Indigo-dyed cellulose fibers and synthetic polymers in surface-feeding seabird chick regurgitates from the Gulf of Alaska.

We provide evidence of anthropogenic materials ingestion in seabirds from a remote oceanic area, using regurgitates obtained from black-legged kittiwake (Rissa tridactyla) chicks from Middleton Island (Gulf of Alaska, USA). By means of GPS tracking of breeding adults, we identified foraging grounds where anthropogenic materials were most likely ingested. They were mainly located within the continental shelf of the Gulf of Alaska and near the Alaskan coastline. Anthropogenic cellulose fibers showed a high prevalence (85 % occurrence), whereas synthetic polymers (in the micro- and mesoplastics dimensional range) were less frequent (20 %). Most fibers (60 %) were blue and we confirmed the presence of indigo-dyed cellulosic fibers, characteristic of denim fabrics. In terms of mass, contamination levels were 0.077 µg g-1 wet weight and 0.009 µg g-1 wet weight for anthropogenic microfibers and synthetic polymers, respectively. These results represent the only recent report of contamination by anthropogenic fibers in seabirds from the Gulf of Alaska.

A Chromosome-Level Reference Genome for the Black-Legged Kittiwake (Rissa tridactyla), a Declining Circumpolar Seabird.

Amidst the current biodiversity crisis, the availability of genomic resources for declining species can provide important insights into the factors driving population decline. In the early 1990s, the black-legged kittiwake (Rissa tridactyla), a pelagic gull widely distributed across the arctic, subarctic, and temperate zones, suffered a steep population decline following an abrupt warming of sea surface temperature across its distribution range and is currently listed as Vulnerable by the International Union for the Conservation of Nature. Kittiwakes have long been the focus for field studies of physiology, ecology, and ecotoxicology and are primary indicators of fluctuating ecological conditions in arctic and subarctic marine ecosystems. We present a high-quality chromosome-level reference genome and annotation for the black-legged kittiwake using a combination of Pacific Biosciences HiFi sequencing, Bionano optical maps, Hi-C reads, and RNA-Seq data. The final assembly spans 1.35 Gb across 32 chromosomes, with a scaffold N50 of 88.21 Mb and a BUSCO completeness of 97.4%. This genome assembly substantially improves the quality of a previous draft genome, showing an approximately 5× increase in contiguity and a more complete annotation. Using this new chromosome-level reference genome and three more chromosome-level assemblies of Charadriiformes, we uncover several lineage-specific chromosome fusions and fissions, but find no shared rearrangements, suggesting that interchromosomal rearrangements have been commonplace throughout the diversification of Charadriiformes. This new high-quality genome assembly will enable population genomic, transcriptomic, and phenotype-genotype association studies in a widely studied sentinel species, which may provide important insights into the impacts of global change on marine systems.

Beyond body condition: Experimental evidence that plasma metabolites improve nutritional state measurements in a free-living seabird.

The ability to efficiently measure the health and nutritional status of wild populations in situ is a valuable tool, as many methods of evaluating animal physiology do not occur in real-time, limiting the possibilities for direct intervention. This study investigates the use of blood plasma metabolite concentrations, measured via point-of-care devices or a simple plate reader assay, as indicators of nutritional state in free-living seabirds. We experimentally manipulated the energy expenditure of wild black-legged kittiwakes on Middleton Island, Alaska, and measured the plasma concentrations of glucose, cholesterol, B-hydroxybutyrate, and triglycerides throughout the breeding season, along with measures of body condition (size-corrected mass [SCM] and muscle depth). Supplemental feeding improved the nutritional state of kittiwakes by increasing feeding rate (higher glucose and triglycerides, lower cholesterol), and flight-handicapping caused a slight nutritional decline (lower glucose and triglycerides, higher cholesterol and B-hydroxybutyrate). Glucose and triglycerides were the best indicators of nutritional state when used alongside SCM, and improved upon commonly used metrics for measuring individual condition (i.e. SCM or mass alone). Metabolite concentrations varied across the breeding period, suggesting that the pre-laying stage, when feeding rates tend to be lower, was the most nutritionally challenging period for kittiwakes (low glucose, high cholesterol). Muscle depth also varied by treatment and breeding stage, but differed from other nutritional indices, suggesting that muscle depth is an indicator of exercise and activity level rather than nutrition. Here we demonstrate potential for the use of blood plasma metabolites measured via point-of-care devices as proxies for evaluating individual health, population health, and environmental food availability.

