Seasonal changes in diet, immune function, and oxidative stress in three passerines inhabiting a Mediterranean climate.

Oxidative status and immune function are energy-demanding traits closely linked to diet composition, particularly resource availability and nutritional value. In seasonal environments, nutrient availability and diet quality fluctuate, potentially influencing these traits. However, limited evidence exists regarding these dietary effects on immune function in seasonal environments. In this study, we employed stable isotope analysis to assess the impact of seasonal changes in niche width and trophic level (i.e., δ15N) on two immune variables (hemolysis and hemagglutination scores) and two oxidative status parameters (lipid peroxidation and total antioxidant capacity) in three passerine species: Zonotrichia capensis (omnivorous), Troglodytes aedon (insectivorous), and Spinus barbatus (granivorous). We found that hemolysis scores varied seasonally in Z. capensis, with higher values in winter compared to summer. Total antioxidant capacity (TAC) also increased during the winter in Z. capensis and S. barbatus. The isotopic niche width for Z. capensis and S. barbatus was smaller in winter than in summer, with the omnivorous species exhibiting a decrease in δ15N. Despite the seasonal shifts in ecological and physiological traits in Z. capensis, we identified no correlation between immune response and TAC with trophic level. In contrast, in the granivorous S. barbatus, the lower trophic level resulted in an increase in TAC without affecting immunity. Our findings revealed that dietary shifts do not uniformly impact oxidative status and immune function across bird species, highlighting species-specific responses to seasonal changes. This underscores the importance of integrating ecological and evolutionary perspectives when examining how diet shapes avian immunity and oxidative balance.

Modeling heterothermic fitness landscapes in a marsupial hibernator using changes in body composition.

Hibernation is an adaptive strategy that allows animals to enter a hypometabolic state, conserving energy and enhancing their fitness by surviving harsh environmental conditions. However, addressing the adaptive value of hibernation, at the individual level and in natural populations, has been challenging. Here, we applied a non-invasive technique, body composition analysis by quantitative magnetic resonance (qMR), to calculate energy savings by hibernation in a population of hibernating marsupials (Dromiciops gliroides). Using outdoor enclosures installed in a temperate rainforest, and measuring qMR periodically, we determined the amount of fat and lean mass consumed during a whole hibernation cycle. With this information, we estimated the daily energy expenditure of hibernation (DEEH) at the individual level and related to previous fat accumulation. Using model selection approaches and phenotypic selection analysis, we calculated linear (directional, ß), quadratic (stabilizing or disruptive, γ) and correlational (ρ) coefficients for DEEH and fat accumulation. We found significant, negative directional selection for DEEH (ßDEEH = - 0.58 ± 0.09), a positive value for fat accumulation (ßFAT = 0.34 ± 0.07), and positive correlational selection between both traits (ρDEEH × FAT = 0.24 ± 0.07). Then, individuals maximizing previous fat accumulation and minimizing DEEH were promoted by selection, which is visualized by a bi-variate selection surface estimated by generalized additive models. At the comparative level, results fall within the isometric allometry known for hibernation metabolic rate in mammals. Thus, by a combination of a non-invasive technique for body composition analysis and semi-natural enclosures, we were characterized the heterothermic fitness landscape in a semi-natural population of hibernators.

Individual diet specialisation in sparrows is driven by phenotypic plasticity in traits related to trade-offs in animal performance.

Individual diet specialisation (IS) is frequent in many animal taxa and affects population and community dynamics. The niche variation hypothesis (NVH) predicts that broader population niches should exhibit greater IS than populations with narrower niches, and most studies that examine the ecological factors driving IS focus on intraspecific competition. We show that phenotypic plasticity of traits associated with functional trade-offs is an important, but unrecognised mechanism that promotes and maintains IS. We measured nitrogen isotope (δ15 N) and digestive enzyme plasticity in four populations of sparrows (Zonotrichia capensis) to explore the relationship between IS and digestive plasticity. Our results show that phenotypic plasticity associated with functional trade-offs is related in a nonlinear fashion with the degree of IS and positively with population niche width. These findings are opposite to the NVH and suggest that among individual differences in diet can be maintained via acclimatisation and not necessarily require a genetic component.

