Calypte Anna Research Articles

Geographical and ecological allopatry effects on niche change in two sister species pairs of hummingbirds in western North America

Here, we explored how variations in the allopatric speciation scenario, specifically ecological vs. vicariant allopatry, relate to climatic niche change in sister species. We selected two sister species pairs of North American hummingbirds (Calypte anna, C. costae, Basilinna leucotis, B. xantusii) that diverged 2.5 and 3.6 million years ago, under ecological (arid climate tendency during Pliocene) and vicariant (Baja California peninsula separation) allopatric processes, respectively. We constructed the climatic niche of each species using occurrence records and estimated the distance, similarity, and unique fraction of climatic niche between sister species. Calypte species showed moderate niche divergence (Euclidean distance between centroids = 1.94, Sørensen index of similarity = 0.080, unique fraction of hypervolume in C. costae = 0.57, and C. anna = 0.95). However, contrary to expectations, Basilinna species, which diverged under a vicariant scenario, displayed clear niche divergence (Euclidean distance between centroids = 3.78, Sørensen index of similarity = 0.0001, and unique fraction of hypervolume in B. xantusii = 0.98, and B. leucotis = 0.99). We explained dissimilarity in climatic niches between Basilinna species by the ecological divergence between habitats of disjunct populations, which would have also been associated with increased aridity during the Pliocene.

Variations in touch representation in the hummingbird and zebra finch forebrain

Somatosensation is essential for animals to perceive the external world through touch, allowing them to detect physical contact, temperature, pain, and body position. Studies on rodent vibrissae have highlighted the organization and processing in mammalian somatosensory pathways.1,2 Comparative research across vertebrates is vital for understanding evolutionary influences and ecological specialization on somatosensory systems. Birds, with their diverse morphologies, sensory abilities, and behaviors, serve as ideal models for investigating the evolution of somatosensation. Prior studies have uncovered tactile-responsive areas within the avian telencephalon, particularly in pigeons,3,4,5,6 parrots,7 and finches,8 but variations in somatosensory maps and responses across avian species are not fully understood. This study aims to explore somatotopic organization and neural coding in the telencephalon of Anna's hummingbirds (Calypte anna) and zebra finches (Taeniopygia guttata) by using invivo extracellular electrophysiology to record activity in response to controlled tactile stimuli on various body regions. These findings reveal unique representations of body regions across distinct forebrain somatosensory nuclei, indicating significant differences in the extent of areas dedicated to certain body surfaces, which may correlate with their behavioral importance.

The spatiotemporal richness of hummingbird wing deformations.

Animals exhibit an abundant diversity of forms, and this diversity is even more evident when considering animals that can change shape on demand. The evolution of flexibility contributes to aspects of performance from propulsive efficiency to environmental navigation. It is, however, challenging to quantify and compare body parts that, by their nature, dynamically vary in shape over many time scales. Commonly, body configurations are tracked by labelled markers and quantified parametrically through conventional measures of size and shape (descriptor approach) or non-parametrically through data-driven analyses that broadly capture spatiotemporal deformation patterns (shape variable approach). We developed a weightless marker tracking technique and combined these analytic approaches to study wing morphological flexibility in hoverfeeding Anna's hummingbirds (Calypte anna). Four shape variables explained >95% of typical stroke cycle wing shape variation and were broadly correlated with specific conventional descriptors such as wing twist and area. Moreover, shape variables decomposed wing deformations into pairs of in-plane and out-of-plane components at integer multiples of the stroke frequency. This property allowed us to identify spatiotemporal deformation profiles characteristic of hoverfeeding with experimentally imposed kinematic constraints, including through shape variables explaining <10% of typical shape variation. Hoverfeeding in front of a visual barrier restricted stroke amplitude and elicited increased stroke frequencies together with in-plane and out-of-plane deformations throughout the stroke cycle. Lifting submaximal loads increased stroke amplitudes at similar stroke frequencies together with prominent in-plane deformations during the upstroke and pronation. Our study highlights how spatially and temporally distinct changes in wing shape can contribute to agile fluidic locomotion.