Sex-specific responses to GnRH challenge, but not food supply, in kittiwakes: Evidence for the "sensitivity to information" hypothesis.

Seasonal timing of breeding is usually considered to be triggered by endogenous responses linked to predictive cues (e.g., photoperiod) and supplementary cues that vary annually (e.g., food supply), but social cues are also important. Females may be more sensitive to supplementary cues because of their greater role in reproductive timing decisions, while males may only require predictive cues. We tested this hypothesis by food-supplementing female and male colonial seabirds (black-legged kittiwakes, Rissa tridactyla) during the pre-breeding season. We measured colony attendance via GPS devices, quantified pituitary and gonadal responses to gonadotropin releasing hormone (GnRH) challenge, and observed subsequent laying phenology. Food supplementation advanced laying phenology and increased colony attendance. While female pituitary responses to GnRH were consistent across the pre-breeding season, males showed a peak in pituitary sensitivity at approximately the same time that most females were initiating follicle development. The late peak in male pituitary response to GnRH questions a common assumption that males primarily rely on predictive cues (e.g., photoperiod) while females also rely on supplementary cues (e.g., food availability). Instead, male kittiwakes may integrate synchronising cues from their social environment to adjust their reproductive timing to coincide with female timing.

Lower nutritional state and foraging success in an Arctic seabird despite behaviorally flexible responses to environmental change.

The degree to which individuals adjust foraging behavior in response to environmental variability can impact foraging success, leading to downstream impacts on fitness and population dynamics. We examined the foraging flexibility, average daily energy expenditure, and foraging success of an ice-associated Arctic seabird, the thick-billed murre (Uria lomvia) in response to broad-scale environmental conditions at two different-sized, low Arctic colonies located <300 km apart. First, we compared foraging behavior (measured via GPS units), average daily energy expenditure (estimated from GPS derived activity budgets), and foraging success (nutritional state measured via nutritional biomarkers pre- and post- GPS deployment) of murres at two colonies, which differ greatly in size: 30,000 pairs breed on Coats Island, Nunavut, and 400,000 pairs breed on Digges Island, Nunavut. Second, we tested whether colony size within the same marine ecosystem altered foraging behavior in response to broad-scale environmental variability. Third, we tested whether environmentally induced foraging flexibility influenced the foraging success of murres. Murres at the larger colony foraged farther and longer but made fewer trips, resulting in a lower nutritional state and lower foraging success compared to birds at the smaller colony. Foraging behavior and foraging success varied in response to environmental variation, with murres at both colonies making longer, more distant foraging trips in high ice regimes during incubation, suggesting flexibility in responding to environmental variability. However, only birds at the larger colony showed this same flexibility during chick rearing. Foraging success at both colonies was higher during high ice regimes, suggesting greater prey availability. Overall, murres from the larger colony exhibited lower foraging success, and their foraging behavior showed stronger responses to changes in broad-scale conditions such as sea ice regime. Taken together, this suggests that larger Arctic seabird colonies have higher behavioral and demographic sensitivity to environmental change.

Ecological inference using data from accelerometers needs careful protocols.

Accelerometers in animal-attached tags are powerful tools in behavioural ecology, they can be used to determine behaviour and provide proxies for movement-based energy expenditure. Researchers are collecting and archiving data across systems, seasons and device types. However, using data repositories to draw ecological inference requires a good understanding of the error introduced according to sensor type and position on the study animal and protocols for error assessment and minimisation.Using laboratory trials, we examine the absolute accuracy of tri-axial accelerometers and determine how inaccuracies impact measurements of dynamic body acceleration (DBA), a proxy for energy expenditure, in human participants. We then examine how tag type and placement affect the acceleration signal in birds, using pigeons Columba livia flying in a wind tunnel, with tags mounted simultaneously in two positions, and back- and tail-mounted tags deployed on wild kittiwakes Rissa tridactyla. Finally, we present a case study where two generations of tag were deployed using different attachment procedures on red-tailed tropicbirds Phaethon rubricauda foraging in different seasons.Bench tests showed that individual acceleration axes required a two-level correction to eliminate measurement error. This resulted in DBA differences of up to 5% between calibrated and uncalibrated tags for humans walking at a range of speeds. Device position was associated with greater variation in DBA, with upper and lower back-mounted tags varying by 9% in pigeons, and tail- and back-mounted tags varying by 13% in kittiwakes. The tropicbird study highlighted the difficulties of attributing changes in signal amplitude to a single factor when confounding influences tend to covary, as DBA varied by 25% between seasons.Accelerometer accuracy, tag placement and attachment critically affect the signal amplitude and thereby the ability of the system to detect biologically meaningful phenomena. We propose a simple method to calibrate accelerometers that can be executed under field conditions. This should be used prior to deployments and archived with resulting data. We also suggest a way that researchers can assess accuracy in previously collected data, and caution that variable tag placement and attachment can increase sensor noise and even generate trends that have no biological meaning.

