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.

Ecologically driven differences in individual diet specialization across three populations of Guiana dolphin.

Populations usually considered foraging generalists may include specialized individuals that feed on a restricted subset of the prey spectrum consumed by the population. By analyzing the time series of δ13C and δ15N values in sequential growth layer groups within tooth dentin, we measured population- and individual-level variation in resource use of three populations of Guiana dolphins (Sotalia guianensis)-Caravelas River, Babitonga Bay, and Norte Bay-along a latitudinal gradient in the southwestern Atlantic Ocean. We show that the Guiana dolphin at Caravelas River is a generalist population consisting of individual dietary specialists, likely due to the absence of other resident dolphin populations thus allowing individuals to target prey across a wide range of habitats. The Babitonga Bay population is also composed of individual specialists potentially due to the selective foraging behavior of some individuals on high-quality prey sources within and near the bay. In contrast, the Norte Bay population comprises individual generalists, which likely reflects its distinctive cohesive social organization, coexistence with two other dolphin species, and an opportunistic foraging strategy in response to resource fluctuations inherent to the southern limit of the species distribution. Although the Guiana dolphin is generally considered to be a dietary generalist at the population level, our findings reveal that the total niche width of populations and the degree of individual diet specialization are highly context dependent, suggesting dietary plasticity that may be related to a latitudinal gradient in resource availability and environmental conditions.

Triple Oxygen Isotope Measurements (Δ'17O) of Body Water Reflect Water Intake, Metabolism, and δ18O of Ingested Water in Passerines.

Understanding physiological traits and ecological conditions that influence a species reliance on metabolic water is critical to creating accurate physiological models that can assess their ability to adapt to environmental perturbations (e.g., drought) that impact water availability. However, relatively few studies have examined variation in the sources of water animals use to maintain water balance, and even fewer have focused on the role of metabolic water. A key reason is methodological limitations. Here, we applied a new method that measures the triple oxygen isotopic composition of a single blood sample to estimate the contribution of metabolic water to the body water pool of three passerine species. This approach relies on Δ'17O, defined as the residual from the tight linear correlation that naturally exists between δ17O and δ18O values. Importantly, Δ'17O is relatively insensitive to key fractionation processes, such as Rayleigh distillation in the water cycle that have hindered previous isotope-based assessments of animal water balance. We evaluated the effects of changes in metabolic rate and water intake on Δ'17O values of captive rufous-collared sparrows (Zonotrichia capensis) and two invertivorous passerine species in the genus Cinclodes from the field. As predicted, colder acclimation temperatures induced increases in metabolic rate, decreases in water intake, and increases in the contribution of metabolic water to the body water pool of Z. capensis, causing a consistent change in Δ'17O. Measurement of Δ'17O also provides an estimate of the δ18O composition of ingested pre-formed (drinking/food) water. Estimated δ18O values of drinking/food water for captive Z. capensis were ~ -11‰, which is consistent with that of tap water in Santiago, Chile. In contrast, δ18O values of drinking/food water ingested by wild-caught Cinclodes were similar to that of seawater, which is consistent with their reliance on marine resources. Our results confirm the utility of this method for quantifying the relative contribution of metabolic versus pre-formed drinking/food water to the body water pool in birds.

Intraspecific variation and energy channel coupling within a Chilean kelp forest.

The widespread importance of variable types of primary production, or energy channels, to consumer communities has become increasingly apparent. However, the mechanisms underlying this "multichannel" feeding remain poorly understood, especially for aquatic ecosystems that pose unique logistical constraints given the diversity of potential energy channels. Here, we use bulk tissue isotopic analysis along with carbon isotope (δ13 C) analysis of individual amino acids to characterize the relative contribution of pelagic and benthic energy sources to a kelp forest consumer community in northern Chile. We measured bulk tissue δ13 C and δ15 N for >120 samples; of these we analyzed δ13 C values of six essential amino acids (EAA) from nine primary producer groups (n = 41) and 11 representative nearshore consumer taxa (n = 56). Using EAA δ13 C data, we employed linear discriminant analysis (LDA) to assess how distinct EAA δ13 C values were between local pelagic (phytoplankton/particulate organic matter), and benthic (kelps, red algae, and green algae) endmembers. With this model, we were able to correctly classify nearly 90% of producer samples to their original groupings, a significant improvement on traditional bulk isotopic analysis. With this EAA isotopic library, we then generated probability distributions for the most important sources of production for each individual consumer and species using a bootstrap-resampling LDA approach. We found evidence for multichannel feeding within the community at the species level. Invertebrates tended to focus on either pelagic or benthic energy, deriving 13-67% of their EAA from pelagic sources. In contrast, mobile (fish) taxa at higher trophic levels used more equal proportions of each channel, ranging from 19% to 47% pelagically derived energy. Within a taxon, multichannel feeding was a result of specialization among individuals in energy channel usage, with 37 of 56 individual consumers estimated to derive >80% of their EAA from a single channel. Our study reveals how a cutting-edge isotopic technique can characterize the dynamics of energy flow in coastal food webs, a topic that has historically been difficult to address. More broadly, our work provides a mechanism as to how multichannel feeding may occur in nearshore communities, and we suggest this pattern be investigated in additional ecosystems.

