Combined Effects Of Stressors Research Articles

Human rights in countries of origin and the mental health of migrants to Canada.

This study explores the effect of human rights violations in countries of origin on migrants' mental health, using archival data on human rights violations from 1970-2011, merged to a representative probability sample of 2412 adults living in a large Canadian metropolitan area. The context of exit is defined at the country level, as opposed to self-reported individual experiences of trauma. While most studies start from a question about direct exposure to human rights violations, they may miss the effect of the national-level social context - threat, instability, disruption of lives, and uncertainty - on mental health. Findings indicate that high levels of human rights violations in countries of origin have long-term effects on migrants’ mental health. The impact of human rights violations is substantially explained by the combined effect of stressors both before and after migration, suggesting a cumulative process of stress proliferation following this context of exit.

Effects of combined stressors to cadmium and high temperature on antioxidant defense, apoptotic cell death, and DNA methylation in zebrafish (Danio rerio) embryos.

Fish are frequently affected by environmental stressors, such as temperature changes and heavy metal exposure, in aquatic ecosystems. In this study, we evaluated the combined effects of cadmium (Cd) toxicity and temperature (rearing temperature of 26°C and heat stress at 34°C) on zebrafish (Danio rerio) embryos. The survival and heart rates of zebrafish embryos decreased at relatively high Cd concentrations of 0.07 and 0.1mgL-1. Abnormal morphology was induced by exposure to a combination of Cd toxicity and heat stress. The yolk sac edema size was not significantly different between the control- and Cd-treated groups. Cd exposure induced reactive oxygen species (ROS) production and cell death in the live zebrafish. High temperature (34°C) triggered Cd-induced cell death and intracellular ROS production to a greater extent than the rearing temperature of 26°C. Transcriptional levels of six genes-CAT, SOD, p53, BAX, Dnmt1, and Dnmt3b-were investigated. The mRNA expression of CAT and SOD, molecular indicators of oxidative stress, was increased significantly at 34°C after Cd exposure. The mRNA expression of CAT was more sensitive to temperature than that of SOD in Cd-treated zebrafish. p53 and BAX, apoptosis-related genes, were upregulated upon combined exposure to high temperature and Cd. In addition, at 34°C, the expression of Dnmt1 and Dnmt3b transcripts, markers of DNA methylation, was increased upon exposure of zebrafish to all concentrations of Cd. Overall, these results suggest that high temperature facilitates the potential role of Cd toxicity in the transcriptional regulation of genes involved in the antioxidant system, apoptosis, and DNA methylation.

Population context matters: Predicting the effects of metabolic stress mediated by food availability and predation with an agent- and energy budget-based model

Population effects of stressors, such as toxic chemicals or increased temperatures, affecting the energy budgets of organisms are mediated by predation pressure and food availability. However, these two population contexts have mostly been considered separately. Moreover, because the sensitivity of the different pathways of energy to stress may differ, it is difficult to predict combined stressor effects. We used an agent-based model of fathead minnows (Pimephales promelas) to infer the population-level impacts of a hypothetical, sublethal stressor that affects an individual's metabolism (growth, reproduction, maintenance, or assimilation) in systems in which population size is controlled by different combinations of food availability and predation. We found that population-level effects are rarely directly proportional to individual-level effects, and were greater when the stressor impacted assimilation and populations were predation-controlled. Our results suggest that individual-level measurements alone are insufficient for inferring population-level impacts of stressors and that accurate inference hinges on insight into how populations are regulated. We suggest incorporating individual-level data into mechanistic models that take into account both the energy budgets of individuals and the population-level context.