The interplay between ambient temperature and salt intake affects oxidative status and immune responses in a ubiquitous Neotropical passerine, the rufous-collared sparrow.

Physiological traits associated with maintenance, growth, and reproduction demand a large amount of energy and thus directly influence an animal's energy budget, which is also regulated by environmental conditions. In this study, we evaluated the interplay between ambient temperature and salinity of drinking water on energy budgets and physiological responses in adult Rufous-collared sparrow (Zonotrichia capensis), an omnivorous passerine that is ubiquitous in Chile and inhabits a wide range of environments. We acclimated birds to 30 days at two ambient temperatures (27 °C and 17 °C) and drinking water salinity (200 mM NaCl and fresh water) conditions. We evaluated: 1) the aerobic scope and the activities of mitochondrial metabolic enzymes, 2) osmoregulatory parameters, 3) the skin-swelling immune response to an antigen, 4) oxidative status, and 5) the length of telomeres of red blood cells. Our results confirm that Z. capensis tolerates the chronic consumption of moderate levels of salt, maintaining body mass but increasing their basal metabolic rates consistent with expected osmoregulatory costs. Additionally, the factorial aerobic scope was higher in birds acclimated to fresh (tap) water at both 17° and 27 °C. Drinking water salinity and low ambient temperatures negatively impacted inflammatory response, and we observed an increase in lipid peroxidation and high levels of circulating antioxidants at low temperatures. Finally, telomere length was not affected by osmo- and thermoregulatory stress. Our results did not support the existence of an interplay between environmental temperature and drinking water salinity on most physiological and biochemical traits in Z. capensis, but the negative effect of these two factors on the inflammatory immune response suggests the existence of an energetic trade-off between biological functions that act in parallel to control immune function.

Contrasting seasonal and aseasonal environments across stages of the annual cycle in the rufous-collared sparrow, Zonotrichia capensis: Differences in endocrine function, proteome and body condition.

The timing and duration of life-history stages (LHSs) within the annual cycle can be affected by local environmental cues which are integrated through endocrine signalling mechanisms and changes in protein function. Most animals express a single LHS within a given period of the year because synchronous expression of LHSs is thought to be too costly energetically. However, in very rare and extremely stable conditions, breeding and moult have been observed to overlap extensively in rufous-collared sparrows (Zonotrichia capensis) living in valleys of the Atacama Desert-one of the most stable and aseasonal environments on Earth. To examine how LHS traits at different levels of organization are affected by environmental variability, we compared the temporal organization and duration of LHSs in populations in the Atacama Desert with those in the semiarid Fray Jorge National Park in the north of Chile-an extremely seasonal climate but with unpredictable droughts and heavy rainy seasons. We studied the effects of environmental variability on morphological variables related to body condition, endocrine traits and proteome. Birds living in the seasonal environment had a strict temporal division of LHSs, while birds living in the aseasonal environment failed to maintain a temporal division of LHSs resulting in direct overlap of breeding and moult. Further, higher circulating glucocorticoids and androgen concentrations were found in birds from seasonal compared to aseasonal populations. Despite these differences, body condition variables and protein expression were not related to the degree of seasonality but rather showed a strong relationship with hormone levels. These results suggest that animals adjust to their environment through changes in behavioural and endocrine traits and may be limited by less labile traits such as morphological variables or expression of specific proteins under certain circumstances. These data on free-living birds shed light on how different levels of life-history organization within an individual are linked to increasing environmental heterogeneity.

Intraspecific variation in exploratory behavior and elevational affinity in a widely distributed songbird.