The role of wingbeat frequency and amplitude in flight power.

Body-mounted accelerometers provide a new prospect for estimating power use in flying birds, as the signal varies with the two major kinematic determinants of aerodynamic power: wingbeat frequency and amplitude. Yet wingbeat frequency is sometimes used as a proxy for power output in isolation. There is, therefore, a need to understand which kinematic parameter birds vary and whether this is predicted by flight mode (e.g. accelerating, ascending/descending flight), speed or morphology. We investigate this using high-frequency acceleration data from (i) 14 species flying in the wild, (ii) two species flying in controlled conditions in a wind tunnel and (iii) a review of experimental and field studies. While wingbeat frequency and amplitude were positively correlated, R2 values were generally low, supporting the idea that parameters can vary independently. Indeed, birds were more likely to modulate wingbeat amplitude for more energy-demanding flight modes, including climbing and take-off. Nonetheless, the striking variability, even within species and flight types, highlights the complexity of describing the kinematic relationships, which appear sensitive to both the biological and physical context. Notwithstanding this, acceleration metrics that incorporate both kinematic parameters should be more robust proxies for power than wingbeat frequency alone.

Food supply and individual quality influence seabird energy expenditure and reproductive success.

Breeding animals trade off maximizing energy output to increase their number of offspring with conserving energy to ensure their own survival, leading to an energetic ceiling influenced by external, environmental factors or by internal, physiological factors. We examined whether internal or external factors limited energy expenditure by supplementally feeding breeding black-legged kittiwakes varying in individual quality, based on earlier work that defined late breeders as low-quality and early breeders as high-quality individuals. We tested whether energy expenditure increased when food availability decreased in both low- and high-quality birds; we predicted this would only occur in high-quality individuals capable of sustaining high levels of energy expenditure. Here, we find that food-supplemented birds expended less energy than control birds because they spent more time at the colony. However, foraging trips of food-supplemented birds were only slightly shorter than control birds, implying that food-supplemented birds were limited by food availability at sea similarly to control birds. Late breeders expended less energy, suggesting that low-quality individuals may not intake the energy necessary for sustaining high-energy output. Food-supplemented birds had more offspring than control birds, but offspring number did not influence energy expenditure, supporting the idea that the birds reached an energy ceiling. Males and lighter birds expended more energy, possibly compensating for relatively higher energy intake. Chick-rearing birds were working near their maximum, with highest levels of expenditure for early-laying (high-quality) individuals foraging at sea. Due to fluctuating marine environments, kittiwakes may be forced to change their foraging behaviors to maintain the balance between reproduction and survival.

Resting costs too: the relative importance of active and resting energy expenditure in a sub-arctic seabird.

Breeding is costly for many animals, including birds that must deliver food to a central place (i.e. nest). Measuring energy expenditure throughout the breeding season can provide valuable insight into physiological limitations by highlighting periods of high demand, and ultimately allows improvement of conservation strategies. However, quantifying energy expenditure in wildlife can be challenging, as existing methods do not measure both active (e.g. foraging) and resting energy costs across short and long time scales. Here, we developed a novel method for comparing active and resting costs in 66 pre-breeding and breeding seabirds (black-legged kittiwakes, Rissa tridactyla) by combining accelerometry and triiodothyronine (T3) as proxies for active and resting costs, respectively. Active energy costs were higher during incubation (P=0.0004) and chick rearing (P<0.0001) than during pre-laying, because of an increase in the time spent in flight of 11% (P=0.0005) and 15% (P<0.0001), respectively. Levels of T3, reflecting resting costs, peaked marginally during incubation with a mean (±s.d.) concentration of 4.71±1.97 pg ml-1 in comparison to 2.66±1.30 pg ml-1 during pre-laying (P=0.05) and 3.16±2.85 pg ml-1 during chick rearing (P=0.11). Thus, although chick rearing is often assumed to be the costliest breeding stage by multiple studies, our results suggest that incubation could be more costly as a result of high resting costs. We highlight the importance of accounting for both active and resting costs when assessing energy expenditure.