Multiple stressor effects on coral reef ecosystems

Global climate change has profound implications on species distributions and ecosystem functioning. In the coastal zone, ecological responses may be driven by various biogeochemical and physical environmental factors. Synergistic interactions can occur when the combined effects of stressors exceed their individual effects. The Red Sea, characterized by strong gradients in temperature, salinity, and nutrients along the latitudinal axis provides a unique opportunity to study ecological responses over a range of these environmental variables. Using multiple linear regression models integratingin situ, satellite and oceanographic data, we investigated the response of coral reef taxa to local stressors and recent climate variability. Taxa and functional groups responded to a combination of climate (temperature, salinity, air-sea heat fluxes, irradiance, wind speed), fishing pressure and biogeochemical (chlorophyll a and nutrients - phosphate, nitrate, nitrite) factors. The regression model for each species showed interactive effects of climate, fishing pressure and nutrient variables. The nature of the effects (antagonistic or synergistic) was dependent on the species and stressor pair. Variables consistently associated with the highest number of synergistic interactions included heat flux terms, temperature, and wind speed followed by fishing pressure. Hard corals and coralline algae abundance were sensitive to changing environmental conditions where synergistic interactions decreased their percentage cover. These synergistic interactions suggest that the negative effects of fishing pressure and eutrophication may exacerbate the impact of climate change on corals. A high number of interactions were also recorded for algae, however for this group, synergistic interactions increased algal abundance. This study is unique in applying regression analysis to multiple environmental variables simultaneously to understand stressor interactions in the field. The observed responses have important implications for understanding climate change impacts on marine ecosystems and whether managing local stressors, such as nutrient enrichment and fishing activities, may help mitigate global drivers of change.

Interacting with change: Diet mediates how larvae respond to their thermal environment

Abstract Temperature and nutrition are amongst the most common environmental challenges faced by organisms and will become increasingly so with ongoing climate change. While we have learnt a great deal about how temperature and nutrition affect life‐history traits on their own, we know very little about their combined effect on animal performance. Given that animals in the wild are likely to experience changes in both their thermal and nutritional conditions, we need to understand how interactions between these conditions shape an animal's response if we hope to mitigate the effects of environmental change. In the present research, we investigated the combined effects of nutrition and temperature on key life‐history traits in Drosophila melanogaster. Using nutritional geometry, developing larvae were exposed to a range of diets varying in their protein and carbohydrate content and to one of two developmental temperature regimes (25°C and 28°C). We then examined key life‐history traits: development time, viability, and two estimates of body size—wing and femur size. We found that developmental temperature significantly changed the response to nutrition for all traits. Increased temperature led to more restricted trait optima for all traits and exacerbated the negative effects of carbohydrate‐rich diets, resulting in harsher trade‐offs between life‐history traits. For example, at 25°C there were more diets that led to high viability, fast development and large body size than at 28°C. However, for the diets that produced the best outcomes for each trait, temperature had less of an effect. These findings highlight the importance of studying the effects of combined stressors when assessing animals' responses to changing environmental conditions. A free Plain Language Summary can be found within the Supporting Information of this article.

Effects of sublethal doses of clothianidin and/or V. destructor on honey bee (Apis mellifera) self-grooming behavior and associated gene expression

Little is known about the combined effects of stressors on social immunity of honey bees (Apis mellifera) and related gene expression. The interaction between sublethal doses of a neurotoxin, clothianidin, and the ectoparasite, Varroa destructor, was examined by measuring differentially expressed genes (DEGs) in brains, deformed wing virus (DWV) and the proportion and intensity of self-grooming. Evidence for an interaction was observed between the stressors in a reduction in the proportion of intense groomers. Only the lowest dose of clothianidin alone reduced the proportion of self-groomers and increased DWV levels. V. destructor shared a higher proportion of DEGs with the combined stressors compared to clothianidin, indicating that the effects of V. destructor were more pervasive than those of clothianidin when they were combined. The number of up-regulated DEGs were reduced with the combined stressors compared to clothianidin alone, suggesting an interference with the impacts of clothianidin. Clothianidin and V. destructor affected DEGs from different biological pathways but shared impacts on pathways related to neurodegenerative disorders, like Alzheimer’s, which could be related to neurological dysfunction and may explain their negative impacts on grooming. This study shows that the combination of clothianidin and V. destructor resulted in a complex and non-additive interaction.

