The cumulative impacts of anthropogenic stressors vary markedly along environmental gradients.

Understanding the cumulative effects of multiple stressors on biodiversity is key to managing their impacts. Stressor interactions are often studied using an additive/antagonistic/synergistic typology, aimed at identifying situations where individual stressor effects are reduced or amplified when they act in combination. Here, we analysed variation in the family richness of stream macroinvertebrates in the groups Ephemeroptera, Plecoptera and Trichoptera (EPT) at 4658 sites spanning a 32° latitudinal range in eastern Australia in relation to two largely human-induced stressors, salinity and turbidity, and two environmental gradients, temperature and slope. The cumulative and interactive effect of salinity and turbidity on EPT family richness varied across the landscape and by habitat (edge or riffle) such that we observed additive, antagonistic and synergistic outcomes depending on the environmental context. Our findings highlight the importance of understanding the consistency of multiple stressor impacts, which will involve higher-order interactions between multiple stressors and environmental factors.

Fish and macroinvertebrate assemblages reveal extensive degradation of the world's rivers.

Rivers suffer from multiple stressors acting simultaneously on their biota, but the consequences are poorly quantified at the global scale. We evaluated the biological condition of rivers globally, including the largest proportion of countries from the Global South published to date. We gathered macroinvertebrate- and fish-based assessments from 72,275 and 37,676 sites, respectively, from 64 study regions across six continents and 45 nations. Because assessments were based on differing methods, different systems were consolidated into a 3-class system: Good, Impaired, or Severely Impaired, following common guidelines. The proportion of sites in each class by study area was calculated and each region was assigned a Köppen-Geiger climate type, Human Footprint score (addressing landscape alterations), Human Development Index (HDI) score (addressing social welfare), % rivers with good ambient water quality, % protected freshwater key biodiversity areas; and % of forest area net change rate. We found that 50% of macroinvertebrate sites and 42% of fish sites were in Good condition, whereas 21% and 29% were Severely Impaired, respectively. The poorest biological conditions occurred in Arid and Equatorial climates and the best conditions occurred in Snow climates. Severely Impaired conditions were associated (Pearson correlation coefficient) with higher HDI scores, poorer physico-chemical water quality, and lower proportions of protected freshwater areas. Good biological conditions were associated with good water quality and increased forested areas. It is essential to implement statutory bioassessment programs in Asian, African, and South American countries, and continue them in Oceania, Europe, and North America. There is a need to invest in assessments based on fish, as there is less information globally and fish were strong indicators of degradation. Our study highlights a need to increase the extent and number of protected river catchments, preserve and restore natural forested areas in the catchments, treat wastewater discharges, and improve river connectivity.

Acute, diel, and annual temperature variability and the thermal biology of ectotherms.

Global warming is increasing mean temperatures and altering temperature variability at multiple temporal scales. To better understand the consequences of changes in thermal variability for ectotherms it is necessary to consider thermal variation at different time scales (i.e., acute, diel, and annual) and the responses of organisms within and across generations. Thermodynamics constrain acute responses to temperature, but within these constraints and over longer time periods, organisms have the scope to adaptively acclimate or evolve. Yet, hypotheses and predictions about responses to future warming tend not to explicitly consider the temporal scale at which temperature varies. Here, focusing on multicellular ectothermic animals, we argue that consideration of multiple processes and constraints associated with various timescales is necessary to better understand how altered thermal variability because of climate change will affect ectotherms.

Bicarbonate alone does not totally explain the toxicity from major ions of coal bed derived waters to freshwater invertebrates.

Concentrations of major ions in coal mine discharge waters and unconventional hydrocarbon produced waters derived from coal bed methane (CBM) production, are potentially harmful to freshwater ecosystems. Bicarbonate is a major constituent of produced waters from CBM and coal mining. However, little is known about the relative toxicity of differing ionic proportions, especially bicarbonate, found in these CBM waters. As all freshwater invertebrates tested are more acutely sensitive to sodium bicarbonate (NaHCO3) than sodium chloride (NaCl) or synthetic sea water, we tested the hypotheses that toxicity of CBM waters are driven by bicarbonate concentration, and waters containing a higher proportion of bicarbonate are more toxic to freshwater invertebrates than those with less bicarbonate. We compared the acute (96 h) lethal toxicity to six freshwater invertebrate species of NaHCO3 and two synthetic CBM waters, with ionic proportions representative of water from CBM wells across New South Wales (NSW) and Queensland (Qld), in Australia. The ranking of LC50 values expressed as total salinity was consistent with the hypotheses. However, when toxicity was expressed as bicarbonate concentration, the hypothesis that the toxicity of coal bed waters would be explained by bicarbonate concentration was not well supported, and other ionic components were either ameliorating or exacerbating the NaHCO3 toxicity. Our findings showed NaHCO3 was more toxic than NaCl and that the NaHCO3 proportion of synthetic CBM waters drives toxicity, however other ions are altering the toxicity of bicarbonate.

