Suspended particles only marginally reduce pyrethroid toxicity to the freshwater invertebrate Gammarus pulex (L.) during pulse exposure.

Current ecotoxicological research on particle-associated pyrethroids in freshwater systems focuses almost exclusively on sediment-exposure scenarios and sediment-dwelling macroinvertebrates. We studied how suspended particles influence acute effects of lambda-cyhalothrin and bifenthrin on the epibenthic freshwater amphipod Gammarus pulex (L.) using brief pulse exposures followed by a 144 h post exposure recovery phase. Humic acid (HA) and the clay mineral montmorillonite (MM) were used as model sorbents in environmentally realistic concentrations (5, 25 and 125 mg L(-1)). Mortality of G. pulex was recorded during the post exposure recovery phase and locomotor behavior was measured during exposure to lambda-cyhalothrin. We found that HA in concentrations ≥25 mg L(-1) adsorbed the majority of pyrethroids but only reduced mortality of G. pulex up to a factor of four compared to pyrethroid-only treatments. MM suspensions adsorbed a variable fraction of pyrethroids (10% for bifenthrin and 70% for lambda-cyhalothrin) but did not significantly change the concentration-response relationship compared to pure pyrethroid treatments. Behavioral responses and immobilisation rate of G. pulex were reduced in the presence of HA, whereas behavioral responses and immobilisation rate were increased in the presence of MM. This indicates that G. pulex was capable of sensing the bioavailable fraction of lambda-cyhalothrin. Our results imply that suspended particles reduce to only a limited extent the toxicity of pyrethroids to G. pulex and that passive uptake of pyrethroids can be significant even when pyrethroids are adsorbed to suspended particles.

Local physical habitat quality cloud the effect of predicted pesticide runoff from agricultural land in Danish streams.

The combination of intensive agricultural activities and the close connectivity between land and stream emphasise the potential risk of pesticide exposure in Danish streams. Benthic macroinvertebrates are applied in the assessment of stream ecological status, and some sensitive species have been shown to respond strongly to brief pulses of pesticide contamination. In this study we investigate the impact of agriculturally derived pesticides on stream macroinvertebrate communities in Denmark. As a measure of toxic pressure we apply the Runoff Potential. We investigated a total of 212 streams. These were grouped into distinct classes according to the magnitude of pesticide contamination in the period from 2003-2006. A total of 24 different macroinvertebrate indices were applied to detect effects of pesticide runoff (e.g. the SPEAR-index and the number of EPT taxa). We found high predicted pesticide runoff in 39% of the streams, but we found no significant effect of predicted pesticide exposure on stream macroinvertebrate indices. We, additionally, examined the influence of a series of environmental parameters ranging from site scale to catchment scale on the macroinvertebrate community. Relative proportions of gravel, sand and silt in bed sediments explained most of the variation in macroinvertebrate indices as well as the upstream riparian habitat quality. We suggest that the Runoff Potential model overestimate pesticide runoff contamination in Danish streams due the presence of buffer strips enforced by Danish legislation. When pesticide runoff contamination is low to moderate, poor physical properties (indirectly related to agricultural activity) are the main impediment for the ecological quality of Danish streams.

Vulnerability of Aquatic Insect Species to Insecticides, Depending on Their Flight Period and Adult Life Span.

Effects of insecticides on terrestrial adult life stages of otherwise aquatic insects, such as mayflies (Ephemeroptera), stoneflies (Plecoptera), and caddisflies (Trichoptera), are largely unknown. In the present study, a risk model was used to pinpoint the species most likely to experience effects due to spray drift exposure during the adult life stage. Using data from an earlier case study with lambda-cyhalothrin, 6 species with different life cycle traits were used to explore how life cycle characteristics may influence vulnerability. In addition, we performed a generic calculation of the potential effect on the terrestrial life stages of 53 species (including 47 species with unknown sensitivity). Our approach incorporated temporal and spatial distribution of both the insect and the insecticide, creating different exposure conditions among species due to variation in the relative proportion of the populations present at the time of insecticide spraying. The Ephemeroptera species represented were least vulnerable due to their extremely short adult life span and relatively short flight period. Based on their life cycle characteristics, Plecoptera and Trichoptera species were more vulnerable. These vulnerable species segregated into 2 distinct groups; one with a long adult life span to emergent period ratio and another with a high overlap between emergent period and spraying season. We therefore recommend that future ecotoxicological tests be done on species with these life cycle characteristics. Environ Toxicol Chem 2021;40:1778-1787. © 2021 SETAC.