Cumulative effects of municipal effluent and parasite infection in yellow perch: A field study using high-throughput RNA-sequencing

Multiple metabolic, immune and reproductive effects have been reported in fish residing in effluent-impacted sites. Natural stressors such as parasites also have been shown to impact the responses of organisms to chronic exposure to municipal effluent in the St. Lawrence River (Quebec, Canada). In order to comprehensively evaluate the cumulative impacts of anthropogenic and natural stressors on the health of yellow perch, differential mRNA transcription profiles were examined in juvenile females collected from effluent-impacted and upstream sites with low or high infection levels of the larval trematode Apophallus brevis. Transcriptomics was used to identify biological pathways associated with environmental exposure. In total, 3463 isoforms were differentially transcribed between sites. Patterns reflecting the combined effects of stressors were numerically dominant, with a majority of downregulated transcripts (68%). The differentially expressed transcripts were associated with 27 molecular and cellular functions ranging from cellular development to xenobiotic metabolism and were involved in the development and function of 13 organ systems including hematological, hepatic, nervous, reproductive and endocrine systems. Based on RNA-seq results, sixteen genes were measured by qPCR. Significant differences were observed for six genes in fish exposed to both stressors combined, whereas parasites and effluent individually impacted the transcription of one gene. Lysozyme activity, lipid peroxidation, retinol-binding protein and glucose-6-phosphate dehydrogenase were selected as potential biomarkers of effects to study specific pathways of interest. Lipid peroxidation in perch liver was different between sites, parasite loads, and for combined stressors. Overall, results indicated that juvenile yellow perch responded strongly to combined parasite and effluent exposure, suggesting cumulative effects on immune responses, inflammation and lipid metabolism mediated by retinoid receptors. The present study highlight the importance of using a comprehensive approach combining transcriptomics and endpoints measured at higher levels of biological organization to better understand cumulative risks of contaminants and pathogens in aquatic ecosystems.

Combined effects of cadmium exposure and temperature on the annual killifish (Nothobranchius furzeri).

Freshwater organisms are increasingly exposed to combinations of stressors. However, because it is time-consuming and costly, research on the interaction of stressors, such as compound toxicity and global warming on vertebrates, is scarce. Studies on multigenerational effects of these combined stressors are almost nonexistent. In the present study, we tested the combined effects of 4 °C warming and cadmium (Cd) exposure on life-history traits, biomarkers, bioaccumulation, and multigenerational tolerance in the turquoise killifish, Nothobranchius furzeri. The extremely short life cycle of this vertebrate model allows for assessment of sublethal and multigenerational effects within 4 mo. The applied Cd concentrations had only limited effects on the measured endpoints, which suggests that N. furzeri is more resistant to Cd than fathead minnow and rainbow trout. In contrast, the temperature increase of 4 °C was stressful: it delayed female maturation and lowered adult mass and fecundity. Finally, indications of synergistic effects were found on peak fecundity and embryonic survival. Overall, these results indicate the importance of studying chronic and multigenerational effects of combined stressors. Environ Toxicol Chem 2018;37:2361-2371. © 2018 SETAC.

Interactions between perceived exertion and thermal perception in the heat in endurance athletes

IntroductionThe study aimed to investigate how a distortion of perceived exertion in the heat may affect, during a self-paced cycling exercise preceded by prior cognitive task, the thermal perception and the subsequent regulation of power output in high level athletes. MethodsEleven endurance trained male athletes completed four experimental sessions including a 30-min fixed-RPE (15-Hard) cycling exercise in neutral (TMP-22 °C) and hot (HOT-37 °C) conditions, following a 60-min incongruent Stroop task (EXP) or passively watching documentary films (CON). Central and peripheral performances of the knee extensors were assessed before the cognitive task and after the exercise. ResultsAlthough mental demand and effort were higher in EXP (P < 0.05), no effect of prior cognitive task was observed on subjective feelings of mental fatigue or decline in power output at a fixed RPE. Average exercise intensity was lower in HOT than TMP (3.14 ± 0.09 W⋅kg−1vs. 3.42 ± 0.10 W⋅kg−1 respectively, P < 0.05). Skin temperature and warmth sensations were higher in HOT throughout the exercise (P < 0.05) but not thermal comfort. Central and peripheral parameters were not affected more in HOT than in TMP. ConclusionAlthough the effects of combined stressors on the distortion of perceived exertion could not be verified, the greater decline in power output recorded in HOT than TMP suggest a high contribution of both perceptual and cardiovascular responses in the regulation of work rate when the subject is in mild hyperthermia.