The Effects of Road De-icing Salts on Water Quality and Macroinvertebrates in Australian Alpine Areas.

The application of road de-icing salts has the potential to salinize fresh waters and degrade habitat for aquatic organisms. In the Australian Alps, the ecological effects of even small salinity increases from de-icing may be different than in North America and Europe because of (1) differences in the evolutionary history, and (2) areas with de-icing in Australia are not located in urbanized landscapes where de-icing has been largely studied elsewhere. In this study, we tried to determine the salinity increases attributable to de-icing in Australia and the effects of this increase in salinity to stream macroinvertebrates. We observed increased salt concentrations (as measured by continuous measurements of electrical conductivity (EC) and periodic measurements of chloride concentrations) in streams near two Australian ski resorts, during the snow seasons (June to September) of 2016 to 2018. The maximum EC observed in streams in salted sites near Perisher, New South Wales, was 390 µS cm-1 compared with a maximum of 26.5 µS cm-1 at unsalted sites. Lower EC values (i.e., maximum 61.1 µS cm-1) and short durations of salinity increases in streams near Falls Creek, Victoria, were not expected to cause an adverse biological response. Salt storage in the landscape was evident at salted sites near Perisher where EC was above background levels during periods of the year when no salt was applied to roads. Stream macroinvertebrate community composition differed at sites receiving run-off from road salting activities near Perisher. Abundances of Oligochaeta (worms) (up to 11-fold), Dugesiidae (flat worms) (up to fourfold), and Aphroteniinae (chironomids) (up to 14-fold) increased, whereas Leptophlebiidae (mayflies) decreased by up to 100% compared with non-salted sites. The taxa that were less abundant where de-icing salts were present tended to be the same taxa that toxicity testing revealed to be relatively salt sensitive species. This study demonstrates a causal link between de-icing salts, elevated stream salinity, and altered macroinvertebrate community composition in streams that received run-off from road de-icing activity in the Australian Alps.

Assessing the Relative Toxicity of Different Road Salts and Effect of Temperature on Salinity Toxicity: LCx Values versus No-Effect Concentration (NEC) Values.

Freshwater biota are at risk globally from increasing salinity, including increases from deicing salts in cold regions. A variety of metrics of toxicity are used when estimating the toxicity of substances and comparing the toxicity between substances. However, the implications of using different metrics are not widely appreciated. Using the mayfly Colobruscoides giganteus (Ephemeroptera: Colobruscoidea), we compare the toxicity of seven different salts where toxicity was estimated using two metrics: (1) the no-effect concentrations (NEC) and (2) the lethal concentrations for 10, 25 and 50% of the test populations (LCx). The LCx values were estimated using two different models, the classic log-logistic model and the newer toxicokinetic-toxicodynamic (TKTD) model. The NEC and both types of LCx values were estimated using Bayesian statistics. We also compared the toxicity of two salts (NaCl and CaCl2) for C. giganteus at water temperatures of 4 °C, 7 °C and 15 °C using the same metrics of toxicity. Our motivation for using a mayfly to assess salinity toxicity was because mayflies are generally salt sensitive, are ecologically important and are common in Australian (sub-)alpine streams. The temperature ranges were chosen to mimic winter, spring and summer water temperatures for Australian (sub-)alpine streams. Considering 144-h classical LCx values, we found toxicity differed between various salts, i.e., the lowest 144-h LC50 (8 mS/cm) for a salt used by a ski resort was half that of the highest 144-h LC50 from artificial marine salts and CaCl2 applied to roads (16 mS/cm). The analytical grade NaCl (as shown by 144-h LC50 value at 7 °C) was substantially more toxic (7.3 mS/cm) compared to analytical grade CaCl2 (12.5 mS/cm). Yet for NEC values, there were comparably fewer differences in toxicity between salts and none between the same salts at different temperatures. We conclude that LCx values are better suited to compare the difference in toxicity between substances or between the same substance at different test temperatures, while NEC values are better suited to estimating concentrations of substances that have no effect to the test species and endpoint measured under laboratory conditions.