Repeated insecticide pulses increase harmful effects on stream macroinvertebrate biodiversity and function.

We exposed twelve mesocosm stream channels and four instream channels to one, two, and four pulses of the insecticide lambda-cyhalothrin (0.1 µg L-1) applied at two day intervals, each pulse lasting 90 min. Unexposed controls were included. We monitored macroinvertebrate taxonomic composition in the channels and in deployed leaf packs one day before and 29 days after the first exposure. Further, we measured drift in and out of the channels and leaf litter decomposition. Lambda-cyhalothrin exposures induced significantly increased drift in both experiments especially for Gammarus pulex, Amphinemura standfussi, and Leuctra spp. Macroinvertebrate taxonomic composition increasingly changed with increasing number of lambda-cyhalothrin exposures being most pronounced in the mesocosm channels. Further, leaf decomposition significantly decreased with increasing number of exposures in the mesocosm channels. Our study showed that species with predicted highest sensitivity to lambda-cyhalothrin were primary drivers of significant changes in taxonomic composition lasting for at least one month despite continuous recolonization of exposed channels from upstream parts of the natural stream and from the water inlet in the mesocosm channels. The overall results highlight the importance of sequential exposures to insecticides for understanding the full impact of insecticides on macroinvertebrates at the community level in streams.

Effects of low flow and co-occurring stressors on structural and functional characteristics of the benthic biofilm in small streams.

Low flow and co-occurring stress is a more and more frequent phenomenon these years in small agricultural streams as a consequence of climate change. In the present study we explored short and longer term structural responses of the stream benthic algae community and biofilm metabolism to multiple stress in small streams applying a semi-experimental approach. We hypothesized that i) a reduction in flow in combination with secondary stress (nutrients and sediments) have immediate effects on the benthic algae community in terms of biomass (chlorophyll a, biovolume), taxonomic and trait (lifeform and size distribution) compositions as well as on metabolism (GPP and CR), and ii) that changes in the benthic algae community persist due to altered environmental settings but that functional redundancy among benthic algae species provides a high level of resilience in metabolism (GPP and CR). Overall, we found that stress imposed by nutrients was less pronounced than stress imposed by fine sediments under low flow, and that nutrient enrichment to some extent mitigated effects of fine sediments. Fine sediment deposition mediated a decline in the fraction of erect algae and/or algae with mucilage stalks but this did not happen under co-occurring stress from both sediments and nutrients. Additionally, fine sediment deposition mediated a decline in GPP of the biofilm, but again this did not happen under co-occurring stress from nutrients. We conclude that 1) the benthic algae community and biofilm metabolism displayed similar resilience to stress imposed by low flow and co-occurring stress from nutrients and sediments on a short and longer time scale and 2) as structure-function adaptations may occur at several trophic levels in the biofilm, more research is needed to explore mechanisms underlying mitigating effects of nutrients in response to sediment deposition under low flow.

Terrestrial adult stages of freshwater insects are sensitive to insecticides.

Terrestrial adult stages of freshwater insects may be exposed to pesticides by wind drift, over-spray, contact or feeding. However, studies addressing insecticide effects on freshwater invertebrates focus primarily on the impact of pesticides reaching the streams and potentially harming the aquatic juvenile stages. This is also reflected in the current risk assessment procedures, which do not include testing of adult freshwater insects. In order to assess the potential impact of insecticides on adult stages of freshwater insects, we exposed six common species to the insecticides Karate (lambda-cyhalothrin) and Confidor (imidacloprid). Dose-response relations were established, and LD50 estimates were compared to those of the honey bee, Apis mellifera L. (Hymenoptera: Apidae), which is the standard terrestrial test insect when pesticides are evaluated prior to commercial release. Generally, the tested species were more sensitive to the studied insecticides than the honey bee. In order to examine whether the sensitivity of adult stages of freshwater insects corresponds with the sensitivity of the juvenile stages of the same species, the ranking of the two life stages with respect to the toxicity of Karate was compared, revealing some correspondence, but also some dissimilarities. Our results strongly indicate that terrestrial adult stages of aquatic insects are not adequately protected by current risk assessment procedures.

