Avian Egg Research Articles

Primordial germ cells isolated from individual embryos of red junglefowl and indigenous pheasants of Thailand

Interspecific germline chimerism mediated by transplantation of primordial germ cells (PGCs) of wild species to domestic hosts promises the conservation of wild birds. Cryopreservation of avian eggs and embryos is impracticable, and currently only frozen PGCs enable conservation of both the male and female descendants. Purebred offspring have been obtained from germline chimeras of wild avian species, proving the feasibility of such technology. In vitro propagation has been optimized for avian PGCs of domestic species; however, evidence is rather limited for successful isolation as well as long-term culture from a single embryo of wild species. With accelerating biodiversity loss, we have committed to preserving current genetic resources by freezing PGCs isolated from individual embryos in addition to their genetic material. We have devised a reliable protocol for the isolation and proliferation of PGCs from wild fowls in the family Phasianidae that are conserved in captive breeding (red junglefowl, bar-tailed pheasant, kalij pheasant, Siamese fireback pheasant, and silver pheasant). We obtained individual isolates of cultured circulating PGCs (49.7%, 79/155) as well as tissue PGCs (92.9%, 144/155). The specific co-culture conditions of autologous embryonic cells, without additional growth factors, facilitated the proliferation of so-called tissue PGCs (the remaining PGCs in embryonic tissue following blood aspiration). Only circulating PGCs left in blood vessels and of PGCs migrating to developing gonads have been previously reported. However, the present study is the first to report on the harvest of ectopic PGCs. The defined conditions sustained continuous proliferation of tissue PGCs for at least six months and maintained PGC identity following cryopreservation. Cultured tissue PGCs of these wild species were extensively characterized for their expression of the germ cell-specific proteins, chicken vasa homolog (CVH) and deleted in azoospermia-like (DAZL), as well as the ability to colonize chicken embryonic gonads. The novel protocol is practical for generating enough PGCs for cryopreservation, transplantation, and additionally, it enables isolation of PGCs from both blood circulation and embryonic tissue simultaneously. For conservation purposes, this approach is potentially applicable more widely to other non-domestic birds than those in the family Phasianidae that were investigated in the present study.

The Trichohyalin-Like Protein Scaffoldin Is Expressed in the Multilayered Periderm during Development of Avian Beak and Egg Tooth.

Scaffoldin, an S100 fused-type protein (SFTP) with high amino acid sequence similarity to the mammalian hair follicle protein trichohyalin, has been identified in reptiles and birds, but its functions are not yet fully understood. Here, we investigated the expression pattern of scaffoldin and cornulin, a related SFTP, in the developing beaks of birds. We determined the mRNA levels of both SFTPs by reverse transcription polymerase chain reaction (RT-PCR) in the beak and other ectodermal tissues of chicken (Gallus gallus) and quail (Coturnix japonica) embryos. Immunohistochemical staining was performed to localize scaffoldin in tissues. Scaffoldin and cornulin were expressed in the beak and, at lower levels, in other embryonic tissues of both chickens and quails. Immunohistochemistry revealed scaffoldin in the peridermal compartment of the egg tooth, a transitory cornified protuberance (caruncle) on the upper beak which breaks the eggshell during hatching. Furthermore, scaffoldin marked a multilayered peridermal structure on the lower beak. The results of this study suggest that scaffoldin plays an evolutionarily conserved role in the development of the avian beak with a particular function in the morphogenesis of the egg tooth.

Exposure to a mimetic or non-mimetic model avian brood parasite egg does not produce differential glucocorticoid responses in an egg-accepter host species