Salinized rivers: degraded systems or new habitats for salt-tolerant faunas?

Anthropogenic salinization of rivers is an emerging issue of global concern, with significant adverse effects on biodiversity and ecosystem functioning. Impacts of freshwater salinization on biota are strongly mediated by evolutionary history, as this is a major factor determining species physiological salinity tolerance. Freshwater insects dominate most flowing waters, and the common lotic insect orders Ephemeroptera (mayflies), Plecoptera (stoneflies) and Trichoptera (caddisflies) are particularly salt-sensitive. Tolerances of existing taxa, rapid adaption, colonization by novel taxa (from naturally saline environments) and interactions between species will be key drivers of assemblages in saline lotic systems. Here we outline a conceptual framework predicting how communities may change in salinizing rivers. We envision that a relatively small number of taxa will be saline-tolerant and able to colonize salinized rivers (e.g. most naturally saline habitats are lentic; thus potential colonizers would need to adapt to lotic environments), leading to depauperate communities in these environments.

Pesticides reduce regional biodiversity of stream invertebrates.

The biodiversity crisis is one of the greatest challenges facing humanity, but our understanding of the drivers remains limited. Thus, after decades of studies and regulation efforts, it remains unknown whether to what degree and at what concentrations modern agricultural pesticides cause regional-scale species losses. We analyzed the effects of pesticides on the regional taxa richness of stream invertebrates in Europe (Germany and France) and Australia (southern Victoria). Pesticides caused statistically significant effects on both the species and family richness in both regions, with losses in taxa up to 42% of the recorded taxonomic pools. Furthermore, the effects in Europe were detected at concentrations that current legislation considers environmentally protective. Thus, the current ecological risk assessment of pesticides falls short of protecting biodiversity, and new approaches linking ecology and ecotoxicology are needed.

Constructing Time-Resolved Species Sensitivity Distributions Using a Hierarchical Toxico-Dynamic Model.

Classical species sensitivity distribution (SSD) is used to assess the threat to ecological communities posed by a contaminant and derive a safe concentration. It suffers from several well-documented weaknesses regarding its ecological realism and statistical soundness. Criticism includes that SSD does not take time-dependence of the data into account, that safe concentrations obtained from SSD might not be entirely protective of the target communities, and that there are issues of statistical representativity and of uncertainty propagation from the experimental data. We present a hierarchical toxico-dynamic (TD) model to simultaneously address these weaknesses: TD models incorporate time-dependence and allow improvement of the ecological relevance of safe concentrations, while the hierarchical approach affords appropriate propagation of uncertainty from the original data. We develop this model on a published data set containing the salinity tolerance over 72 h of 217 macroinvertebrate taxa, obtained through rapid toxicity testing (RTT). The shrinkage properties of the hierarchical model prove particularly adequate for modeling inhomogeneous RTT data. Taking into account the large variability in the species response, the model fits the whole data set well. Moreover, the model predicts a time-independent safe concentration below that obtained with classical SSD at 72 h, demonstrating under-protectiveness of the classical approach.

The thermal breadth of temperate and tropical freshwater insects supports the climate variability hypothesis.

Climate change involves increases in mean temperature and changes in temperature variability at multiple temporal scales but research rarely considers these temporal scales. The climate variability hypothesis (CVH) provides a conceptual framework for exploring the potential effects of annual scale thermal variability across climatic zones. The CVH predicts ectotherms in temperate regions tolerate a wider range of temperatures than those in tropical regions in response to greater annual variability in temperate regions. However, various other aspects of thermal regimes (e.g. diel variability), organisms' size and taxonomic identity are also hypothesised to influence thermal tolerance. Indeed, high temperatures in the tropics have been proposed as constraining organisms' ability to tolerate a wide range of temperatures, implying that high annual maximum temperatures would be associated with tolerating a narrow range of temperatures. We measured thermal regimes and critical thermal limits (CTmax and CTmin) of freshwater insects in the orders Ephemeroptera (mayflies), Plecoptera (stoneflies) and Trichoptera (caddisflies) along elevation gradients in streams in temperate and tropical regions of eastern Australia and tested the CVH by determining which variables were most correlated with thermal breadth (T br = CTmax - CTmin). Consistent with the CVH, T br tended to increase with increasing annual temperature range. T br also increased with body size and T br was generally wider in Plecoptera than in Ephemeroptera or Trichoptera. We also find some support for a related hypothesis, the climate extreme hypothesis (CEH), particularly for predicting upper thermal limits. We found no evidence that higher annual maximum temperature constrained individuals' abilities to tolerate a wide range of temperatures. The support for the CVH we document suggests that temperate organisms may be able to tolerate wider ranges of temperatures than tropical organisms. There is an urgent need to investigate other aspects of thermal regimes, such as diel temperature cycling and minimum temperature.