Simultaneous exposure to a pulsed and a prolonged anthropogenic stressor can alter consumer multifunctionality

Ecosystems face multiple anthropogenic threats globally, and the effects of these environmental stressors range from individual‐level organismal responses to altered system functioning. Understanding the combined effects of stressors on process rates mediated by individuals in ecosystems would greatly improve our ability to predict organismal multifunctionality (e.g. multiple consumer‐mediated functions). We conducted a laboratory experiment to test direct and indirect, as well as immediate and delayed effects of a heat wave (pulsed stress) and micropollutants (MPs) (prolonged stress) on individual consumers (the great pond snail Lymnaea stagnalis) and their multifunctionality (i.e. consumption of basal resources, growth, reproduction, nutrient excretion and organic‐matter cycling). We found that stressful conditions increased the process rates of multiple functions mediated by individual consumers. Specifically, the artificial heat wave increased process rates in the majority of the quantified functions (either directly or indirectly), whereas exposure to MPs increased consumption of basal resources which led to increases in the release of nutrients and fine particulate organic matter. Moreover, snails exposed to a heat wave showed decreased reproduction and nutrient excretion after the heat‐wave, indicating the potential for ecologically relevant delayed effects. Our study indicates that the immediate and delayed effects of stressors on individual organisms may directly and indirectly impact multiple ecosystem functions. In particular, delayed effects of environmental stress on individual consumers may cumulatively impede recovery due to decreased functioning following a perturbation. Reconciling these results with studies incorporating responses at higher levels of biological complexity will enhance our ability to forecast how individual responses upscale to ecosystem multifunctionality.

Protocol for Acute and Chronic Ecotoxicity Testing of the Turquoise Killifish Nothobranchius furzeri.

The killifish Nothobranchius furzeri is an emerging model organism in the field of ecotoxicology and its applicability in acute and chronic ecotoxicity testing has been demonstrated. Overall, the sensitivity of the species to toxic compounds is in the range with, or higher than, that of other model species. This work describes protocols for acute, chronic, and multigenerational bioassays of single and combined stressor effects on N. furzeri. Due to its short maturation time and life-cycle, this vertebrate model allows the study of endpoints such as maturation time and fecundity within four months. Transgenerational full life-cycle exposure trials can be performed in as little as 8 months. Since this species produces eggs that are drought-resistant and remain viable for years, the on-site culture of the species is not needed but individuals can be recruited when required. The protocols are designed to measure life-history traits (mortality, growth, fecundity, weight) and critical thermal maximum.

Informing recovery in a human-transformed landscape: Drought-mediated coexistence alters population trends of an imperiled salamander and invasive predators

Understanding the additive or interactive threats of habitat transformation and invasive species is critical for conservation, especially where climate change is expected to increase the severity or frequency of drought. In the arid southwestern USA, this combination of stressors has caused widespread declines of native aquatic and semi-aquatic species. Achieving resilience to drought and other effects of climate change may depend upon continued management, so understanding the combined effects of stressors is important. We used Bayesian hierarchical models fitted with 10-years of pond-based monitoring surveys for the federally-endangered Sonoran Tiger Salamander (Ambystoma mavortium stebbinsi) and invasive predators (fishes and American Bullfrogs, Lithobates catesbeianus) that threaten native species. We estimated trends in occupancy of salamanders and invasive predators while accounting for hydrological dynamics of ponds, then used a two-species interaction model to directly estimate how invasive predators affected salamander occupancy. We also tested a conceptual model that predicted that drought, by limiting the distribution of invasive predators, could ultimately benefit native species. Even though occupancy of invasive predators was stationary and their presence in a pond reduced the probability of salamander presence by 23%, occupancy of Sonoran Tiger Salamanders increased, annually, by 2.2%. Occupancy of salamanders and invasive predators both declined dramatically following the 5th consecutive year of drought. Salamander occupancy recovered quickly after return to non-drought conditions, while occupancy of invasive predators remained suppressed. Models that incorporated three time-lagged periods (1 to 4years) of local moisture conditions confirmed that salamanders and invasive predators responded differently to drought, reflecting how life-history strategies shape responses to disturbances. The positive 10-year trend in salamander occupancy and their rapid recovery after drought provided partial support for the hypothesis of drought-mediated coexistence with invasive predators. These results also suggest management opportunities for conservation of the Sonoran Tiger Salamander and other imperiled organisms in human-transformed landscapes.