Avian obligate brood parasitism, a reproductive strategy where a parasite lays its egg into the nest of another species, imposes significant fitness costs upon host parents and their offspring. To combat brood parasitism, many host species recognize and reject foreign eggs (rejecters), but others are accepters that raise the parasitic progeny. Some accepter hosts may be unable to grasp or pierce parasitic eggs even if they recognize them as foreign eggs in the clutch, whereas other accepters may not have evolved the cognitive skillsets to recognize dissimilar eggs in the nest. Here we assessed the endocrine responses of an accepter host species to model parasitic eggs to address these two alternatives. We experimentally parasitized nests of a locally common host of the brood-parasitic brown-headed cowbird (Molothrus ater), the prothonotary warbler (Protonotaria citrea; a cowbird-egg accepter), with a mimetic or non-mimetic model cowbird-sized egg. Our goal was to determine whether they perceived the non-mimetic egg as a greater stressor by measuring circulating corticosterone levels. We added eggs to nests during the incubation stage and obtained blood plasma samples from females on the nest 2 h later, using females with unmanipulated clutches as controls. Incubating females showed no differences in baseline plasma corticosterone levels between our different treatments. We conclude that exposure to foreign eggs does not activate the hypothalamic–pituitary–adrenal axis of prothonotary warbler hosts in this experimental paradigm.

Mechanical Separation and Protein Solubilization of the Outer and Inner Perivitelline Sublayers from Hen's Eggs

The perivitelline layer that surrounds the egg yolk plays a fundamental role in fertilization, in egg defense, and in the development of the avian embryo. It is formed by two proteinaceous sublayers that are tightly associated and formed by distinct female reproductive organs. Both structures are assumed to have their own functional specificities, which remain to be defined. To characterize the function of proteins composing each sublayer, the first challenge is to establish the conditions that would allow for the mechanical separation of these two intricate layers, while limiting any structural damage. The second step is to optimize the experimental conditions to facilitate protein solubilization from these two sublayers, for subsequent biochemical analyses. The efficiency of this approach is assessed by analyzing the protein profile of each sublayer by Sodium Dodecyl Sulfate-Poly-Acrylamide Gel Electrophoresis (SDS-PAGE), which is expected to be distinct between the two structures. This two-step procedure remains simple; it requires classical biochemical equipment and reagents; and is compatible with further in-depth proteomics. It may also be transposed to other avian eggs for comparative biology, knowing that the structure and the composition of the perivitelline layer has been shown to have species-specific features. In addition, the non-denaturing conditions developed for sublayers separation (step 1) allow their structural analyses by scanning and transmission electron microscopy. It may also constitute the initial step for subsequent protein purification to analyze their respective biological activities and 3D structure, or to perform further immunohistochemical or functional analyses. Such studies would help to decipher the physiological function of these two sublayers, whose structural and functional integrities are determinant criteria of the reproductive success.

Mechanical Separation and Protein Solubilization of the Outer and Inner Perivitelline Sublayers from Hen's Eggs.

The perivitelline layer that surrounds the egg yolk plays a fundamental role in fertilization, in egg defense, and in the development of the avian embryo. It is formed by two proteinaceous sublayers that are tightly associated and formed by distinct female reproductive organs. Both structures are assumed to have their own functional specificities, which remain to be defined. To characterize the function of proteins composing each sublayer, the first challenge is to establish the conditions that would allow for the mechanical separation of these two intricate layers, while limiting any structural damage. The second step is to optimize the experimental conditions to facilitate protein solubilization from these two sublayers, for subsequent biochemical analyses. The efficiency of this approach is assessed by analyzing the protein profile of each sublayer by Sodium Dodecyl Sulfate-Poly-Acrylamide Gel Electrophoresis (SDS-PAGE), which is expected to be distinct between the two structures. This two-step procedure remains simple; it requires classical biochemical equipment and reagents; and is compatible with further in-depth proteomics. It may also be transposed to other avian eggs for comparative biology, knowing that the structure and the composition of the perivitelline layer has been shown to have species-specific features. In addition, the non-denaturing conditions developed for sublayers separation (step 1) allow their structural analyses by scanning and transmission electron microscopy. It may also constitute the initial step for subsequent protein purification to analyze their respective biological activities and 3D structure, or to perform further immunohistochemical or functional analyses. Such studies would help to decipher the physiological function of these two sublayers, whose structural and functional integrities are determinant criteria of the reproductive success.

Abnormal eggs of the Common Guillemot Uria aalge: the role of stress

This study documents several types of abnormal eggs of the Common Guillemot Uria aalge collected from Bempton Cliffs, Yorkshire, UK, during the 1900s and currently housed in the Natural History Museum at Tring, Hertfordshire, UK. These eggs are abnormal in terms of size, shape, colour and surface topography. We suggest that the incidence of these abnormal eggs was relatively high because the manner in which they were collected — repeated visits to the breeding colony — caused considerable disturbance and stress that in turn disrupted the normal process of egg formation. We discuss the possibility of using abnormalities in avian eggs as a measurement of environmental stress.