Comparing the Contributions of Cutaneous and Respiratory Evaporative Water Loss to Cooling in Anna’s Hummingbirds (Calypte anna) Under Fasted and Fed Conditions

During high-temperature periods, endothermic animals evaporate water from the body surface to cool down. The two primary routes of evaporative water loss are: (1) evaporating water from the skin; and (2) evaporation from the respiratory surfaces during breathing and panting. In smaller avian species like Anna's hummingbirds ( Calypte anna), the higher cutaneous surface area to volume ratio presents an opportunity for the skin to become an important avenue for evaporative cooling, possibly becoming a major site. Additionally, hummingbirds feed on water-rich nectar. As a result, they eliminate surplus water through urination, but significant evaporation can also occur through their skin and respiratory system. Thus, total evaporative water loss could rise significantly for these animals when in a hydrated state. Here, this study investigates the relative importance of the two evaporative water loss routes (cutaneous and respiratory) of Anna's hummingbirds in fed and fasted states. We hypothesize that cutaneous evaporation has a high contribution to total evaporation. Moreover, we predict that evaporative water loss rates are higher under the fed (hydrated) condition relative to fasted states. Metabolic and evaporation rates from the head and the body were measured separately in a partitioned chamber at 30°C. Birds were fed immediately prior to entry into respirometry, and the transition from fed to fasted states was recorded over a two-hour period. Total evaporative water loss nearly doubled in the fed versus fasted condition (1.75 ± 0.85 mg H2O/min and 0.97 ± 0.28 mg H2O/min, respectively), with respiratory water loss being significantly higher in the fed compared to the fasted condition (0.87 ± 0.38 mg H2O/min versus 0.42 ± 0.12 mg H2O/min, respectively). On the other hand, the body had a large, but non-significant, difference in evaporative water loss rates between the fed and fasted conditions (0.88 ± 0.55 mg H2O/min and 0.57 ± 0.17 mg H2O/min, respectively). Importantly, cutaneous evaporative water loss made up half of the total evaporation in both the fed and fasted conditions. These results demonstrate that evaporative water loss is modulated by hydration status at both the respiratory and cutaneous surfaces. Furthermore, cutaneous evaporation makes up approximately the same proportion of total evaporation to other birds at thermoneutral temperatures. Future work manipulating ambient temperature in conjunction with hydration status will further our understanding of the relative importance of cutaneous evaporation for these small-bodied birds. Funded by the Genentech Foundations Scholarship, CSUPERB New Investigator Grant, and San Francisco State University. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Demographic mechanisms and anthropogenic drivers of contrasting population dynamics of hummingbirds

Conserving species requires knowledge of demographic rates (survival, recruitment) that govern population dynamics to allow allocation of limited resources to the most vulnerable stages of target species' life-cycles. Additionally, quantifying drivers of demographic change facilitates enactment of specific remediation strategies. However, knowledge gaps persist in how similar environmental changes lead to contrasting population dynamics through demographic rates. For sympatric hummingbird species, the population of urban-associated partial-migrant Anna's hummingbird (Calypte anna) has increased, yet the populations of Neotropical migrants including rufous, calliope, and black-chinned hummingbirds have decreased. Here, we developed an integrated population model to jointly analyze 25 years of mark-recapture data and population survey data for these four species. We examined the contributions of demographic rates on population growth and evaluated the effects of anthropogenic stressors including human population density and crop cover on demographic change in relation to species' life histories. While recruitment appeared to drive the population increase of urban-associated Anna's hummingbirds, decreases in juvenile survival contributed most strongly to population declines of Neotropical migrants and highlight a potentially vulnerable phase in their life-history. Moreover, rufous hummingbird adult and juvenile survival rates were negatively impacted by human population density. Mitigating threats associated with intensively modified anthropogenic environments is a promising avenue for slowing further hummingbird population loss. Overall, our model grants critical insight into how anthropogenic modification of habitat affects the population dynamics of species of conservation concern.

Sideways maneuvers enable narrow aperture negotiation by free-flying hummingbirds.

Many birds routinely fly fast through dense vegetation characterized by variably sized structures and voids. Successfully negotiating these cluttered environments requires maneuvering through narrow constrictions between obstacles. We show that Anna's hummingbirds (Calypte anna) can negotiate apertures less than one wingspan in diameter using a novel sideways maneuver that incorporates continuous, bilaterally asymmetric wing motions. Crucially, this maneuver allows hummingbirds to continue flapping as they negotiate the constriction. Even smaller openings are negotiated via a faster ballistic trajectory characterized by tucked and thus non-flapping wings, which reduces force production and increases descent rate relative to the asymmetric technique. Hummingbirds progressively shift to the swept method as they perform hundreds of consecutive transits, suggesting increased locomotor performance with task familiarity. Initial use of the slower asymmetric transit technique may allow birds to better assess upcoming obstacles and voids, thereby reducing the likelihood of subsequent collisions. Repeated disruptions of normal wing kinematics as birds negotiate tight apertures may determine the limits of flight performance in structurally complex environments. These strategies for aperture transit and associated flight trajectories can inform designs and algorithms for small aerial vehicles flying within cluttered environments.

