. xanthus. The toxicity of the effluent to C. silvestrii was observed to increase significantly as a result of disinfection using 2.5mgL(-1) chlorine and 29.9mgL(-1) ozone. Ozonation and chlorination significantly affected the survival of D. similis and D. rerio, causing mortality of 60 to 100% in comparison to the non-disinfected effluent. In experiments with effluent treated with peracetic acid (PAA) and UV radiation, a statistically significant decrease in survival was only detected for D. rerio. This investigation suggested that the study of the ideal concentrations of disinfectants is a research need for ecologically safe options for the treatment of wastewater.

Assessment of the acute toxicity of eutrophic sediments after the addition of calcium nitrate (Ibirité reservoir, Minas Gerais-SE Brazil): initial laboratory experiments

This study evaluated the acute toxicity of sediment in a eutrophic reservoir after remediation with a calcium nitrate solution to retain phosphorus. The study involved microcosms of surface sediments and water from the sediment-water interface in the Ibirité reservoir. This reservoir, located in the vicinity of metropolitan Belo Horizonte (Minas Gerais, SE Brazil), is a water body that receives treated effluents from an oil refinery (REGAP-Petrobras), as well as high loads of untreated urban effluents from the city of Ibirité and surrounding areas and industrial effluents from a major industrial park. Incubation times of the treatment experiments were: t = 0, t = 5, t = 10, t = 25, t = 50, t = 85 and t = 135 days. One control microcosm and three treated microcosms were analysed in each time interval. Acute toxicity of water samples was assessed with Ceriodaphnia silvestrii Daday, 1902 and that of bulk sediment samples with Chironomus xanthus Rempel, 1939. Toxicity tests were carried out concomitantly with chemical analyses of dissolved inorganic nitrogen species (ammonia, nitrate and nitrite), sulfate and metals in the water samples of the microcosms. Acid volatile sulfides (AVS), simultaneously extracted metal (SEM) and potentially bioavailable metal were analyzed in bulk sediment samples. Neither of the tested organisms showed toxicity in the control microcosm samples. The water column of the treated microcosm showed toxicity to C. silvestrii, starting at t = 10 days, while the sediment pore water toxicity started at t = 0 day. However, toxicity was found to decline from t = 85 days to t = 135 days. Sediments showed toxicity to C. xanthus during the entire experiment, except at the longest incubation time (t = 135 days). The overall results indicate that nitrate, which reached concentrations exceeding 1,200 mg N-NO3- L-1 in the sediment pore water of the treated microcosms, was most probably responsible for the toxicity of the samples. Although the calcium nitrate technology proved effective in retaining phosphorus, promoting sediment oxidation via denitrification, from the ecotoxicological standpoint and under the experimental conditions of this study, the application of nitrate for remediation of the sediments in the Ibirité reservoir did not prove effective up to a period of 135 days of incubation. However, we presume that after longer periods of incubation, treated sediments may recover their ability to sustain a benthic community. More advanced experiments are planned involving longer incubation times, thus extending the denitrification process, which may lead to a higher phosphorus retention capacity and to more complete abatement of sediment toxicity.

Lethal effects of abamectin on the aquatic organisms Daphnia similis, Chironomus xanthus and Danio rerio

Abamectin is used as an acaricide and insecticide for fruits, vegetables and ornamental plants, as well as a parasiticide for animals. One of the major problems of applying pesticides to crops is the likelihood of contaminating aquatic ecosystems by drift or runoff. Therefore, toxicity tests in the laboratory are important tools to predict the effects of chemical substances in aquatic ecosystems. The aim of this study was to assess the potential hazards of abamectin to the freshwater biota and consequently the possible losses of ecological services in contaminated water bodies. For this purpose, we identified the toxicity of abamectin on daphnids, insects and fish. Abamectin was highly toxic, with an EC50 48h for Daphnia similis of 5.1ngL−1, LC50 96h for Chironomus xanthus of 2.67μgL−1 and LC50 48h for Danio rerio of 33μgL−1.

Laboratory measurements of biomarkers and individual performances in Chironomus xanthus to evaluate pesticide contamination of sediments in a river of southeastern Brazil

This study aimed at evaluating biomarkers, individual and population responses in the native Chironomus xanthus to assess the toxicity of pesticide-contaminated sediments from the Monjolinho River (Southeast Brazil). We measured cholinesterase (ChE) and glutathione S-transferase activities (GST), as biomarkers and survival, individual growth and adult emergence, as individual performances. There was no response of the ChE activity and a tendency to decreased GST activity in contaminated sites, but this was generally not statistically significant. Therefore, there was no association of the biomarker responses with exposure to sediment containing pesticides. In contrast, ash free dry mass was significantly increased and male emergence was decreased in C. xanthus exposed to the same sediments. In conclusion, the selected biomarkers were not sensitive and specific enough to detect and anticipate effects of pesticide contamination at the levels measured in the study area. Nevertheless, individual performances alterations pointed to potential pollution problems and possible ecological consequences.