Do magnesium and chloride ameliorate high sodium bicarbonate concentrations? A comparison between laboratory and mesocosm toxicity experiments.

Increasing salinity is a concern for biodiversity in many freshwater ecosystems globally. Single species laboratory toxicity tests show major differences in freshwater organism survival depending on the specific ions that comprise salinity types and/or their ion ratios. Toxicity has been shown to be reduced by altering ionic composition, despite increasing (total) salinity. For insistence, single species tests show the toxicity of sodium bicarbonate (NaHCO3, which commonly is a large proportion of the salts from coalbeds) to freshwater invertebrates is reduced by adding magnesium (Mg2+) or chloride (Cl-). However, it is uncertain whether reductions in mortality observed in single-species laboratory tests predict effects within populations, communities and to ecosystem processes in more complex multi-species systems both natural and semi-natural. Here we report the results of an outdoor multi-species mesocosm experiment to determine if the effects of NaHCO3 are reduced by increasing the concentrations of Mg2+ or Cl- on: a) stream macroinvertebrate populations and communities; b) benthic chlorophyll-a and; c) the ecosystem process of leaf litter decomposition. We found a large effect of a high NaHCO3 concentration (≈4.45 mS/cm) with reduced abundances of multiple taxa, reduced emergence of adult insects and reduced species richness, altered community structure and increased leaf litter breakdown rates but no effect on benthic chlorophyll-a. However, despite predictions based on laboratory findings, we found no evidence that the addition of either Mg2+ or Cl- altered the effect of NaHCO3. In semi-natural environments such as mesocosms, and natural environments, organisms are subject to varying temperature and habitat factors, while also interacting with other species and trophic levels (e.g. predation, competition, facilitation), which are absent in single species laboratory tests. Thus, it should not be assumed single-species tests are good predictors of the effects of changing ionic compositions on stream biota in more natural environments.

How to characterize chemical exposure to predict ecologic effects on aquatic communities?

Reliable characterization of exposure is indispensable for ecological risk assessment of chemicals. To deal with mixtures, several approaches have been developed, but their relevance for predicting ecological effects on communities in the field has not been elucidated. In the present study, we compared nine metrics designed for estimating the total toxicity of mixtures regarding their relationship with an effect metric for stream macroinvertebrates. This was done using monitoring data of biota and organic chemicals, mainly pesticides, from five studies comprising 102 streams in several regions of Europe and South-East Australia. Mixtures of less than 10 pesticides per water sample were most common for concurrent exposure. Exposure metrics based on the 5% fraction of a species sensitivity distribution performed best, closely followed by metrics based on the most sensitive species and Daphnia magna as benchmark. Considering only the compound with the highest toxicity and ignoring mixture toxicity was sufficient to estimate toxicity in predominantly agricultural regions with pesticide exposure. The multisubstance Potentially Affected Fraction (msPAF) that combines concentration and response addition was advantageous in the study where further organic toxicants occurred. We give recommendations on exposure metric selection depending on data availability and the involved compounds.

Short-term insecticide exposure amid co-occurring stressors reduces diversity and densities in north-east Indian experimental aquatic invertebrate communities.