Hummingbird ingestion of low-concentration ethanol within artificial nectar.

Both frugivores and nectarivores are potentially exposed to dietary ethanol produced by fermentative yeasts which metabolize sugars. Some nectarivorous mammals exhibit a preference for low-concentration ethanol solutions compared to controls of comparable caloric content, but behavioural responses to ethanol by nectar-feeding birds are unknown. We investigated dietary preference by Anna's Hummingbirds (Calypte anna) for ethanol-enhanced sucrose solutions. Via repeated binary-choice experiments, three adult male hummingbirds were exposed to sucrose solutions containing 0%, 1% or 2% ethanol; rates of volitional nectar consumption were measured over a 3 h interval. Hummingbirds did not discriminate between 0% and 1% ethanol solutions, but exhibited significantly reduced rates of consumption of a 2% ethanol solution. Opportunistic measurements of ethanol concentrations within hummingbird feeders registered values peaking at about 0.05%. Ethanol at low concentrations (i.e. up to 1%) is not aversive to Anna's Hummingbirds and may be characteristic of both natural and anthropogenic nectars upon which they feed. Given high daily amounts of nectar consumption by hummingbirds, chronic physiological exposure to ethanol can thus be substantial, although naturally occurring concentrations within floral nectar are unknown.

Widespread gene flow following range expansion in Anna's Hummingbird.

Anthropogenic changes have altered the historical distributions of many North American taxa. As environments shift, ecological and evolutionary processes can combine in complex ways to either stimulate or inhibit range expansion. Here, we examined the role of evolution in a rapid range expansion whose ecological context has been well-documented, Anna's Hummingbird (Calypte anna). Previous studies have suggested that the C. anna range expansion is the result of an ecological release facilitated by human-mediated environmental changes, where access to new food sources have allowed further filling of the abiotic niche. We examined the role of gene flow and adaptation during range expansion from their native California breeding range, north into Canada and east into New Mexico and Texas, USA. Using low coverage whole genome sequencing we found high genetic diversity, low divergence, and little evidence of selection on the northern and eastern expansion fronts. Additionally, there are no clear barriers to gene flow across the native and expanded range. The lack of selective signals between core and expanded ranges could reflect (i) an absence of novel selection pressure in the expanded range (supporting the ecological release hypothesis), (ii) swamping of adaptive variation due to high gene flow, or (iii) limitations of genome scans for detecting small shifts in allele frequencies across many loci. Nevertheless, our results provide an example where strong selection is not apparent during a rapid, contemporary range shift.

The gyrfalcon (Falco rusticolus) genome.

High-quality genome assemblies are characterized by high-sequence contiguity, completeness, and a low error rate, thus providing the basis for a wide array of studies focusing on natural species ecology, conservation, evolution, and population genomics. To provide this valuable resource for conservation projects and comparative genomics studies on gyrfalcon (Falco rusticolus), we sequenced and assembled the genome of this species using third-generation sequencing strategies and optical maps. Here, we describe a highly contiguous and complete genome assembly comprising 20 scaffolds and 13 contigs with a total size of 1.193 Gbp, including 8,064 complete Benchmarking Universal Single-Copy Orthologs (BUSCOs) of the total 8,338 BUSCO groups present in the library aves_odb10. Of these BUSCO genes, 96.7% were complete, 96.1% were present as a single copy, and 0.6% were duplicated. Furthermore, 0.8% of BUSCO genes were fragmented and 2.5% (210) were missing. A de novo search for transposable elements (TEs) identified 5,716 TEs that masked 7.61% of the F. rusticolus genome assembly when combined with publicly available TE collections. Long interspersed nuclear elements, in particular, the element Chicken-repeat 1 (CR1), were the most abundant TEs in the F. rusticolus genome. A de novo first-pass gene annotation was performed using 293,349 PacBio Iso-Seq transcripts and 496,195 transcripts derived from the assembly of 42,429,525 Illumina PE RNA-seq reads. In all, 19,602 putative genes, of which 59.31% were functionally characterized and associated with Gene Ontology terms, were annotated. A comparison of the gyrfalcon genome assembly with the publicly available assemblies of the domestic chicken (Gallus gallus), zebra finch (Taeniopygia guttata), and hummingbird (Calypte anna) revealed several genome rearrangements. In particular, nine putative chromosome fusions were identified in the gyrfalcon genome assembly compared with those in the G. gallus genome assembly. This genome assembly, its annotation for TEs and genes, and the comparative analyses presented, complement and strength the base of high-quality genome assemblies and associated resources available for comparative studies focusing on the evolution, ecology, and conservation of Aves.