Populations of the same species can vary substantially in their behavioral and morphometric traits when they are subject to different environmental pressures, which may lead to the development of different adaptive strategies. We quantified variation in exploratory behavior and morphometric traits among two rufous-collared sparrow populations that occur at low and high elevations in central Chile. Moreover, we used census and δ2H values of feather and blood to evaluate migration. We found that individual sparrows inhabiting high elevations were larger and showed more intense exploratory behavior in comparison with those that were captured at lower elevation. Moreover, we observed a steady decline in sparrow abundance during the winter and similar δ2H values for blood collected in the winter and summer at this site, which were significantly lower than blood Î´2H values observed at low elevation. This pattern suggests that individuals do not move long distances during winter, and likely they remain at similar elevations in refuge habitats. As predicted, our results support the existent of different adaptive strategies among populations of the same species, and suggest that the combination of behavioral, morphometric, and stable isotope data is a novel and robust integrative approach to assess differences in adaptation across environmental gradients.

Terrestrial birds in coastal environments: metabolic rate and oxidative status varies with the use of marine resources.

Life in saline environments represents a major physiological challenge for birds, particularly for passerines that lack nasal salt glands and hence are forced to live in environments that do not contain salty resources. Increased energy costs associated with increased salt intake, which in turn increases the production of reactive oxygen species, is likely a major selection pressure for why passerines are largely absent from brackish and marine environments. Here we measured basal metabolic rates (BMR) and oxidative status of free-ranging individuals of three species of Cinclodes, a group of passerine birds that inhabit marine and freshwater habitats in Chile. We used a combination of carbon, nitrogen, and hydrogen isotope data from metabolically active (blood) and inert (feathers) tissues to estimate seasonal changes in marine resource use and infer altitudinal migration. Contrary to our expectations, the consumption of marine resources did not result in higher BMR values and higher oxidative stress. Specifically, the marine specialist C. nigrofumosus had lower BMR than the other two species (C. fuscus and C. oustaleti), which seasonally switch between terrestrial and marine resources. C. fuscus had significantly higher total antioxidant capacity than the other two species (C. nigrofumosus and C. oustaleti) that consumed a relatively high proportion of marine resources. Nearly all studies examining the effects of salt consumption have focused on intraspecific acclimation via controlled experiments in the laboratory. The mixed results obtained from field- and lab-based studies reflect our poor understanding of the mechanistic link among hydric-salt balance, BMR, and oxidative stress in birds.

Is there an effect of environmental temperature on the response to an antigen and the metabolic rate in pups of the rodent Octodon degus?

Environmental temperature is a variable that influences all aspects of organisms, from physiological, e.g. immune function, and morphological traits to behavior. Recent studies have reported that environmental temperature modulates organisms' thermoregulatory capacity and immune response, suggesting that trade-offs must be made between thermoregulation and immune function. Despite this, studies that evaluate this trade-off in developing endotherms are scarce. The aim of this study was to evaluate the effects of environmental temperature experienced during development on the response to an antigen and its energetic costs in the precocial rodent Octodon degus. To accomplish this, we acclimated pups from birth to weaning at temperatures of 15°C and 30°C. At weaning, animals were inoculated with lipopolysaccharide (LPS) and cytokine interleukin-1ß levels, sickness behavior, changes in body temperature and basal metabolic rate, and body mass were measured. Our results showed that environmental temperature influences cytokine levels, body temperature, and some aspects of sickness behavior. Specifically, acclimatization at 30°C has a suppressive effect on the response to LPS, possibly due to a control to avoid overproduction of interleukin-1ß. Body mass and basal metabolic rate were not affected by environmental temperature experienced during development, but inoculation with LPS affected both variables. Our results suggest that ambient temperature may be a key factor that affects the response to an antigen in pups of O. degus; however, no evidence of a trade-off between thermoregulation and immune function was found here.

Testing the niche variation hypothesis in a community of passerine birds.

The niche variation hypothesis (NVH) predicts that populations with broader niches should exhibit greater between-individual diet variation or individual specialization (IS) relative to populations with narrower niches. Most studies that quantify population niche widths and associated levels of IS typically focus on a single or few species, but studies examining NVH in a phylogenetically informed comparative analysis among species are lacking. Here we use nitrogen isotope (δ15 N) analysis to measure population niche widths and IS in a single bird community composed of 12 passerine species representing different foraging guilds. We found support for the NVH at the interspecific level; species with broader population niche widths were comprised of more individual specialists. Moreover, our results suggest that this relationship is influenced by foraging guild; specifically, omnivores have higher degrees of IS for a given population niche width than insectivores. Finally, the levels of IS among passerine species, in contrast to population niche width, were associated with their relatedness, suggesting that the potential phylogenetic effect on the prevalence of IS is higher than previously recognized.