Seasonal sensitivity of Gammarus pulex towards the pyrethroid cypermethrin.

The aquatic toxicity of insecticides like the pyrethroids have been discussed intensively over the recent years especially in relation to risk assessment and how seasonality may or may not affect the sensitivity of non-target organisms. To address this issue, the crustacean Gammarus pulex was collected once a month for 16 months and acclimated to 10 °C for four days before being exposed to a 90 min pulse of cypermethrin. In vitro cytochrome P450 activity, total lipid content, total protein content, and dry weight were measured in male and female gammarids from each sampling date and used along with the water temperature as variables for sensitivity prediction by Partial Least Squares (PLS) regression models. The 24 h EC50-values varied more than 30 fold across the sampling period from 0.21 ±â€¯0.05 µg L-1 (April 2015) to 6.60 ±â€¯3.46 µg L-1 (October 2015), indicating seasonal variances in the acute sensitivity of G. pulex towards cypermethrin. After 168 h of recovery this difference in EC50-values was reduced to seven-fold. In both male and female gammarids seasonal patterns were observed in the total lipid content and in vitro CYP P450 activity, which peaked in spring and fall, respectively. The current study shows the importance of reporting time of organism collection and experimental execution for risk assessment of pyrethroids as season is important for the acute sensitivity of G. pulex. We suggest prolonged acclimation times of sampled macroinvertebrates to constant laboratory conditions in order to even out possible seasonal differences in sensitivity.

Realistic pesticide exposure through water and food amplifies long-term effects in a Limnephilid caddisfly.

Pesticides are increasingly recognized as relevant stressors in stream ecosystems. Stream biota is exposed to pesticides with low water solubility, e.g. pyrethroid insecticides, via water, habitat, and food. However, long-term effects of simultaneous exposure pathways are unknown. In this context, we conducted a microcosm experiment with the caddisfly Anabolia nervosa exposing the larvae to the pyrethroid insecticide esfenvalerate (EFV) at 0.1 and 1.0µgL-1 via (i) water, (ii) food or a (iii) combination of water and food. Combined exposure through water and food significantly reduced emergence by 60% and significantly postponed emergence timing at the highest EFV level, whereas none of the single-phase exposures showed significant effects. Moreover, our study revealed that successfully emerged females from the highest biphasic treatment level were characterised by altered composition of storage lipids indicative of reduced energy reserves. Consequently, a realistic test scenario that represents simultaneous exposure of organisms and their food may reveal substantially increased long term effects of pyrethroids when compared with current ecological risk assessment applying only single phase exposure. We recommend that relevant concurrent exposure routes of pesticides should be considered in order to derive realistic regulatory acceptable concentrations of pesticides.

Multiple exposure routes of a pesticide exacerbate effects on a grazing mayfly.

Hydrophobic pesticides such as pyrethroid insecticides tend to occur in their soluble form mainly as transient pulses in streams. In addition, they are regularly detected in significant quantities adsorbed to stream sediments and other organic in-stream structures. Consequently, stream biota is likely subjected to pesticide exposure via multiple routes. In this study we aimed at investigating the influence of exposure routes for the pyrethroid insecticide lambda-cyhalothrin on the grazing mayfly Heptagenia sulphurea. Therefore, H. sulphurea was exposed to lambda-cyhalothrin via single- (water or biofilm) or biphasic exposure (water and biofilm) at environmentally realistic concentrations (0, 0.1, 1µgL(-1)) and exposure duration (2h) in a full factorial design (n=5). Mortality, moulting frequency, and biofilm accrual (proxy for feeding rate) were recorded subsequent to a 7 d post exposure period. Mortality significantly increased and moulting frequency significantly decreased with increasing concentrations of lambda-cyhalothrin in the water phase whereas exposure via biofilm prompted no significant effects on these endpoints (α=0.05). Effect predictions systematically underestimated and overestimated effects for mortality and moulting frequency, respectively. Similarly, mayfly feeding rate was significantly reduced by water phase exposure whereas pre-exposed biofilm did not significantly affect this variable. However, we found a significant but non-systematic interaction between water phase and biofilm exposure on mayfly feeding rate. Our results show that exposure to the same pesticide via multiple exposure routes may increase the magnitude of effects beyond the level predicted from single phase exposures which has clear implications for the aquatic risk assessment of hydrophobic pesticides. However, our results additionally reveal that interactions between pesticide exposure routes may vary between selected dependent variables. We emphasize that unravelling the underlying mechanisms causing these discrepancies in interactive effects between exposure routes is a major aspect that should receive further attention in future research.