Latitudinal patterns in the diet of Andean condor (Vultur gryphus) in Chile: Contrasting environments influencing feeding behavior.

Human-dominated environments alter the availability and quality of resources for many species, especially for scavengers that have large home ranges and plastic foraging behaviors that enable them to exploit novel resources. Along the western slope of the Andes, the modification of natural landscapes have resulted in significant declines in native prey, the introduction of non-native species, and an increase in the availability of anthropogenic resources. These factors have likely influenced the resources available to Andean condors (Vultur gryphus), however, data are lacking as to how condor's diet vary along their large latitudinal range. We evaluated differences in Andean condor diet along a ~2500 km latitudinal gradient in Chile from the heavily modified Central zone (32-34°S) to the more pristine Austral zone (44-56°S). We assessed diet composition through the identification of prey remains in condor pellets, and carbon and nitrogen isotope analysis of condor feathers and their primary prey identified from pellet analysis. Our results identified medium- and large-bodied domesticated mammals (ungulates) and introduced exotic species (lagomorphs) as common prey across the study area. Condors from the Central zone had the largest isotopic niche width, probably related to consumption of anthropogenic resources with distinctly high carbon isotope values indicative of C4-based foods likely acquired from landfills or corn-fed livestock. Isotopic niches for condors from the Southern and Austral zones almost completely overlapped. Andean condor diet is strongly influenced by local conditions determining differential access to prey sources. The high dependence of Andean condors on livestock across a large geographical area, and landfills in more (sub)urban areas, may help stabilize their populations via anthropogenic resources subsidies. Long-term dependence on such resources, however, may have health costs including contaminant exposure and greater mortality risk. These data will help identify potential threats related to resource availability and use, and better inform management and conservation decisions.

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.

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.

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.

Why are tropical mountain passes "low" for some species? Genetic and stable-isotope tests for differentiation, migration and expansion in elevational generalist songbirds.

Most tropical bird species have narrow elevational ranges, likely reflecting climatic specialization. This is consistent with Janzen's Rule, the tendency for mountain passes to be effectively "higher" in the tropics. Hence, those few tropical species that occur across broad elevational gradients (elevational generalists) represent a contradiction to Janzen's Rule. Here, we aim to address the following questions. Are elevational generalists being sundered by diversifying selection along the gradient? Does elevational movement cause these species to resist diversification or specialization? Have they recently expanded, suggesting that elevational generalism is short-lived in geological time? To answer these questions, we tested for differentiation, movement and expansion in four elevational generalist songbird species on the Andean west slope. We used morphology and mtDNA to test for genetic differentiation between high- and low-elevation populations. To test for elevational movements, we measured hydrogen isotope (δ2 H) values of metabolically inert feathers and metabolically active liver. Morphology differed for House Wren (Troglodytes aedon) and Hooded Siskin (Spinus magellanicus), but not for Cinereous Conebill (Conirostrum cinereum) and Rufous-collared Sparrow (Zonotrichia capensis) respectively. mtDNA was structured by elevation only in Z. capensis. δ2 H data indicated elevational movements by two tree- and shrub-foraging species with moderate-to-high vagility (C. cinereum and S. magellanicus), and sedentary behaviour by two terrestrial-foraging species with low-to-moderate vagility (T. aedon and Z. capensis). In S. magellanicus, elevational movements and lack of mtDNA structure contrast with striking morphological divergence, suggesting strong diversifying selection on body proportions across the c. 50 km gradient. All species except C. cinereum exhibited mtDNA haplotype variation consistent with recent population expansion across the elevational gradient, potentially concurrent with Holocene anthropogenic habitat conversion for agriculture. In different ways, each species defies the tendency for tropical birds to have long-term stable distributions and sedentary habits. We conclude that tropical elevational generalism is rare due to evolutionary instability.

Coping with Salt Water Habitats: Metabolic and Oxidative Responses to Salt Intake in the Rufous-Collared Sparrow.