Globally, river pesticide concentrations are associated with regional and local stream invertebrate diversity declines. Pesticides often co-occur with elevated nutrients (e.g. nitrogen and phosphorus) and sediments related to agriculture, making their individual effects difficult to disentangle. These effects are also less well studied in Asia, than in other geographic regions. Within Asia, India is one of the largest producers and users of pesticides and has approximately 60% of total land mass used for agriculture. Here we examine the responses of Indian river invertebrate communities subjected to malathion, nutrients, and sediment additions in a semi-orthogonal design, in three sequential (through time) short-term (120 h) mesocosm experiments. Additionally, a series of single-species toxicity tests were run that used 24 h exposure and 72 h recovery to examine the sensitivity of 13 local invertebrate taxa to malathion, and 9 taxa to cypermethrin, comparing these results to those from other biogeographic regions. Mesocosm results indicate that malathion exposure had a major effect compared to other stressors on communities, with a lesser effect of nutrients and/or sediments. In mesocosms, taxa richness, total abundance and the abundance of sensitive species all declined associated with malathion concentrations. Comparisons of organism sensitivities from other geographic locations and those in the current paper suggest taxa in India are relatively tolerant to malathion and cypermethrin. Our results further reinforce that the high observed aquatic pesticide concentrations known to occur in Asian freshwater ecosystems are likely to be negatively affecting biodiversity, homogenising biota towards those most stress tolerant.

Thresholds for the effects of pesticides on invertebrate communities and leaf breakdown in stream ecosystems.

We compiled data from eight field studies conducted between 1998 and 2010 in Europe, Siberia, and Australia to derive thresholds for the effects of pesticides on macroinvertebrate communities and the ecosystem function leaf breakdown. Dose-response models for the relationship of pesticide toxicity with the abundance of sensitive macroinvertebrate taxa showed significant differences to reference sites at 1/1000 to 1/10,000 of the median acute effect concentration (EC50) for Daphnia magna, depending on the model specification and whether forested upstream sections were present. Hence, the analysis revealed effects well below the threshold of 1/100 of the EC50 for D. magna incorporated in the European Union Uniform Principles (UP) for registration of single pesticides. Moreover, the abundances of sensitive macroinvertebrates in the communities were reduced by 27% to 61% at concentrations related to 1/100 of the EC50 for D. magna. The invertebrate leaf breakdown rate was positively linearly related to the abundance of pesticide-sensitive macroinvertebrate species in the communities, though only for two of the three countries examined. We argue that the low effect thresholds observed were not mainly because of an underestimation of field exposure or confounding factors. From the results gathered we derive that the UP threshold for single pesticides based on D. magna is not protective for field communities subject to multiple stressors, pesticide mixtures, and repeated exposures and that risk mitigation measures, such as forested landscape patches, can alleviate effects of pesticides.

Leaf litter breakdown along an elevational gradient in Australian alpine streams.

The breakdown of allochthonous organic matter, is a central step in nutrient cycling in stream ecosystems. There is concern that increased temperatures from climate change will alter the breakdown rate of organic matter, with important consequences for the ecosystem functioning of alpine streams. This study investigated the rate of leaf litter breakdown and how temperature and other factors such as microbial and invertebrate activities influenced this over elevational and temporal gradients. Dried leaves of Snow Gum (Eucalyptus pauciflora) and cotton strips were deployed in coarse (6 mm), and fine (50 µm) mesh size bags along an 820 m elevation gradient. Loss of mass in leaf litter and cotton tensile strength per day (k per day), fungal biomass measured as ergosterol concentration, invertebrate colonization of leaf litter, and benthic organic matter (mass and composition) were determined. Both microbial and macroinvertebrate activities were equally important in leaf litter breakdown with the abundance of shredder invertebrate taxa. The overall leaf litter breakdown rate and loss of tensile strength in cotton strips (both k per day) were greater during warmer deployment periods and at lower elevations, with significant positive relationships between mean water temperature and leaf breakdown and loss of tensile strength rate, but no differences between sites, after accounting for the effects of temperature. Despite considerably lower amounts of benthic organic matter in streams above the tree line relative to those below, shredders were present in coarse mesh bags at all sites. Ergosterol concentration was greater on leaves in coarse mesh bags than in fine mesh bags, implying differences in the microbial communities. The importance of water temperatures on the rate of leaf litter breakdown suggests the potential effects of climate change-induced temperature increases on ecological processes in such streams.

Effects of pesticides monitored with three sampling methods in 24 sites on macroinvertebrates and microorganisms.