Early life experience drives short-term acclimation of metabolic and osmoregulatory traits in the leaf-eared mouse.

We studied the putative effect of early life experience on the physiological flexibility of metabolic and osmoregulatory traits in the leaf-eared mouse, Phyllotis darwini, an altricial rodent inhabiting seasonal Mediterranean environments. Adult individuals were collected in central Chile and maintained in breeding pairs. Pups were isolated after weaning and acclimated to different temperatures (cold or warm) and water availability (unrestricted and restricted) until adulthood. Subsequently, individuals were re-acclimated to the opposite treatment. Rodents reared in the warm and subjected to water restriction had lower basal metabolic rate (BMR), total evaporative water loss (TEWL) and body mass (Mb) compared with those developing in the cold treatment; nevertheless, individuals subjected to warm temperatures had greater relative medullary thickness (RMT) and urine concentrating ability (UCA). Cold-reared rodents re-acclimated to warm conditions exhibited physiological flexibility of metabolic traits; however, their osmoregulatory attributes did not vary. Conversely, warm-reared rodents re-acclimated to cold had reduced RMT and UCA, but the metabolic traits of these individuals did not change. These results suggest a trade-off between metabolic performance and renal capabilities that might hinder physiological acclimation. Our results support the hypothesis of ontogenetic dependence of short-term acclimation in osmoregulatory and metabolic traits in P. darwini.

Isotopic niches support the resource breadth hypothesis.

Because a broad spectrum of resource use allows species to persist in a wide range of habitat types, and thus permits them to occupy large geographical areas, and because broadly distributed species have access to more diverse resource bases, the resource breadth hypothesis posits that the diversity of resources used by organisms should be positively related with the extent of their geographic ranges. We investigated isotopic niche width in a small radiation of South American birds in the genus Cinclodes. We analysed feathers of 12 species of Cinclodes to test the isotopic version of the resource breadth hypothesis and to examine the correlation between isotopic niche breadth and morphology. We found a positive correlation between the widths of hydrogen and oxygen isotopic niches (which estimate breadth of elevational range) and widths of the carbon and nitrogen isotopic niches (which estimates the diversity of resources consumed, and hence of habitats used). We also found a positive correlation between broad isotopic niches and wing morphology. Our study not only supports the resource breadth hypothesis but it also highlights the usefulness of stable isotope analyses as tools in the exploration of ecological niches. It is an example of a macroecological application of stable isotopes. It also illustrates the importance of scientific collections in ecological studies.

Interplay between group size, huddling behavior and basal metabolism: an experimental approach in the social degu.

Mammals exposed to low temperatures increase their metabolic rate to maintain constant body temperature and thus compensate for heat loss. This high and costly energetic demand can be mitigated through thermoregulatory behavior such as social grouping or huddling, which helps to decrease metabolic rate as function of the numbers of individuals grouped. Sustained low temperatures in endothermic animals produce changes over time in rates of energy expenditure, by means of phenotypic plasticity. However, the putative modulating effect that huddling exerts on the flexibility of the basal metabolic rate (BMR) due to thermal acclimation remains unknown. We determined BMR values in Octodon degus, an endemic Chilean rodent, after being acclimated to either 15 or 30°C during 60 days, both alone and in groups of three and five individuals. At 15°C, BMR of huddling individuals was 40% lower than that of animals housed alone. Moreover, infrared thermography revealed a significant increase in local surface temperatures in huddled animals. Furthermore, individual thermal conductance was lower in individuals acclimated to 15°C than to 30°C, but no differences were observed between single and grouped animals. Our results indicate that huddling prevents an increase in BMR when animals are acclimated to cold conditions and that this effect is proportional to the number of animals grouped.

Living with voracious roommates: Factors that explain isotopic niche variation in a mixed colony of insectivorous bats.