Many physiological adjustments occur in response to salt intake in several marine taxa, which manifest at different scales from changes in the concentration of individual molecules to physical traits of whole organisms. Little is known about the influence of salinity on the distribution, physiological performance, and ecology of passerines; specifically, the impact of drinking water salinity on the oxidative status of birds has been largely ignored. In this study, we evaluated whether experimental variations in the salt intake of a widely-distributed passerine (Zontotrichia capensis) could generate differences in basal (BMR) and maximum metabolic rates (Msum), as well as affect metabolic enzyme activity and oxidative status. We measured rates of energy expenditure of birds after 30-d acclimation to drink salt (SW) or tap (fresh) water (TW) and assessed changes in the activity of mitochondrial enzymes (cytochrome c oxidase and citrate synthase) in skeletal muscle, heart, and kidney. Finally, we evaluated the oxidative status of bird tissues by means of total antioxidant capacity (TAC) and superoxide dismutase activities and lipid oxidative damage (Malondialdehyde, MDA). The results revealed a significant increase in BMR but not Msum, which resulted in a reduction in factorial aerobic scope in SW- vs. TW-acclimated birds. These changes were paralleled with increased kidney and intestine masses and catabolic activities in tissues, especially in pectoralis muscle. We also found that TAC and MDA concentrations were ~120 and ~400% higher, respectively in the liver of animals acclimated to the SW- vs. TW-treatment. Our study is the first to document changes in the oxidative status in birds that persistently drink saltwater, and shows that they undergo several physiological adjustments that range that range in scale from biochemical capacities (e.g., TAC and MDA) to whole organism traits (e.g., metabolic rates). We propose that the physiological changes observed in Z. capensis acclimated to saltwater could be common phenomena in birds and likely explain selection of prey containing little salt and habitats associated with low salinity.

Estimating blue whale skin isotopic incorporation rates and baleen growth rates: Implications for assessing diet and movement patterns in mysticetes.

Stable isotope analysis in mysticete skin and baleen plates has been repeatedly used to assess diet and movement patterns. Accurate interpretation of isotope data depends on understanding isotopic incorporation rates for metabolically active tissues and growth rates for metabolically inert tissues. The aim of this research was to estimate isotopic incorporation rates in blue whale skin and baleen growth rates by using natural gradients in baseline isotope values between oceanic regions. Nitrogen (δ15N) and carbon (δ13C) isotope values of blue whale skin and potential prey were analyzed from three foraging zones (Gulf of California, California Current System, and Costa Rica Dome) in the northeast Pacific from 1996-2015. We also measured δ15N and δ13C values along the lengths of baleen plates collected from six blue whales stranded in the 1980s and 2000s. Skin was separated into three strata: basale, externum, and sloughed skin. A mean (±SD) skin isotopic incorporation rate of 163±91 days was estimated by fitting a generalized additive model of the seasonal trend in δ15N values of skin strata collected in the Gulf of California and the California Current System. A mean (±SD) baleen growth rate of 15.5±2.2 cm y-1 was estimated by using seasonal oscillations in δ15N values from three whales. These oscillations also showed that individual whales have a high fidelity to distinct foraging zones in the northeast Pacific across years. The absence of oscillations in δ15N values of baleen sub-samples from three male whales suggests these individuals remained within a specific zone for several years prior to death. δ13C values of both whale tissues (skin and baleen) and potential prey were not distinct among foraging zones. Our results highlight the importance of considering tissue isotopic incorporation and growth rates when studying migratory mysticetes and provide new insights into the individual movement strategies of blue whales.

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.

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.

Using Satellite Tracking and Isotopic Information to Characterize the Impact of South American Sea Lions on Salmonid Aquaculture in Southern Chile.

Apex marine predators alter their foraging behavior in response to spatial and/or seasonal changes in natural prey distribution and abundance. However, few studies have identified the impacts of aquaculture that represents a spatially and temporally predictable and abundant resource on their foraging behavior. Using satellite telemetry and stable isotope analysis we examined the degree of spatial overlap between the South American sea lion (SASL) and salmon farms, and quantify the amount of native prey versus farmed salmonids in SASL diets. We instrumented eight SASL individuals with SRDL-GPS tags. Vibrissae, hair and skin samples were collected for δ13C and δ15N analyses from five of the tagged individuals and from four males captured in a haul-out located adjacent to salmon farms. Tracking results showed that almost all the foraging areas of SASL are within close proximity to salmon farms. The most important prey for the individuals analyzed was farmed salmonids, with an estimated median (±SD) contribution of 19.7 ± 13.5‰ and 15.3 ± 9.6‰ for hair and skin, respectively. Using vibrissae as a temporal record of diet for each individual, we observed a remarkable switch in diet composition in two SASL, from farmed salmonids to pelagic fishes, which coincided with the decrease of salmon production due to the infectious salmon anemia virus that affected salmon farms in Chile at the end of 2008. Our study demonstrates the usefulness of integrating stable isotope derived dietary data with movement patterns to characterize the impacts of a non-native prey on the foraging ecology of an apex marine predator, providing important applied implications in situations where interactions between aquaculture and wildlife are common.