Evaluation of host systems for efficient isolation and propagation of duck Tembusu virus

ABSTRACT Several phylogenetic clusters of duck Tembusu virus (DTMUV) that caused outbreaks in ducks in Asia have been identified since its emergence in 2010, highlighting the need for an efficient host system that can support isolation of all circulating phylogenetic clusters of DTMUV. In this study, various host systems, including different avian embryonated eggs (duck and chicken) and cell cultures (primary duck embryo fibroblast (DEF), primary chicken embryo fibroblast (CEF), baby hamster kidney (BHK-21), African green monkey kidney (Vero) and Aedes albopictus clone C6/36 (C6/36) cells), were evaluated and compared for their ability to support DTMUV isolation and propagation. Our results showed that all host systems were susceptible to DTMUV infection; however, BHK-21 and primary DEF cells supported more efficient replication of DTMUV compared to the other host systems. BHK-21 cells had the highest DTMUV isolation rate when tested with experimental and field clinical samples. All circulating phylogenetic clusters of DTMUV, including clusters 1, 2 and 3, were successfully isolated from duck clinical samples using BHK-21 cells. In conclusion, our findings supported the use of BHK-21 cells as a host system for primary isolation of all circulating phylogenetic clusters of DTMUV from duck clinical samples. This study highlights the importance of selecting the most appropriate host system for efficient isolation and propagation of DTMUV from duck clinical samples. RESEARCH HIGHLIGHTS DTMUV replicated more efficiently in BHK-21 and primary DEF cells than in other host systems tested. BHK-21 cells had the highest DTMUV isolation rate. All DTMUV phylogenetic clusters were successfully isolated from the samples using BHK-21 cells. BHK-21 cells were the most efficient host system for DTMUV isolation.

Evaluación de un protocolo automatizado para la obtención de medidas morfométricas de huevos de aves a partir de fotografías digitales

Evaluation of a protocol for automated extraction of morphometric measurements from avian eggs using digital photography As many ecomorphological studies are limited by the time required to gather manual measurement data, automatizing the process is an important focus of methodological innovations. We developed, implemented and validated a protocol for the semi–automated extraction of a set of morphometric variables of egg size and shape from digital pictures. The protocol was implemented in R language as a web app called OvometriK. After binarizing and calibrating images, this protocol uses geometric and trigonometric functions to calculate eleven egg variables. We tested calculations in several ways, assuming contour continuity or using voxel counts. Application was validated with geometric shapes and 30 manually–measured chicken eggs. Mathematical validation with spheres showed that the algorithm provided high precision diameter measures, with a correlation of 99.9 %. Average estimation error was 1.4 %. The mathematical volume estimation was underestimated by 27 %, while voxels were underestimated by only 6 %. Differences between manual egg measurements of diameters and those obtained from images was less than 3 mm (4 %). Correlation between estimated volume and measured by silica gel filling was higher than 90 % using the voxel count method. Neither inclination angle or picture resolution had significant effects on precision (3.2 % maximum difference). Measures showed high repeatability and represent a significant saving in processing time. This new protocol represents an improvement on previous programs regarding limitations of platform, accessibility and number of variables. Furthermore, its flexibility and openness means it can be adapted to other specific applications.

Artificial nest experiment reveals inter-guild facilitation in duck nest predation