Theory predicts that in resource-limited environments, coexisting species may overlap their niche dimensions but must differ in at least one to avoid competitive exclusion. Specifically, it has been suggested that the coexistence of competing species within a guild, could be sustained with mechanisms of resource partitioning, such as segregation along a trophic dimension. Among the most gregarious mammals are bats, which present diversification in their diet based on habitat choice and body size. Despite differences that could explain specialization in prey selection, there are insufficient studies that explore food overlap in mixed bat colonies and the factors that determine the selection of prey, both at intra- and inter-specific levels. To fill this gap, we analyzed the isotope signal (δ13C and δ15N) in feces collected in a mixed colony of Tadarida brasiliensis and Myotis chiloensis. To understand how several factors could influence these isotopic signals, intrinsic explanatory variables were analyzed, including body mass, body length, age, and sex. Also, extrinsic variables were analyzed, including monthly temporality and moonlight intensity. Our findings support age-dependent specialization in M. chiloensis, with a significant role of moonlight intensity and sex on δ15N. In T. brasiliensis, we identified a significant effect of size, sex, and ear length on δ15N. Our analysis indicates that both species of bats experience diverse degrees of overlap through austral summer months, affected by several factors that explain the variability in their fecal isotopic signals.

The effects of urban thermal heterogeneity and feather coloration on oxidative stress and metabolism of pigeons (Columba livia).

Urbanization stands out as a significant anthropogenic factor, exerting selective pressures on ecosystems and biotic components. A notable outcome of urbanization is thermal heterogeneity where the emergence of Urban Heat Islands is characterized by elevated air and surface temperatures compared to adjacent rural areas. Investigating the influence of thermal heterogeneity on urban animals could offer insights into how temperature variations can lead to phenotypic shifts. Urban pigeons (Columba livia) serve as an excellent model for studying urban thermal effects, given the melanism variations, which are associated with the pleiotropy of the melanocortin system. To examine the development of physiological plasticity in response to urban thermal variations, we conducted a study on pigeons in Santiago, Chile, during the rainy season. We assessed the influence of habitat on physiological traits related to metabolism and antioxidant capacities, which are theoretically affected by feather coloration. Our findings reveal that variations in melanism significantly impact pigeon physiology, affecting both antioxidant capacities and the mitochondrial activity of red blood cells. It was found that higher urban temperatures, from both the current sampling month and the prior sampling month (from CRU TS dataset), were negatively and strongly associated with lower antioxidant and metabolic activities. This suggests that elevated urban temperatures likely benefit the energetic budgets of pigeon populations and mitigate the negative effects of oxidative metabolism, with differential effects depending on feather colorations.

Osmoregulatory and metabolic costs of salt excretion in the Rufous-collared sparrow Zonotrichia capensis.

Recent experiments on shorebirds have demonstrated that maintaining an active osmoregulatory machinery is energetically expensive. This may, in part, explain diet and habitat selection in birds with salt glands. However little is known about the osmoregulatory costs in birds lacking functional salt glands. In these birds, osmotic work is done almost exclusively by the kidneys. We investigated the osmoregulatory cost in a bird species lacking functional salt glands, the passerine Zonotrichia capensis. After 20 days of acclimation to fresh water (FW) and salt water (200 mM NaCl, SW), SW birds tended to be heavier than FW birds. However, this difference was not statistically significant. Total basal metabolic rate was higher in SW birds as compared with FW birds. Renal and heart masses were also higher in the SW group. We also found greater medullary development and an increase in urine osmolality in the SW group. In spite of Z. capensis' ability to tolerate a moderate salt load in the laboratory, we hypothesize that increased cost of maintenance produced by salt consumption may significantly affect energy budget, dietary, and habitat choices in the field.

Prenatal caloric restriction adjusts the energy homeostasis and behavior in response to acute and chronic variations in food availability in adulthood.