Natal experience and pre-breeding environmental conditions affect lay date plasticity in Savannah Sparrows.

Phenotypic plasticity allows organisms to adjust the timing of life-history events in response to environmental and demographic conditions. Shifts by individuals in the timing of breeding with respect to variation in age and temperature are well documented in nature, and these changes are known to scale to affect population dynamics. However, relatively little is known about how organisms alter phenology in response to other demographic and environmental factors. We investigated how pre-breeding temperature, breeding population density, age, and rainfall in the first month of life influenced the timing and plasticity of lay date in a population of Savannah Sparrows (Passerculus sandwichensis) monitored over 33 yr (1987-2019). Females that experienced warmer pre-breeding temperatures tended to lay eggs earlier, as did older females, but breeding population density had no effect on lay date. Natal precipitation interacted with age to influence lay date plasticity, with females that experienced high precipitation levels as nestlings advancing lay dates more strongly over the course of their lives. We also found evidence for varied pace of life; females that experienced high natal precipitation had shorter lifespans and reduced fecundity, but more nesting attempts over their lifetimes. Rainfall during the nestling period increased through time, while population density and fecundity declined, suggesting that increased precipitation on the breeding grounds may be detrimental to breeding females and ultimately the viability of the population as a whole. Our results suggest that females adjust their laying date in response to pre-breeding temperature, and as they age, while presenting new evidence that environmental conditions during the natal period can affect phenological plasticity and generate downstream, population-level effects.

Geolocators link marine mercury with levels in wild seabirds throughout their annual cycle: Consequences for trans-ecosystem biotransport.

Seabirds are widely used as indicators of marine pollution, including mercury (Hg), because they track contaminant levels across space and time. However, many seabirds are migratory, and it is difficult to understand the timing and location of their Hg accumulation. Seabirds may obtain Hg thousands of kilometers away, during their non-breeding period, and deposit that Hg into their terrestrial breeding colonies. We predicted that Hg concentration in rectrices reflects exposure during the previous breeding season, in body feathers reflects non-breeding exposure, and in blood collected during breeding reflects exposure during current breeding. To test this hypothesis, we measured total Hg concentration in these three tissues, which reflect different timepoints during the annual cycle of rhinoceros auklets (Cerorhinca monocerata) breeding on both sides of the North Pacific (Middleton Island in Alaska and Teuri Island in Hokkaido), and tracked their wintering movement patterns with biologging devices. We (i) identify the wintering patterns of both populations, (ii) examine Hg levels in different tissues representing exposure at different time periods, (iii) test how environmental Hg exposure during the non-breeding season affects bird contamination, and (iv) assess whether variation in Hg levels during the non-breeding season influences levels accumulated in terrestrial plants. Individuals from both populations followed a figure-eight looping migration pattern. We confirm the existence of a pathway from environmental Hg to plant roots via avian tissues, as Hg concentrations were higher in plants within the auklet colonies than at control sites. Hg concentrations of breast feathers were higher in Alaskan than in Japanese auklets, but Hg concentrations in rectrices and blood were similar. Moreover, we found evidence that tissues with different turnover rates could record local anthropogenic Hg emission rates of areas visited during winter. In conclusion, Hg was transported across thousands of kilometers by seabirds and transferred to local plants.

The effects of food supply on reproductive hormones and timing of reproduction in an income-breeding seabird.