CNTnC and fYTnC2, Genetically Encoded Green Calcium Indicators Based on Troponin C from Fast Animals

NTnC-like green fluorescent genetically encoded calcium indicators (GECIs) with two calcium ion binding sites were constructed using the insertion of truncated troponin C (TnC) from Opsanus tau into green fluorescent proteins (GFPs). These GECIs are small proteins containing the N- and C-termini of GFP; they exert a limited effect on the cellular free calcium ion concentration; and in contrast to calmodulin-based calcium indicators they lack undesired interactions with intracellular proteins in neurons. The available TnC-based NTnC or YTnC GECIs had either an inverted response and high brightness but a limited dynamic range or a positive response and fast kinetics in neurons but lower brightness and an enhanced but still limited dF/F dynamic range. Here, we solved the crystal structure of NTnC at 2.5 Å resolution. Based on this structure, we developed positive NTnC2 and inverted iNTnC2 GECIs with a large dF/F dynamic range in vitro but very slow rise and decay kinetics in neurons. To overcome their slow responsiveness, we swapped TnC from O. tau in NTnC2 with truncated troponin C proteins from the muscles of fast animals, namely, the falcon, hummingbird, cheetah, bat, rattlesnake, and ant, and then optimized the resulting constructs using directed molecular evolution. Characterization of the engineered variants using purified proteins, mammalian cells, and neuronal cultures revealed cNTnC GECI with truncated TnC from Calypte anna (hummingbird) to have the largest dF/F fluorescence response and fast dissociation kinetics in neuronal cultures. In addition, based on the insertion of truncated TnCs from fast animals into YTnC2, we developed fYTnC2 GECI with TnC from Falco peregrinus (falcon). The purified proteins cNTnC and fYTnC2 had 8- and 6-fold higher molecular brightness and 7- and 6-fold larger dF/F responses to the increase in Ca2+ ion concentration than YTnC, respectively. cNTnC GECI was also 4-fold more photostable than YTnC and fYTnC2 GECIs. Finally, we assessed the developed GECIs in primary mouse neuronal cultures stimulated with an external electric field; in these conditions, cNTnC had a 2.4-fold higher dF/F fluorescence response than YTnC and fYTnC2 and was the same or slightly slower (1.4-fold) than fYTnC2 and YTnC in the rise and decay half-times, respectively.

Anna's hummingbird (Calypte anna) physiological response to novel thermal and hypoxic conditions at high elevations.

Many species have not tracked their thermal niches upslope as predicted by climate change, potentially because higher elevations are associated with abiotic challenges beyond temperature. To better predict whether organisms can continue to move upslope with rising temperatures, we need to understand their physiological performance when subjected to novel high-elevation conditions. Here, we captured Anna's hummingbirds - a species expanding their elevational distribution in concordance with rising temperatures - from across their current elevational distribution and tested their physiological response to novel abiotic conditions. First, at a central aviary within their current elevational range, we measured hovering metabolic rate to assess their response to oxygen conditions and torpor use to assess their response to thermal conditions. Second, we transported the hummingbirds to a location 1200 m above their current elevational range limit to test for an acute response to novel oxygen and thermal conditions. Hummingbirds exhibited lower hovering metabolic rates above their current elevational range limit, suggesting lower oxygen availability may reduce performance after an acute exposure. Alternatively, hummingbirds showed a facultative response to thermal conditions by using torpor more frequently and for longer. Finally, post-experimental dissection found that hummingbirds originating from higher elevations within their range had larger hearts, a potential plastic response to hypoxic environments. Overall, our results suggest lower oxygen availability and low air pressure may be difficult challenges to overcome for hummingbirds shifting upslope as a consequence of rising temperatures, especially if there is little to no long-term acclimatization. Future studies should investigate how chronic exposure and acclimatization to novel conditions, as opposed to acute experiments, may result in alternative outcomes that help organisms better respond to abiotic challenges associated with climate-induced range shifts.