Influence of rice field agrochemicals on the ecological status of a tropical stream.

Many tropical countries contain a high density of protected ecosystems, and these may often be bordered by intensive agricultural systems. We investigated the chemical and ecological status of a stream connecting an area with conventional rice production and a downstream protected nature reserve; Mata Redonda. Three sites were sampled: 1) an upstream control, 2) in the rice production area and 3) a downstream site in Mata Redonda. We sampled benthic macroinvertebrates and pesticides in water and sediments along with supporting physical and chemical data. Pesticide concentrations in water exceeded current safety thresholds at sites 2 and 3, especially during the rainy season, and sediment associated pesticide concentrations exceeded current safety thresholds in three of six samples. Importantly, the highest predicted pesticide toxicity in sediments was observed at site 3 in the Mata Redonda confirming that the nature reserve received critical levels of pesticide pollution from upstream sections. The currently used macroinvertebrate index in Costa Rica (BMWP-CR) and an adjusted version of the SPecies At Risk index (SPEAR) were not significantly correlated to any measure of anthropogenic stress, but the Average Score Per Taxon (ASPT) index was significantly correlated with the predicted pesticide toxicity (sumTUD.magna), oxygen concentrations and substrate composition. Our results suggest that pesticide pollution was likely involved in the impairment of the ecological status of the sampling sites, including site 3 in Mata Redonda. Based on our results, we give guidance to biomonitoring in Costa Rica and call for increased focus on pesticide transport from agricultural regions to protected areas.

Pyrethroid effects on freshwater invertebrates: a meta-analysis of pulse exposures.

Pyrethroids are widely used insecticides that may seriously harm aquatic organisms. Being strongly hydrophobic, pyrethroids in solution occur only in short pulses but may be retained in sediments for longer periods. Consequently, most studies consider the chronic exposure of sediment dwelling organisms. We collected data from 16 studies to determine effect thresholds for stream macroinvertebrates exposed to short pyrethroid pulses evaluating lethal and sublethal ecologically relevant endpoints. Dose-response models showed EC50 for lethality, functional and behavioural endpoints down to 1/100, 1/100 and 1/1000 of the 48 h LC50 for Daphnia magna, respectively. The results indicate that the overall sensitivity of stream macroinvertebrates to pyrethroids may be higher than previously believed. This review shows the relevance of incorporating data on sublethal endpoints and appropriate post-exposure observation periods in future studies. The current risk assessment procedures and the higher tier approach are discussed in the light of our results.

Pesticide impacts on predator-prey interactions across two levels of organisation.

In this study, we aimed to evaluate the effects of a short pulse exposure of the pyrethroid lambda-cyhalothrin (LC) on the predator and anti-predator behaviour of the same species; Gammarus pulex. Predator behaviour, at the level of the individual, was studied in indoor microcosms using video tracking equipment during simultaneous exposure of the predator (G. pulex) and its prey (Leuctra nigra) during 90 min exposure of 1, 6.6 or 62.1 ngL(-1) LC. During an initial 30 min of exposure, the predator and prey organisms were maintained physically separated, and the actual interaction was studied through the subsequent 60 min of exposure. The anti-predator behaviour of G. pulex (drift suppression in response to the presence of brown trout) was studied in outdoor stream channels during a 90 min pulse exposure to LC (7.4 or 79.5 ngL(-1)) with, or without, brown trout. Based on survival curves for L. nigra we found that the mortality rate for L. nigra significantly decreased during exposure to 6.6 and 62.1 ngL(-1) LC (P<0.05 and P<0.001, respectively). We found no significant effects suggesting that G. pulex was repelled by contaminated prey items (P>0.05). We found that the exposure of G. pulex to 7.4 and 79.5 ngL(-1) LC significantly increased drift (from ∼0% to ∼100% in both treatments; P<0.001) independent of the presence of brown trout (P<0.05). In other words, the natural anti-predator behaviour of G. pulex was overruled by the stress response to LC exposure increasing G. pulex predation risk from drift feeding brown trouts. Our results show that the anti-predator and predator behaviour of G. pulex were significantly changed during exposure to very low and environmentally realistic LC concentrations and exposure duration. The potential implications for the field scenario are discussed.