Fetal metabolic programming produced by unfavorable prenatal nutritional conditions leads to the development of a disorder called "thrifty phenotype", which is associated with pathologies such as diabetes and obesity in adulthood. However, from an ecophysiological approach, few studies have addressed the development of thrifty phenotypes in terms of energy. This might represent an adaptive advantage against caloric deficiency conditions extending into adulthood. The objective of this study is to investigate the potential adaptive value of the thrifty phenotype expression through prenatal programming in a rodent model experiencing varying dietary conditions in different temporal contexts. To fill this gap, adult males of Mus musculus (BALB/C) from two maternal pregnancy groups were analyzed: control (ad libitum feeding) and caloric restriction from day 10 of gestation (70% restriction). Adult offspring of these groups were split further for two experiments: acute food deprivation and chronic caloric restriction at 60%. The acute food deprivation was performed for 24, 48 or 72 h while the caloric restriction regime was sustained for 20 days. For each experiment, morphological variables, such as body and organ mass, and gene expression related to lipid and carbohydrate metabolism from the liver and brain, were evaluated. In chronic caloric restriction, behavioral tests (open-field test and home-cage behavior) were performed. Our results indicate that under acute deprivation, the liver mass and triglyceride content remained unchanged in individuals subjected to prenatal restriction, in contrast to the reduction experienced by the control group. The latter is associated with the expression of the key genes involved in energy homeostasis (Pepck, Pparα/Pparγ), indicating a differential use of nutritional resources. In addition, thrifty animals, subjected to chronic caloric restriction, showed a severe reduction in locomotor and gluconeogenic activity, which is consistent with the regulatory role of Sirt1 and its downstream targets Mao and Pepck. Our results reveal that prenatal caloric restriction translates into a sparing metabolism in response to acute and chronic lack of food in adulthood.

Predicting the bioremediation potential of earthworms of different ecotypes through a multi-biomarker approach.

Earthworms are attracting the attention of bioremediation research because of their short-term impact on pollutant fate. However, earthworm-assisted bioremediation largely depends on the earthworm sensitivity to target pollutants and its metabolic capacity to break down contaminants. The most studied species in soil bioremediation has been Eisenia fetida, which inhabits the soil surface feeding on decomposing organic residues. Therefore, its bioremediation potential may be limited to organic matter-rich topsoil. We compared the detoxification potential against organophosphate (OP) pesticides of three earthworm species representative of the main ecotypes: epigeic, anecic, and endogeic. Selected biomarkers of pesticide detoxification (esterases, cytochrome P450-dependent monooxygenase, and glutathione S-transferase) and oxidative homeostasis (total antioxidant capacity, glutathione levels, and glutathione reductase [GR] and catalase activities) were measured in the muscle wall and gastrointestinal tract of E. fetida (epigeic), Lumbricus terrestris (anecic) and Aporrectodea caliginosa (endogeic). Our results show that L. terrestris was the most suitable species to bioremediate OP-contaminated soil for the following reasons: 1) Gut carboxylesterase (CbE) activity of L. terrestris was higher than that of E. fetida, whereas muscle CbE activity was more sensitivity to OP inhibition than that of E. fetida, which means a high capacity to inactivate the toxic oxon metabolites of OPs. 2) Muscle and gut phosphotriesterase activities were significantly higher in L. terrestris than in the other species. 3) Enzymatic (catalase and GR) and molecular mechanisms of free radical inactivation (glutathione) were 3- to 4-fold higher in L. terrestris concerning E. fetida and A. caliginosa, which reveals a higher potential to keep the cellular oxidative homeostasis against reactive metabolites formed during OP metabolism. Together with biological and ecological traits, these toxicological traits suggest L. terrestris a better candidate for soil bioremediation than epigeic earthworms.

Geographic variation in the altitudinal migration patterns, body size, oxidative status and exploratory behavior in a neotropical bird.