Current food supply is a major driver of timing of breeding in income-breeding animals, likely because increased net energy balance directly increases reproductive hormones and advances breeding. In capital breeders, increased net energy balance increases energy reserves, which eventually leads to improved reproductive readiness and earlier breeding. To test the hypothesis that phenology of income-breeding birds is independent of energy reserves, we conducted an experiment on food-supplemented ("fed") and control female black-legged kittiwakes (Rissa tridactyla). We temporarily increased energy costs (via weight handicap) in a 2 × 2 design (fed/unfed; handicapped/unhandicapped) during the pre-laying period and observed movement via GPS-accelerometry. We measured body mass, baseline hormones (corticosterone; luteinising hormone) before and after handicap manipulation, and conducted a gonadotropin-releasing hormone challenge. Females from all treatment groups foraged in similar areas, implying that individuals could adjust time spent foraging, but had low flexibility to adjust foraging distance. Consistent with the idea that income breeders do not accumulate reserves in response to increased food supply, fed birds remained within an energy ceiling by reducing time foraging instead of increasing energy reserves. Moreover, body mass remained constant until the onset of follicle development 20 days prior to laying regardless of feeding or handicap, implying that females were using a 'lean and fit' approach to body mass rather than accumulating lipid reserves for breeding. Increased food supply advanced endocrine and laying phenology and altered interactions between the hypothalamic-pituitary-adrenal axis and the hypothalamic-pituitary-gonadal axis, but higher energy costs (handicap) had little effect. Consistent with our hypothesis, increased food supply (but not net energy balance) advanced endocrine and laying phenology in income-breeding birds without any impact on energy reserves.

Accelerometry predicts muscle ultrastructure and flight capabilities in a wild bird.

Muscle ultrastructure is closely linked with athletic performance in humans and lab animals, and presumably plays an important role in the movement ecology of wild animals. Movement is critical for wild animals to forage, escape predators and reproduce. However, little evidence directly links muscle condition to locomotion in the wild. We used GPS-accelerometers to examine flight behaviour and muscle biopsies to assess muscle ultrastructure in breeding black-legged kittiwakes (Rissa tridactyla). Biopsied kittiwakes showed similar reproductive success and subsequent over-winter survival to non-biopsied kittiwakes, suggesting that our study method did not greatly impact foraging ability. Muscle fibre diameter was negatively associated with wing beat frequency, likely because larger muscle fibres facilitate powered flight. The number of nuclei per fibre was positively associated with average air speed, likely because higher power output needed by faster-flying birds required plasticity for muscle fibre recruitment. These results suggest the potential for flight behaviour to predict muscle ultrastructure.

Increased summer food supply decreases non-breeding movement in black-legged kittiwakes.

Individual condition at one stage of the annual cycle is expected to influence behaviour during subsequent stages, yet experimental evidence of food-mediated carry-over effects is scarce. We used a food supplementation experiment to test the effects of food supply during the breeding season on migration phenology and non-breeding behaviour. We provided an unlimited supply of fish to black-legged kittiwakes (Rissa tridactyla) during their breeding season on Middleton Island, Alaska, monitored reproductive phenology and breeding success, and used light-level geolocation to observe non-breeding behaviour. Among successful breeders, fed kittiwakes departed the colony earlier than unfed controls. Fed kittiwakes travelled less than controls during the breeding season, contracting their non-breeding range. Our results demonstrate that food supply during the breeding season affects non-breeding phenology, movement and distribution, providing a potential behavioural mechanism underlying observed survival costs of reproduction.

Point-of-care blood analyzers measure the nutritional state of eighteen free-living bird species.

Point-of-care devices offer the potential to democratize a suite of physiological endpoints and assess the nutritional state of wild animals through plasma metabolite profiling. Measurements of plasma metabolites typically occur on frozen tissue in the laboratory, thus dissociating measurements from field observations. Point-of-care devices, widely used in veterinary and human medicine, provide rapid results (seconds or minutes) allowing in situ measurements of wild animals in remote areas without the need for access to freezers. Using point-of-care devices, we measured glucose, triglyceride, cholesterol and ß-hydroxybutyrate levels in plasma from 18 wild bird species spanning nine families and three orders. The values from six different point-of-care devices correlated strongly with one another, and with traditional laboratory measurements from stored plasma (R2 = 0.70-0.90). Although POC devices provided accurate relative values in wild birds, absolute values varied from laboratory measurements by up to 50% illustrating the need for calibration equations. Furthermore, three case studies showed the potential for point-of-care devices at research stations where participants do not have access to a lab and sample preservation is difficult: (i) at a remote seabird colony, birds that were provided with supplemental food had higher levels of glucose and lower ß-hydroxybutyrate and cholesterol levels than unfed birds, suggesting they were in a better nutritional state; (ii) at a migration monitoring station, levels of triglycerides of two migratory songbirds increased with time of day, implying that they were fattening during stopover; and (iii) for diving seabirds, individuals that worked harder (shorter surface intervals) had higher glucose and lower ß-hydroxybutyrate implying that nutritional state is an index of foraging effort and success. We demonstrate that point-of-care devices, once validated, can provide accurate measurements of the nutritional state of wild birds. Such real-time measurements can aid in ecological research and monitoring, care of wildlife at rehabilitation centres, and in veterinary medicine of exotics.