Predation is the most important factor limiting Northern Hemisphere duck nest survival. While most avian nest predators take only eggs, mammalian predators also pose mortality risks to incubating ducks. Partial depredation of duck clutches elevates subsequent nest desertion probability, especially where eggshells provide evidence of the depredation event. Avian predators tend to consume egg contents at the nest, which may not intrinsically pose a risk to hens, but leave egg remains. This begs the question: does primary avian depredation of duck eggs actively facilitate nest visits by secondary mammalian predators, with potential survival consequences for the remaining clutch, but also sitting hens? We created 335 artificial duck nests in Finland and Denmark during 2017–2019 and monitored them for seven days using wildlife cameras to test whether initial avian predators breaking the eggs facilitated visits by secondary mammalian predators in various duck breeding habitats. Visitation rates of mammalian predators likely to take duck hens increased significantly after avian egg depredation events that involved egg breakage, although rates differed between habitats. Most common mammalian nest visitors at nests depredated by avian predators were red fox (Vulpes vulpes), followed by two alien species, raccoon dog (Nyctereutes procyonoides) and American mink (Neovison vison). Although we should be prudent about extrapolating conclusions from artificial experiment to real duck nests, our results strongly suggest inter-guild facilitation between avian and mammalian predators at duck nest sites. We suggest that by breaking the eggs, avian predators may offer olfactory, visual or other cues for secondary predators. Nest visits by mammalian predators could potentially increase egg depredation risk, but also elevate mortality risks for incubating females, dependent upon prevailing predator guilds.

Avian egg collections: museum collection bias driven by shape and size

Avian eggs exhibit substantial intra‐ and interspecific variation in shape, size and colour. Considerable efforts have been made to better understand the evolutionary drivers behind such variation, often using museum egg collections. Usually it is assumed that museum collections accurately represent the variation seen in natural populations, but this may not be the case if there is collection bias. Collection bias may lead to the over‐representation of certain egg traits in collections, due to the aesthetic (or other) preferences of collectors. The aim of this study is to begin to look for the occurrence of potential collection bias in museum egg collections by comparing three shape indices (pointedness/asymmetry, elongation and polar asymmetry) and egg volume between subsets of eggs in museum collections with those of recently sampled eggs in the field for three different bird species: common guillemot Uria aalge, razorbill Alca torda and northern fulmar Fulmarus glacialis. We found no evidence of collection bias in our sampled razorbill and northern fulmar museum collection eggs, but some evidence for a bias in sampled museum collection eggs of common guillemots. Since the guillemot's egg differs from most bird eggs in being pyriform, we suggest that collection bias by historic egg collectors may be more prevalent in species with extreme egg traits. Researchers using museum egg collections to examine questions relating to egg shape should be aware of collection bias risks and consider how to minimise the effect of these possible biases on accumulated datasets.

Digital imaging assisted geometry of chicken eggs using Hügelschäffer's model

Geometrical description of the egg shape is of a great importance in a variety of studies and can be instrumental in predicting quality traits of table and hatching poultry eggs. Importantly, developments of non-destructive oomorphological models can drive novel insights in engineering and physical science and lead to new egg-related technologies and egg sorting systems for poultry industry. We attempted to test the Hugelschaffer's egg model according to which an egg profile curve can be transformed from an ellipse using a specific parameter w. For this purpose, two-dimensional digital imaging and follow-up image processing techniques of chicken eggs were employed. The formulae for recalculation of the egg volume and surface area were consequently deduced from the Hugelschaffer's equation. Eventually, we refined the Hugelschaffer's egg model and proved its applicability for defining the contours of hen's eggs. For practical use in poultry industry and food engineering, the proposed non-destructive methodology can be contributory in defining accurately the contour of any avian egg and determining such characteristics of the egg shape as volume, surface area, etc., with an expected potential in designing automated systems in poultry industry and in egg-related applications in biology, physical science, engineering and other areas.

Avian Egg Timers: Female Cowbirds Judge Past, Present, and Future When Making Nest Parasitism Decisions

The cognitive demands associated with brood parasitism are substantial. Not only must female parasites locate nests and assess their suitability for parasitism, they must also time parasitism to correspond with the breeding behavior of the host. Keeping track of the reproductive state of hosts for a variety of nests allows the parasite to select a nest where their egg can be incubated successfully. Thus, nest selection decisions are integral to obligate brood parasites’ reproductive success. In captive breeding flocks of brown-headed cowbirds (Molothrus ater), I studied (1) females’ abilities to time a host nest’s readiness for parasitism and (2) the timing of females’ nest selection decisions. I found that cowbirds can attend to the amount of time that elapsed since a host egg was added to a nest and can use that information to choose a nest for parasitism. Females made their choice of nest during prospecting the day before they laid, and, once decided, they did not update their decisions on the day of laying. Taken together, the results reveal that female cowbirds process substantial amounts of information about location, time, number, and rate in order to plan for future parasitism. This research program reveals that species-typical decisions integrate a variety of general and specialized cognitive abilities to allow females to behave adaptively and maximize reproductive success.