To cope with life in the mountains, populations of the same species can exhibit substantial variability in their altitudinal migration patterns and phenotypes in response to local weather conditions. Studying such variability can provide valuable insights into how local populations respond to environmental challenges, and this information can be useful for conservation efforts in mountain ecosystems. Here, we used δ2H values of feathers and blood to evaluate latitudinal variation in altitudinal migration patterns and its possible links with body size, oxidative status, and exploratory behavior in 72 individuals of rufous-collared sparrow (Zonotrichia capensis) that breed at low and high elevations in the center (~33°) and south (~38°) of Chile. Our results show that both altitudinal migration patterns and oxidative status were significantly influenced by the latitude of breeding sites, while exploratory behavior was associated with elevation. Notably, we found that fast-explorer birds inhabiting low elevations in central Chile displayed higher levels of oxidative damage than slow-explorer birds. These outcomes underscore the possibility of local adaptations in response to diverse local environmental conditions in the Andes. We discuss the implications of latitude, elevation, and environmental temperature in shaping the observed patterns and highlight the significance of identifying local adaptations in mountain birds for better predicting their response to climate change and other challenges stemming from anthropogenic activities.

Biochemical and osmoregulatory responses of the African clawed frog experimentally exposed to salt and pesticide.

Salinization and pollution are two main environmental stressors leading deterioration to water quality and degradation of aquatic ecosystems. Amphibians are a highly sensitive group of vertebrates to environmental disturbance of aquatic ecosystems. However, studies on the combined effect of salinization and pollution on the physiology of amphibians are limited. In this study, we measured the standard metabolic rate (SMR) and biochemical parameters of adult males of the invasive frog Xenopus laevis after 45 days of exposure to contrasting salinity environments (400 and 150 mOsm NaCl) with either 1.0 µg/L of the organophosphate pesticide chlorpyrifos (CPF) or pesticide-free medium. Our results revealed a decrease in SMR of animals exposed to the pesticide and in the ability to concentrate the plasma in animals exposed simultaneously to both stressors. The lack of ability to increase plasma concentration in animals exposed to both salt water and CPF, suggests that osmoregulatory response is decreased by pesticide exposure. In addition, we found an increase of liver citrate synthase activity in response to salt stress. Likewise, the liver acetylcholinesterase (AChE) activity decreased by 50% in frogs exposed to salt water and CPF and 40% in those exposed only to CPF, which suggest an additive effect of salinity on inhibition of AChE. Finally, oxidative stress increased as shown by the higher lipid peroxidation and concentration of aqueous peroxides found in the group exposed to salt water and pesticide. Thus, our results revealed that X. laevis physiology is compromised by salinization and pesticide exposure to both environmental stressors join.

Synergistic effects of pesticides and environmental variables on earthworm standard metabolic rate.

Endogeic earthworms such as Aporrectodea caliginosa play an essential role in the agroecosystems because of their continuous burrowing and feeding (geophagous) activity, which causes a profound impact on soil texture, organic matter decomposition, soil carbon storage, microbial activity, soil biodiversity, and nutrient cycling. Accordingly, endogeic earthworms are being proposed as suitable candidates for the ecotoxicity assessment of polluted soils. However, terrestrial ecotoxicology has little considered the interactive effects from pollutants and environmental variables (temperature, moisture). We acclimatized A. caliginosa for 90 days to two contrasting temperatures (10 °C and 20 °C) and moistures (25 % and 35 %, w/v) in soils contaminated with 20 mg kg-1 of chlorpyrifos to examine how these two climate change drivers may modulate the pesticide toxicity. We measured the inhibition of cholinesterase (ChE) activities as indicators of organophosphorus exposure, the standard metabolic rate as an integrative physiological biomarker, and the lipid peroxidation (TBARS) and the total antioxidant capacity (TAC) both as indicators of oxidative stress. The main results were: i) chlorpyrifos strongly inhibited ChE activity (>75 % of controls), demonstrating earthworm bioavailability and acute toxicity at the test concentration; 2) a 50 % mortality and loss of body weight (49 %) were found in the earthworms exposed to the most severe environmental conditions (20 °C, 25 %, and pesticide); 3) this latter experimental group displayed a high SMR, which was concomitant with an increase of the oxidative balance index (TBARS/TAC). We postulated that earthworms acclimatized to stressing environmental conditions experienced a higher pesticide-induced metabolic cost and physiological challenges imposed by adverse environmental conditions.