Huffin' and puffin: seabirds use large bills to dissipate heat from energetically demanding flight.

Endothermic animals regulate body temperature by balancing metabolic heat production and heat exchange with the environment. Heat dissipation is especially important during and immediately after demanding activities such as flapping flight, the most energetically expensive mode of locomotion. As uninsulated appendages, bird bills present a potential avenue for efficient heat dissipation. Puffins possess large bills and are members of the bird family with the highest known flight cost. Here, we used infrared thermography to test whether wild tufted puffins (Fratercula cirrhata) use their bills to dissipate excess heat after energetically expensive flight. Both bill surface temperature and the proportion of total heat exchange occurring at the bill decreased with time since landing, suggesting that bills are used to dissipate excess metabolic heat. We propose that bill size in puffins may be shaped by opposing selective pressures that include dissipating heat after flight and conserving heat in cold air and water temperatures.

Muscle fiber structure in an aging long-lived seabird, the black-legged kittiwake (Rissa tridactyla).

Many long-lived animals do not appear to show classic signs of aging, perhaps because they show negligible senescence until dying from "catastrophic" mortality. Muscle senescence is seldom examined in wild animals, yet decline in muscle function is one of the first signs of aging in many lab animals and humans. Seabirds are an excellent study system for physiological implications of aging because they are long-lived animals that actively forage and reproduce in the wild. Here, we examined linkages between pectoralis muscle fiber structure and age in black-legged kittiwakes (Rissa tridactyla). Pectoralis muscle is the largest organ complex in birds, and responsible for flight and shivering. We obtained and fixed biopsies from wild black-legged kittiwakes of known age. We then measured muscle fiber diameter, myonuclear domain and capillaries per fiber area among birds of differing ages. All muscle parameters were independent of age. Number of nuclei per mm of fiber showed a positive correlation with muscle fiber cross-sectional area, and myonuclear domain increased with muscle fiber diameter. Thus, as muscle fibers increased in size, they may not have recruited satellite cells, increasing the protein turnover load per nuclei. We conclude that senescence in a long-lived bird with an active lifestyle, does not entail mammalian-like changes in muscle structure.

Reduced reproductive performance associated with warmer ambient temperatures during incubation in a winter-breeding, food-storing passerine.

Timing of reproduction can influence individual fitness whereby early breeders tend to have higher reproductive success than late breeders. However, the fitness consequences of timing of breeding may also be influenced by environmental conditions after the commencement of breeding. We tested whether ambient temperatures during the incubation and early nestling periods modulated the effect of laying date on brood size and dominant juvenile survival in gray jays (Perisoreus canadensis), a sedentary boreal species whose late winter nesting depends, in part, on caches of perishable food. Previous evidence has suggested that warmer temperatures degrade the quality of these food hoards, and we asked whether warmer ambient temperatures during the incubation and early nestling periods would be associated with smaller brood sizes and lower summer survival of dominant juveniles. We used 38 years of data from a range-edge population of gray jays in Algonquin Provincial Park, Ontario, where the population has declined over 50% since the study began. Consistent with the "hoard-rot" hypothesis, we found that cold temperatures during incubation were associated with larger brood sizes in later breeding attempts, but temperatures had little effect on brood size for females breeding early in the season. This is the first evidence that laying date and temperature during incubation interactively influence brood size in any bird species. We did not find evidence that ambient temperatures during the incubation period or early part of the nestling period influenced summer survival of dominant juveniles. Our findings provide evidence that warming temperatures are associated with some aspects of reduced reproductive performance in a species that is reliant on cold temperatures to store perishable food caches, some of which are later consumed during the reproductive period.