Combined effects of elevated pCO2, temperature, and starvation stress on larvae of a large tropical marine fish

Ocean acidification and other environmental changes pose an ecological challenge to marine organisms globally. Although the youngest life stages of these organism are likely to be most affected, a limited number of studies of larval fishes have investigated the effects of combined stressors. We conducted two experiments on larval cobia (Rachycentron canadum) raised under combinations of elevated pCO2 and increased temperature or starvation stress. Larvae responded to individual CO2, temperature, and rationing treatments, and there was a negative effect of elevated pCO2 on starvation resistance, but few synergistic effects of combined stressors. Elevated pCO2 (1700–2100 μatm pCO2) caused a transient but significant reduction in larval standard length (SL), growth rate, and development rate, while warmer temperature (32 vs. 27 °C) caused a consistent increase in SL, development rate, and swimming ability. Larval condition (RNA:DNA ratio) was unaffected by elevated pCO2 although larvae fed a 25% ration had significantly reduced SL, growth rate, and development rate. Under complete feeding cessation, larvae in elevated-pCO2 seawater demonstrated lower starvation resistance, indicating that acidification may increase starvation risk in a patchy marine environment. Overall, our results indicate that larval cobia are resistant to any major direct impact of combined elevated pCO2 and temperature or rationing stress.

Analysing the impact of multiple stressors in aquatic biomonitoring data: A ‘cookbook’ with applications in R

Multiple stressors threaten biodiversity and ecosystem integrity, imposing new challenges to ecosystem management and restoration. Ecosystem managers are required to address and mitigate the impact of multiple stressors, yet the knowledge required to disentangle multiple-stressor effects is still incomplete. Experimental studies have advanced the understanding of single and combined stressor effects, but there is a lack of a robust analytical framework, to address the impact of multiple stressors based on monitoring data. Since 2000, the monitoring of Europe's waters has resulted in a vast amount of biological and environmental (stressor) data of about 120,000 water bodies. For many reasons, this data is rarely exploited in the multiple-stressor context, probably because of its rather heterogeneous nature: stressors vary and are mixed with broad-scale proxies of environmental stress (e.g. land cover), missing values and zero-inflated data limit the application of statistical methods and biological indicators are often aggregated (e.g. taxon richness) and do not respond stressor-specific. Here, we present a ‘cookbook’ to analyse the biological response to multiple stressors using data from biomonitoring schemes. Our ‘cookbook’ includes guidance for the analytical process and the interpretation of results. The ‘cookbook’ is accompanied by scripts, which allow the user to run a stepwise analysis based on his/her own data in R, an open-source language and environment for statistical computing and graphics. Using simulated and real data, we show that the recommended procedure is capable of identifying stressor hierarchy (importance) and interaction in large datasets. We recommend a minimum number of 150 independent observations and a minimum stressor gradient length of 75% (of the most relevant stressor's gradient in nature), to be able to reliably rank the stressor's importance, detect relevant interactions and estimate their standardised effect size. We conclude with a brief discussion of the advantages and limitations of this protocol.