DNA metabarcoding reveals broadly overlapping diets in three sympatric North American hummingbirds

ABSTRACT Hummingbirds, a highly diverse avian family, are specialized vertebrate pollinators that feed upon carbohydrate-rich nectar to fuel their fast metabolism while consuming invertebrates to obtain protein. Previous work has found that morphologically diverse hummingbird communities exhibit higher diet specialization on floral resources than morphologically similar hummingbird communities. Due to the difficulties of studying avian diets, we have little understanding whether hummingbirds show similar patterns with their invertebrate prey. Here, we use DNA metabarcoding to analyze floral and invertebrate diets of 3 species of sympatric North American hummingbirds. We collected fecal samples from 89 Anna’s (Calypte anna), 39 Black-chinned (Archilochus alexandri), and 29 Calliope (Selasphorus calliope) hummingbirds in urban and rural localities as well as across an elevational gradient from sea level to 2,500 meters above sea level in California, USA. We found hummingbirds showed high dietary overlap in both invertebrate and plant resources, with few invertebrate and plant families common to most individuals and many families found in only a few individuals. Chironomidae was the most common invertebrate family across all species, and Rosaceae and Orobanchaceae were the most common plant families. Anna’s Hummingbirds had significantly higher invertebrate diet diversity than Black-chinned Hummingbirds when found at the same sites, but we found no difference in plant diet diversity among any of the 3 species. Hummingbirds in urban sites had higher plant diet diversity than in rural sites, but we found no effect of elevation on dietary richness. Our study shows how DNA metabarcoding can be used to non-invasively investigate previously unknown life-histories of well-studied birds, lending insight to community structure, function, and evolution.

A second decade of overwintering hummingbirds in Florida and Alabama

For a second decade, from November 2008 to March 2018, we examined the species diversity, return rate, longevity, and migration routes of hummingbirds overwintering in southern Alabama and the panhandle, northern peninsula, and central peninsula of Florida. We captured and banded 11 species of hummingbirds (n= 1,870 individuals), including 931 Rufous Hummingbirds (Selasphorus rufus; 49%), 583 Ruby-throated Hummingbirds (Archilochus colubris; 31%), 219 Black-chinned Hummingbirds (A. alexandri; 12%), 49 Buff-bellied Hummingbirds (Amazilia yucatanensis), 49 Calliope Hummingbirds (S. calliope), 19 Allen's Hummingbirds (S. sasin), 11 Broad-tailed Hummingbirds (S. platycercus), 6 Broad-billed Hummingbirds (Cynanthus latriostris), 1 Anna's Hummingbird (Calypte anna), 1 Costa's Hummingbird (Calypte costae), and 1 White-eared Hummingbird (Hylocharis leucotis). Site fidelity was high, with 299 hummingbirds of 5 species returning at least once. Two Rufous Hummingbirds migrated to our study area in the autumn from the West (Oklahoma and Texas). One Ruby-throated Hummingbird banded in Lakeland, Florida, was recovered in Utopia, New Brunswick, Canada, suggesting the overwintering Ruby-throated Hummingbirds in our study area are migratory and not sedentary.

Retrospective study on admission trends of Californian hummingbirds found in urban habitats (1991-2016).

BackgroundHummingbirds are frequently presented to California wildlife rehabilitation centers for medical care, accounting for approximately 5% of overall admissions. Age, sex, and reason for admission could impact hummingbird survivability, therefore identification of these factors could help maximize rehabilitation efforts.MethodsMixed-effects logistic regression models were used to identify specific threats to the survival of 6908 hummingbirds (1645 nestlings and 5263 non-nestlings) consisting of five species (Calypte anna, Calypte costa, Selasphorus rufus, Selasphorus sasin, Archilochus alexandri), found in urban settings, and admitted to California wildlife rehabilitation centers over 26 years.ResultsIn total, 36% of birds survived and were transferred to flight cage facilities for further rehabilitation and/or release. Nestlings were more likely to be transferred and/or released compared to adult hummingbirds. After accounting for age, birds rescued in spring and summer were twice as likely to be released compared to birds rescued in the fall. A high number of nestlings were presented to the rehabilitation centers during spring, which coincides with the nesting season for hummingbirds in California, with the lowest number of nestlings presented in fall. Reasons for presentation to rehabilitation centers included several anthropogenic factors such as window collisions (9.6%) and interactions with domesticated animals (12.9%). Survival odds were lower if a hummingbird was rescued in a “torpor-like state” and were higher if rescued for “nest-related” reasons. Evaluation of treatment regimens administered at wildlife rehabilitation centers identified supportive care, including providing commercial nutrient-rich nectar plus solution, to significantly increase hummingbird survivability.DiscussionOur results provide evidence of threats to hummingbirds in urban habitats, based on reasons for rescue and presentation to rehabilitation centers. Reasons for hummingbird admissions to three California wildlife rehabilitation centers were anthropogenic in nature (i.e., being associated with domestic animals, window collisions, and found inside a man-made structure) and constituted 25% of total admissions. There was a clear indication that supportive care, such as feeding a commercial nectar solution, and medical treatment significantly increased the odds of survival for rescued hummingbirds.