Cryo-EM Structure of Native Human Uromodulin, a Zona Pellucida Module Polymer

Assembly of extracellular filaments and matrices mediating fundamental biological processes such as morphogenesis, hearing, fertilization and antibacterial defense is driven by a ubiquitous polymerization module known as zona pellucida (ZP) “domain”. Despite the conservation of this element from hydra to human, no information is available on the filamentous conformation of any ZP module protein. Here we report the cryo-electron microscopy structure of uromodulin (UMOD)/Tamm-Horsfall protein, the most abundant protein in human urine and an archetypal ZP module-containing molecule, in its mature homopolymeric state. UMOD forms a one-start helix with an unprecedented 180-degree twist between subunits enfolded by interdomain linkers that have completely reorganized as a result of propeptide dissociation. Lateral interaction between filaments in the urine generates sheets exposing a checkerboard of binding sites to capture uropathogenic bacteria, and UMOD-based models of mammalian and avian heteromeric egg coat filaments identify a common sperm-binding region at the interface between subunits.

Yolk immunoreactive corticosterone does not relate with maternal fecal glucocorticoids in a captive population of Greater Rheas (Rhea americana)

Maternal corticosterone in avian egg yolks may signal and prepare offspring for the prevailing conditions. However, this adjustment requires flexible mechanisms of regulation that lead to the absence of a correlation between hormone levels in the mother and the yolk. In this study, we examined whether yolk immunoreactive corticosterone and maternal glucocorticoid levels are related in the Greater Rhea (Rhea americana), a ratite species with a communal nesting system. During the intermediate laying period of the season, we carried out daily nest-observations to collect those eggs in which the laying female was identified. We also collected fresh fecal samples from the identified laying females to quantify their fecal glucocorticoid metabolite (FGM) levels during the yolk formation period (7 days before egg-laying). Yolk immunoreactive corticosterone and FGM levels were measured using a corticosterone125 radioimmunoassay kit, previously validated for the species. No significant relationships were found between female FGM and yolk immunoreactive corticosterone. Although futures studies are needed to elucidate the underlying mechanisms, our results suggest that Greater Rhea female may be able to control the distribution of corticosterone into their yolks, supporting the idea that this hormone may be mediating an adaptive maternal effect.

Effects of sublethal application of Deepwater Horizon oil to bird eggs on embryonic heart and metabolic rate.

Following large crude oil spills, oil from feathers of brooding birds and oiled nesting material can transfer to eggs, resulting in reduced embryonic viability for heavily oiled eggs. Eggs may also be subjected to trace or light oiling, but functional teratogenic effects from sublethal crude oil exposure have not been examined. We assessed whether sublethal application of weathered Deepwater Horizon crude oil to the eggshell surface alters heart rate and metabolic rate in Zebra Finch (Taeniopygia guttata) embryos. We first determined sublethal applications with a dosing experiment. Embryo viability for eggs exposed to 5 μL or more of crude oil decreased significantly. We conducted a second experiment to measure heart rate and metabolic rate (CO2 production) 5 and 9 d after 1 sublethal application of crude oil to eggshells on day 3 of incubation. One application of 1.0 or 2.5 µL of crude oil reduced embryonic heart rate and metabolic rate on day 12 of incubation. Using unfertilized eggs, we measured the transfer of polycyclic aromatic hydrocarbons (PAHs) from the eggshell surface to egg contents 9 d after a single application of sublethal crude oil. Our results suggest avian eggs externally exposed to small amounts of crude oil may exhibit protracted embryonic development and impaired postnatal cardiac performance.