Synergistic and additive effects of drought stress and simulated herbivory on two goldenrods, Solidago altissima and S. gigantea

Understanding the combined effects of stressors on plants is important for understanding how they will tolerate herbivory and other damage under unfavorable conditions. When two stresses have synergistic effects, plants may experience particularly strong impacts. We examined individual and combined effects of drought stress and clipping (simulated herbivory) on two species of goldenrods (Solidago altissima L. and S. gigantea Ait.). Each stress reduced production of most plant structures, with drought stress having stronger impacts than clipping. The effects of the two stresses were additive for S. gigantea but synergistic for S. altissima, at least for aboveground biomass and inflorescence biomass. Both species, when under stress, reallocated resources toward asexual reproduction (rhizomes) and away from sexual reproduction (inflorescences). Our results suggest that even closely related plants may tolerate damage differently when under abiotic stresses, and that predicting the additive vs. synergistic action of combined stresses will be difficult.

Induced community tolerance of periphyton towards combined salt and toxic stress

Summary Aquatic ecosystems are exposed to multiple stressors requiring better integration of distinct disciplines such as ecotoxicology and ecology. Stressors select for increasing community tolerance which can be evidenced by the concept of ‘pollution‐induced community tolerance’ (PICT). However, for analysing combined stressor effects on community tolerance, suitable metabolic tests are needed. Salinisation and toxic exposure often co‐occur in aquatic systems. We aimed to develop a metabolic test to quantify salt tolerance by using proline as an indicator. This method was applied in microcosms, exposing periphyton to salt and an herbicide, individually and in combination. Proline was a suitable marker for salt stress in periphyton. Long‐term exposure to single stressors resulted in toxicant‐ and salt‐induced community tolerance, but not in co‐tolerances between the stressors. Induction ratios of proline suggest that proline may have different protective functions in organisms depending on stressor quality and resulting in contrasting tolerance patterns in periphyton. Biochemical markers provide a set of metabolic tests needed for a broader application of the PICT approach in ecology and ecotoxicology, especially for analysing combined stress. Induced community tolerance may therewith improve ecological relevance in environmental risk assessment of toxicants.

Epigenome: A Biomarker or Screening Tool to Evaluate Health Impact of Cumulative Exposure to Chemical and Non-Chemical Stressors

Current risk assessment practices and toxicity information are hard to utilize for assessing the health impact of combined or cumulative exposure to multiple chemical and non-chemical stressors encountered in the “real world” environment. Non-chemical stressors such as heat, radiation, noise, humidity, bacterial and viral agents, and social factors, like stress related to violence and socioeconomic position generally cannot be currently incorporated into the risk assessment paradigm. The Science and Decisions report released by the National Research Council (NRC) in 2009 emphasized the need to characterize the effects of multiple stressors, both chemical and non-chemical exposures. One impediment to developing information relating such non-chemical stressors to health effects and incorporating them into cumulative assessment has been the lack of analytical tools to easily and quantitatively monitor the cumulative exposure to combined effects of stressors over the life course.

Multiple‐stressor effects on stream invertebrates: a mesocosm experiment manipulating nutrients, fine sediment and flow velocity

Summary Land‐use changes have degraded ecosystems worldwide. A particular concern for freshwater biodiversity and ecosystem function are stressors introduced by intensified agriculture. Typically several stressors affect freshwater ecosystems simultaneously. However, the combined effects of these multiple stressors on streams and rivers are still poorly understood, yet of critical importance to improve freshwater management. We investigated responses of benthic macroinvertebrates to three globally important agricultural stressors affecting streams (nutrient enrichment, fine sediment deposition and reduced current velocity), using 64 stream mesocosms (full‐factorial 2 × 2 × 2 design, eight replicates of each treatment combination) established on the banks of the Breitenbach Stream (Hesse, Germany). The experiment ran for 1 month (16 days of colonisation, 14 days of manipulations), and all invertebrates in the mesocosms were collected at the end of this period. Fourteen of the 17 studied invertebrate response variables were affected by one or more stressors each. Negative effects on richness or abundance of pollution‐sensitive Ephemeroptera, Plecoptera and Trichoptera (EPT) taxa were particularly common. Overall, both sediment addition and stream flow velocity reduction had pervasive and strong effects. Responses to sediment addition were mostly negative, whereas decreased current velocity reduced several EPT metrics but increased the abundances of some of the other common taxa. Nutrient enrichment had few effects, but these were consistently negative. Combined stressor effects were mainly additive, with only two interactions found in total, both between reduced velocity and nutrients (on the crustacean Gammarus spp. and ceratopogonid midges). This finding implies that multiple‐stressor responses may be predicted from knowledge of single‐stressor effects in this stream community (unlike the often synergistic or antagonistic responses observed elsewhere). However, further taxon‐specific responses and interactions among stressors may have been obscured by limited taxonomic resolution, especially for the numerically dominant Chironomidae. Genetic approaches are required to address this limitation in the future.