Stream habitat structure influences macroinvertebrate response to pesticides.

Agricultural pesticides continue to impair surface water ecosystems, although there are few assessments of interactions with other modifications such as fine sediment and physical alteration for flood drainage. We, therefore, surveyed pesticide contamination and macroinvertebrates in 14 streams along a gradient of expected pesticide exposure using a paired-reach approach to differentiate effects between physically modified and less modified sites. Apparent pesticides effects on the relative abundance of SPEcies At Risk (SPEAR) were increased at sites with degraded habitats primarily due to the absence of species with specific preferences for hard substrates. Our findings highlight the importance of physical habitat degradation in the assessment and mitigation of pesticide risk in agricultural streams.

Effects of a triazole fungicide and a pyrethroid insecticide on the decomposition of leaves in the presence or absence of macroinvertebrate shredders.

Previously, laboratory experiments have revealed that freely diluted azole fungicides potentiate the direct toxic effect of pyrethroid insecticides on Daphnia magna. More ecologically relevant exposure scenarios where pesticides are adsorbed have not been addressed. In this study we exposed beech leaves (Fagus sylvatica) to the azole fungicide propiconazole (50 or 500 µg L(-1)), the pyrethroid insecticide alpha-cypermethrin (0.1 or 1 µg L(-1)) or any combination of the two for 3h. Exposed leaves were transferred to aquaria with or without an assemblage of macroinvertebrate shredders, and we studied treatment effects on rates of microbial leaf decomposition, microbial biomass (using C:N ratio as a surrogate measure) and macroinvertebrate shredding activity during 26 days post-exposure. Microbial leaf decomposition rates were significantly reduced in the propiconazole treatments, and the reduction in microbial activity was significantly correlated with loss of microbial biomass (increased C:N ratio). Macroinvertebrate shredding activity was significantly reduced in the alpha-cypermethrin treatments. In addition, the macroinvertebrate assemblage responded to the propiconazole treatments by increasing their consumption of leaf litter with lower microbial biomass, probably to compensate for the reduced nutritional quality of this leaf litter. We found no interaction between the two pesticides on macroinvertebrate shredding activity, using Independent Action as a reference model. In terms of microbial leaf decomposition rates, however, alpha-cypermethrin acted as an antagonist on propiconazole. Based on these results we emphasise the importance of considering indirect effects of pesticides in the risk assessment of surface water ecosystems.

Impact of lambda-cyhalothrin on a macroinvertebrate assemblage in outdoor experimental channels: implications for ecosystem functioning.

In this study, the impact of a single pulse of the pyrethroid lambda-cyhalothrin was tested on a macroinvertebrate assemblage consisting of Gammarus pulex, Leuctra nigra, Heptagenia sulphurea and Ancylus fluviatilis in outdoor experimental stream channels. Channels (4m long, 0.1m wide) were groundwater fed and had natural substratum. Macroinvertebrates were exposed to 10.65 or 106.5 ng L(-1) lambda cyhalothrin for 90 min in the laboratory and after 24h introduced to the experimental stream channels with four replicates of each treatment and controls. Drift samples were taken with 24-h interval for 10 days and behaviour of drifted macroinvertebrates was assessed. Microalgae biomass was measured on days 1, 5, 8 and 10 along with leaf litter decomposition using leaf packs of beech (Fagus sylvatica). Numbers of drifting G. pulex and L. nigra with reduced mobility increased significantly with concentration of lambda-cyhalothrin. Increase of algal biomass was significantly greater in stream channels with macroinvertebrates exposed to 106.5 ng L(-1) compared to controls and 10.65 ng L(-1) treatments. Accrual of microalgal biomass was significantly higher in the high concentration treatment and decomposition of leaf litter was significantly greater in control channels compared to channels with exposed macroinvertebrates. This study may apply valuable knowledge to the understanding and assessment of how pyrethroids impact ecosystem functioning in streams.