Toxicity of ferric chloride sludge to aquatic organisms

Iron-rich sludge from a drinking water treatment plant (DWTP) was investigated regarding its toxicity to aquatic organisms and physical and chemical composition. In addition, the water quality of the receiving stream near the DWTP was evaluated. Experiments were carried out in August 1998, February 1999 and May 1999. Acute toxicity tests were carried out on a cladoceran (Daphnia similis), a midge (Chironomus xanthus) and a fish (Hyphessobrycon eques). Chronic tests were conducted only on D. similis. Acute sludge toxicity was not detected using any of the aquatic organisms, but chronic effects were observed upon the fecundity of D. similis. Although there were relatively few sample dates, the results suggested that the DWTP sludge had a negative effect on the receiving body as here was increased suspended matter, turbidity, conductivity, chemical oxygen demand (COD) and hardness in the water downstream of the DWTP effluent discharge. The ferric chloride sludge also exhibited high heavy metal concentrations revealing a further potential for pollution and harmful chronic effects on the aquatic biota when the sludge is disposed of without previous treatment.

Biochemical Biomarkers in Individual Larvae of Chironomus xanthus (Rempel, 1939) (Diptera, Chironomidae)

Biochemical biomarkers have shown to be useful toxicity tools in environmental assessment programmes. However, few studies involving freshwater invertebrate species have been performed and even fewer on tropical species. The aim of this paper was to evaluate cholinesterase (ChE) and glutathione-S-transferase (GST) activities in larvae of Chironomus xanthus (Chironomidae, Diptera) exposed to the organophosphate parathion (ethyl) and the metals copper and cadmium. Considering ChE, significant reduction in activity was observed following C. xanthus exposure to parathion and cadmium (48 and 96 h, respectively). Inhibition of ChE activity (IC50) related to parathion seemed to be a more consistent and sensitive effect criteria than the LC50. For GST, there was significant reduction in activity following parathion exposure (48 h). Copper did not elicit any change in ChE or GST activities in C. xanthus. The findings of this paper show the two biomarkers as promising tools for assessing exposure to contaminats in tropical regions.

Comparison Between Laboratory and “in Situ” Bioassays Using Chironomus xanthus in the Assessment of Sediment Toxicity in the Monjolinho River (São Carlos, SP)

“In situ” bioassays (96 h), using Chironomus xanthus, were conducted in October/03 and January/04, in 4 sampling stations at Monjolinho river. Laboratory bioassays were conducted using 60g of sediment and 240 ml of ASTM medium, for 96 h. Six organisms were added to each test-chamber and the toxicity evaluation was given by the mean survival. The “in situ” bioassay test chambers (PVC) were covered by a 300 μm mesh. The sediment of each sampling station was added to the chambers together with 10 organisms at IV instars. The laboratory bioassays showed survivals ranging between 0% (Federal, Oct/03) and 100% (Federal, Jan/04). The results “in situ” bioassays showed high mortality (between 84% and 97% in Oct/03 and between 60% and 70% in Jan/04). However, the results did not show, necessarily, that the sediments were toxic or not, because of the high mortality that occurred in the river ́s spring (over 60%), which may be associated to other factors, like screen saturation with suspended matter and different sediment textures. The presence of indigenous organisms may have influenced the results, due the competition and predation in the chamber. Thus, we can infer that the protocol for “in situ” bioassays using C. xanthus must be improved in order to minimize the influence of confounding factors on the tests results.

Short‐term sublethal (sediment and aquatic roots of floating macrophytes) assays with a tropical chironomid based on postexposure feeding and biomarkers

This study proposes assays with a freshwater chironomid, Chironomus xanthus, distributed over South America, based on subindividual (acetylcholinesterase activity) and individual (survival and postexposure feeding) level endpoints. Sediment and aquatic-rooted floating macrophyte assays were developed, due to the importance of both substrates for toxicant exposure in subtropical/tropical environments. Assays were evaluated under realistic exposure scenarios by simulating a runoff over an agricultural field dosed with deltamethrin. In situ assays were performed within microcosms to discriminate the effects of deltamethrin from those of additional potential stress factors (organism handling and caging, microcosms, use of aquatic roots, and runoff per se). A laboratory sediment assay based on feeding was conducted with samples from the microcosms. In all assays, both sublethal endpoints were responsive to deltamethrin and more sensitive than survival. Survival and feeding were more sensitive in situ than in the laboratory. In the in situ sediment assays, both sublethal endpoints were within a similar range of sensitivity; they were significantly inhibited as of the lowest Decis dose, from approximately 20 to 70%. In situ feeding was more sensitive in the sediment than in the macrophyte assay, where it was inhibited significantly only at the two highest Decis doses (up to approximately 60%). Larval performance was not influenced significantly by any of the other potential stress factors.