IsoDetect: Detection of Splice Isoforms from Third Generation Long Reads Based on Short Feature Sequences

Background: Transcriptome annotation is the basis for understanding gene structures and analysing gene expression. The transcriptome annotation of many organisms such as humans is far from incomplete, due partly to the challenge in the identification of isoforms that are produced from the same gene through alternative splicing. Third generation sequencing (TGS) reads provide unprecedented opportunity for detecting isoforms due to their long length that exceeds the length of most isoforms. One limitation of current TGS reads-based isoform detection methods is that they are exclusively based on sequence reads, without incorporating the sequence information of annotated isoforms. Objective: We aim to develop a method to detect isoforms by incorporating annotated isoforms. Methods: Based on annotated isoforms, we propose a splice isoform detection method called IsoDetect. First, the sequence at exon-exon junctions is extracted from annotated isoforms as “short feature sequences”, which is used to distinguish splice isoforms. Second, we align these feature sequences to long reads and partition long reads into groups that contain the same set of feature sequences, thereby avoiding the pair-wise comparison among the large number of long reads. Third, clustering and consensus generation are carried out based on sequence similarity. For the long reads that do not contain any short feature sequence, clustering analysis based on sequence similarity is performed to identify isoforms. Therefore, our method can detect not only known but also novel isoforms. Result: Tested on two datasets from Calypte anna and Zebra Finch, IsoDetect shows higher speed and good accuracies compared with four existing methods. Conclusion: IsoDetect may become a promising method for isoform detection.

Monitoring through Community Science: Anna’s Hummingbird Winter Range Expansion into Idaho

Increased urbanization and supplementary feeding are implicated in driving the expansion of the range of the Anna’s Hummingbird (Calypte anna). In many areas this range expansion has been well described, but the recent expansion of the northeastern limit of the nonbreeding distribution, in winter in Idaho, has not yet been summarized. Using data from the Idaho Bird Records Committee database and www.eBird.org from 1976 through 2020, we collated records for Idaho and supplemented them with data from a community-science program of monitoring by homeowners. Our additional effort to solicit records from the community shows that database records and feeder observations alone underestimate the number of individuals present in the state. Through banding and color-marking of 58 individual hummingbirds at private residences, we documented six instances of Anna’s Hummingbirds returning to a site in successive winters, found a roughly even sex ratio, and found a ratio of adults to juveniles of about 3:1. Anna’s Hummingbird may now be a sparse year-round resident in parts of Idaho.

Macroecological patterns of resource use in resident and migratory hummingbirds

Hummingbirds (Family Trochilidae) are key pollinators in several biodiversity hotspots, including the California Floristic Province in North America. Relatively little is known about how hummingbird diets change throughout the year, especially with regard to how migratory hummingbirds affect resident hummingbirds at stopover sites. In this study, we examine how hummingbird species, migratory status, sex, geographic region and local plant diversity influence floral resource use before, during, and after an influx of migratory hummingbirds (primarily Rufous hummingbirds, Selasphorus rufus) across California. We expected distinct floral resource use based upon species’ migratory status (resident vs. migrant), sex, sampling period, and geographic region. We employed DNA metabarcoding to detect plant DNA in hummingbird fecal samples to analyze diet diversity, composition, overlap, and interaction networks. We found significant effects of sex, sampling period, and migratory status on the alpha and beta diversity of plant taxa present in fecal samples. Analyses of Anna's hummingbirds (Calypte anna) alone revealed that female fecal samples contained higher plant species richness. In addition to hummingbird-pollinated plants, fecal samples also contained non-ornithophilous plants and species of agricultural importance. Diet overlap and plant-pollinator network analyses revealed high overlap in plant taxa used between hummingbird species, and networks were more connected, less nested, and less specialized than null models. DNA metabarcoding is minimally invasive and provides a detailed view of hummingbird diet, permitting large-scale studies. Insights into hummingbird diets are especially valuable given the logistical difficulties of directly observing floral visitation and foraging across broad temporal and spatial scales.