Macroalgal responses to ocean acidification depend on nutrient and light levels

Ocean acidification may benefit algae that are able to capitalise on increased carbon availability for photosynthesis but is expected to have adverse effects on calcified algae through dissolution. Shifts in dominance between primary producers will have knock-on effects on marine ecosystems and will likely vary regionally, depending on factors such as irradiance (light vs shade) and nutrient levels (oligotrophic vs eutrophic). Thus experiments are needed to evaluate interactive effects of combined stressors in the field. In this study, we investigated the physiological responses of macroalgae near a CO2 seep in oligotrophic waters off Vulcano (Italy). The algae were incubated in situ at 0.2 m depth using a combination of three mean CO2 levels (500, 700-800 and 1200 µatm CO2), two light levels (100 and 70% of surface irradiance) and two nutrient levels of N, P, and K (enriched vs non-enriched treatments) in the non-calcified macroalga Cystoseira compressa (Phaeophyceae, Fucales) and calcified Padina pavonica (Phaeophyceae, Dictyotales). A suite of biochemical assays and in vivo chlorophyll a fluorescence parameters showed that elevated CO2 levels benefitted both of these algae, although their responses varied depending on light and nutrient availability. In C. compressa, elevated CO2 treatments resulted in higher carbon content and antioxidant activity in shaded conditions both with and without nutrient enrichment - they had more Chla, phenolic and fucoxanthin with nutrient enrichment and higher quantum yield (Fv/Fm) and photosynthetic efficiency (αETR) without nutrient enrichment. In P. pavonica, elevated CO2 treatments had higher carbon content, Fv/Fm, αETR, and Chla regardless of nutrient levels - they had higher concentrations of phenolic compounds in nutrient enriched, fully-lit conditions and more antioxidants in shaded, nutrient enriched conditions. Nitrogen content increased significantly in fertilised treatments, confirming that these algae were nutrient

Toxic effects of combined stressors on Daphnia pulex: Interactions between diazinon, 4-nonylphenol, and wastewater effluent.

Contaminant exposure in aqueous systems typically involves complex chemical mixtures. Given the large number of compounds present in the environment, it is critical to identify hazardous chemical interactions rapidly. The present study utilized a prototype for a novel high-throughput assay to quantify behavioral changes over time to identify chemical interactions that affect toxicity. The independent and combined effects of 2 chemicals, diazinon (an insecticide) and 4-nonylphenol (a detergent metabolite), on the swimming behavior of the freshwater crustacean Daphnia pulex were examined. Cumulative distance and change in direction were measured repeatedly via optical tracking over 90 min. Exposure to low concentrations of diazinon (0.125-2 µM) or 4-nonylphenol (0.25-4 µM) elicited significant concentration- and time-dependent effects on swimming behavior. Exposure to 0.5 µM 4-nonylphenol alone did not significantly alter mean cumulative distance but did elicit a small, significant increase in mean angle, the measure of change in direction. When 0.5 µM 4-nonylphenol was used in combination with diazinon (0.125-0.5 µM), it augmented the adverse impact of diazinon on the swimming behavior of Daphnia. Additionally, enhanced sensitivity to diazinon was observed in animals exposed to treated wastewater effluent for 24 h prior to a diazinon challenge. The present experiments demonstrate that exposure to 4-nonylphenol and complex chemical mixtures (e.g., treated wastewater) can enhance the toxicity of exposure to the insecticide diazinon.