Grab water samples, sediment samples, and 2,2,4-trimethylpentane passive samplers (TRIMPS) were used to determine the exposure to 97 pesticides in 24 southeast Australian stream sites over 5 months. Macroinvertebrate communities and selected microorganisms (bacteria, flagellates, ciliates, amoebas, nematodes, and gastrotrichs) were sampled to detect relationships with pesticide toxicity. Sediment samples had the highest estimated toxicities in terms of toxic units (TU) for Daphnia magna (TUDM) and for Selenastrum capricornutum (TUSC). The pesticide-selective SPEARpesticides and the general SIGNAL index for macroinvertebrates exhibited negative linear relationships (r(2) = 0.67 and 0.36, respectively) with pesticide contamination in terms of log maximum TUDM (log mTUDM), suggesting macroinvertebrate community change due to pesticide exposure. Pesticide contamination was the only measured variable explaining variation in ecological quality. Variation in the densities of several microbial groups was best explained by environmental variables other than log TUs. The log mTUDM values derived from sediment concentrations were most important to establish a link with effects on macroinvertebrates, whereas log mTUDM of grab water samples had only minor contribution. Current-use insecticides and fungicides can affect macroinvertebrate communities and monitoring of sediment and continuous water sampling is needed to detect these effects.

Sensitivity and specificity of macroinvertebrate responses to gradients of multiple agricultural stressors.

Environmental degradation of rivers in agricultural landscapes is typically caused by multiple co-occurring stressors, but how interactions among stressors affect freshwater ecosystems is poorly understood. Therefore, we investigated the sensitivity and specificity of several measures of benthic macroinvertebrate community response to the individual and combined effects of the pesticide sulfoxaflor (SFX), increased sand sedimentation and elevated nutrients using outdoor recirculating mesocosms. Among the single stressor treatments, nutrients had no observable impact and sand only affected one community response measure compared to controls. High SFX levels had the largest effects on benthic macroinvertebrate communities, negatively affecting six of seven macroinvertebrate response measures. Sulfoxaflor had similar adverse effects on biota when in combination with sand and nutrients in the multi-stressor treatment, suggesting that generally SFX has overwhelming and pervasive effects irrespective of the presence of the other stressors. In contrast to SFX, elevated nutrients had no detectable effect on macroinvertebrate communities, likely as a consequence of nutrients being rapidly taken up by bacteria rather than by benthic algae. Elevated sand sedimentation increased the negative effects of SFX on sediment sensitive taxa, but generally had limited biological effects. This was despite the levels of sedimentation in our treatments being at concentrations that have caused large impacts in other studies. This research points to direct and rapid toxic effects of SFX on stream macroinvertebrates, contrasting with effects of the other stressors. This study emphasises that pesticide effects could be misattributed to other common freshwater stressors, potentially focussing restoration actions on a stressor of lesser importance.

Can SPEcies At Risk of pesticides (SPEAR) indices detect effects of target stressors among multiple interacting stressors?

Pesticides are increasingly recognised as a threat to freshwater biodiversity, but their specific ecological effects remain difficult to distinguish from those of co-occurring stressors and environmental gradients. Using mesocosms we examined the effects of an organophosphate insecticide (malathion) on stream macroinvertebrate communities concurrently exposed to a suite of stressors typical of streams in agricultural catchments. We assessed the specificity of the SPEcies At Risk index designed to determine pesticide effects in mesocosm trials (SPEARmesocosm). This index determines the log abundance proportion of taxa that are considered physiologically sensitive to pesticides. Geographic variation in pesticide sensitivity within taxa, coupled with variation between pesticides and the effects of co-occurring stressors may decrease the accuracy of SPEARmesocosm. To examine this, we used local pesticide sensitivity assessments based on rapid toxicity tests to develop two new SPEAR versions to compare to the original SPEARmesocosms index using mesocosm results. We further compared these results to multivariate analyses and community indices (e.g. richness, abundance, Simpson's diversity) commonly used to assess stressor effects on biota. To assess the implications of misclassifying species sensitivity on SPEAR indices we used a series of simulations using artificial data. The impacts of malathion were detectable using SPEARmesocosm, and one of two new SPEAR indices. All three of the SPEAR indices also increased when exposed to other agricultural non-pesticide stressors, and this change increased with greater pesticide concentrations. Our results support that interactions between other non-pesticide stressors with pesticides can affect SPEAR performance. Multivariate analysis and the other indices used here identified a significant effect of malathion especially at high concentrations, with little or no evidence of effects from the other agricultural stressors.