LETHAL INTRASPECIFIC BEHAVIOR BY ANNA'S HUMMINGBIRD

Intraspecific agnostic behavior is well-known in hummingbirds; however, lethality in North American species is unreported. Here we describe an observation of a lethal attack by a male Anna's Hummingbird (Calypte anna) on another male of the same species during mid-winter near the northern extent of the species' winter range.

Concentrations of Retinol and α-Tocopherol in Tissue Samples From Anna's Hummingbirds (Calypte anna).

Retinol (vitamin A) and α-tocopherol (vitamin E) concentrations were measured in tissue samples (liver, heart, pectoral muscle, and brain) from Anna's Hummingbirds (Calypte anna). Hummingbirds were after-hatch year birds that were sourced from various rehabilitation centers throughout California. Tissues samples were analyzed using high-performance liquid chromatography (HPLC). Minimum, maximum, mean, standard deviation (SD), and median ppm concentrations were calculated for each vitamin and tissue sample type. A novel analytical method was developed to analyze small mass tissue samples, with the smallest sample mass being 0.05 g for which analysis can be performed. Mean ± standard deviation (SD) concentrations of retinol in hummingbird livers, hearts, and pectoral muscle samples were 269.0 ± 216.9 ppm, 1.8 ± 2.2 ppm, and 0.3 ± 0.1 ppm, respectively. Mean ± SD α-tocopherol concentrations were 6.9 ± 4.6 ppm, 5.5 ± 4.0 ppm, 3.7 ± 2.2 ppm, and 9.1 ± 3.2 ppm for liver, heart, pectoral muscle, and brain samples, respectively. Vitamin concentrations from varying tissue types were compared to determine which were best associated with liver concentrations, the most commonly analyzed tissue for these vitamins. For both retinol and α-tocopherol, heart samples were most strongly associated with the liver samples. The results of this study provide baseline retinol and α-tocopherol concentrations in different tissue types from Anna's hummingbirds. These baseline values may be utilized in conservation efforts to avoid hypervitaminosis and hypovitaminosis of rehabilitated and/or captive hummingbirds by providing guidelines for nutritional targets which could be assessed on post-mortem examinations. Post-mortem examination of birds and measurement of vitamin concentrations in tissues may allow for dietary changes that aid captive hummingbirds.

Ontogeny of vocal learning in a hummingbird

Hummingbirds have evolved to learn song from conspecifics independently of other bird clades that show this behaviour, such as parrots and oscine passerines. Little is known about how vocal learning occurs in hummingbirds. To determine the conditions necessary and sufficient for learning, we raised young Costa's hummingbirds, Calypte costae, in isolation in two experiments. The first experiment tested whether three pairs of males, raised either in silence or with only acoustic exposure to song, would produce wild-type song. They did not: each pair instead produced ‘isolate’ songs dissimilar to wild-type song. Each pair produced a unique isolate song with subtle differences from other pairs. Thus, exposure to song alone is insufficient to trigger learning. The second experiment tested whether individually housed males would learn songs when exposed to a live male model plus playback of song (1 h/day, 3 days/week, for 3 months). Seven of eight birds that heard Costa's-like songs learned to sing their tutor song. Each individual produced song that matched idiosyncratic features of the individual songs played to them, demonstrating attention to, and learning of the exact acoustic stimuli received. Two control birds exposed to acoustically dissimilar Anna's hummingbird, Calypte anna, song developed isolate song. Thus, exposure to species-specific song with a conspecific male is sufficient to trigger learning. The sensory phase began after fledging, around 23 days posthatch, and the sensorimotor phase began around 50 days posthatch, when singing also began. Song crystallization occurred approximately 125 days posthatch. Costa's also appear to be statistical learners; they learn to sing the songs they hear most often. Hummingbird song ontogeny appears to be similar to song ontogeny in passerines, despite its convergent origin.