Ca2+ dependent flocculation efficiency of avian egg protein revealed unique surface specific interaction with kaolin particles: A new perception in bioflocculant research

In this study, the flocculation efficiency (FE) of avian egg-white was investigated as a protein rich natural bioflocculant to flocculate kaolin dispersion. The FE was optimized under various conditions. Its cation dependent interaction with kaolin basal planes was also investigated through FT-IR, EEM, Zeta Potential and SEM-EDX techniques. The maximum flocculation rate (91 %) was observed at the CaCl2 dosage of 100 ppm and at pH 6.0 due to the electrostatic interaction of Ca2+ ions with the gibbsite basal planes of kaolin at this condition. Among several monovalent, divalent and trivalent cations, Ca2+ alone showed higher FE than other cations. It showed the unique interaction of Ca2+ ions with Avian Egg-white Flocculant (AVGF) functional groups. EEM study was used to reveal the involvement of such functional groups. The results revealed that the carboxylate groups instead of aromatic amino acid side chains are involving in such interaction. This hypothesis was further confirmed in wastewater flocculation experiments in which the Ca2+ and AVGF complex showed weak interaction with suspended particles of domestic wastewater.

Toxicogenomic Assessment of Complex Chemical Signatures in Double-Crested Cormorant Embryos from Variably Contaminated Great Lakes Sites.

Using omics approaches to monitor complex environmental mixtures is challenging. Previously, we evaluated in vitro transcriptomic effects of complex organic extracts derived from avian eggs. However, there is a lack of studies using wild species that are naturally exposed to contaminant mixtures. Here, we examined polychlorinated biphenyl (PCB) and polybrominated diphenyl ether (PBDE) residues and gene expression in embryonic liver tissue of double-crested cormorants (Phalacrocorax auritus) collected from six variably contaminated colonies. Colonies near industrialized areas were distinguished from less contaminated sites based on their PCB and PBDE concentrations. The most variably expressed genes between sites were involved in pathways including, xenobiotic metabolism (e.g., Cyp1a4), lipid/bile acid homeostasis (e.g., Lbfabp), and oxidative stress (e.g., Mt4). Hierarchical clustering, based on relative gene expression, revealed a grouping pattern similar to chemical residue concentrations. Further, partial least squares regression analysis was used to estimate chemical concentrations from transcriptomics data. PCB 155 and BDE 47 showed the highest slopes (0.77 and 0.69, respectively) fitted by linear regression of measured and estimated chemical concentrations. The application of transcriptomics to a wild avian species, naturally exposed to complex chemical mixtures and other stressors, represents a promising means to distinguish and prioritize variably contaminated sites.

Cases of foreign egg adoption by Brown Skuas Stercorarius antarcticus on Marion and Gough Islands

Ground-nesting birds, such as large gulls and skuas, are known to include a variety of items in their nests as pseudo-eggs, as well as to adopt the eggs of conspecifics or other species. Three hypotheses have been proposed to explain this phenomenon: incubation stimulus, mistaken food, and mistaken egg hypotheses. Of 657 Brown Skua Stercorarius antarcticus nests surveyed on Marion Island during the 2017/18 breeding season, two instances of egg adoption were recorded. Another case of egg adoption was found in a Brown Skua nest on Gough Island in the 2018/19 breeding season. It is likely that all three were cases of mistaken food. This study adds further evidence to the assumption that avian egg predators such as skuas and gulls which are able to steal and carry an egg unbroken in flight, are at risk of this maladaptive behaviour.

Avian egg and nestling detection in the wild: should we rely on visual models or behavioural experiments?

The fields of avian egg and nestling colour detection have rapidly advanced owing to the application of visual models, which have allowed assessing of evolutionary questions considering receiver perception. Here, I first review the literature aiming to identify patterns of avian visual model usage. Second, I elaborate on limitations in the application of the receptor-noise limited perceptual (RNL hereafter) model. A systematic literature review revealed that the RNL model was the most used approach (81% of studies) in the field, and that most studies (76%) were concerned with classic evolutionary questions in avian brood parasitism. Some known limitations of the RNL model deal with model assumptions and parameterization, or, a poor consideration of post-detection neural processes. Others, however, are specific of the fields of egg and nestling discrimination and deal with the highly variable nature of ambient light at the nests, the complex colour design of eggs and nestlings, the multi-dimensional nature of perception, and the possible implication of learning. I, therefore, conclude that visual models should be used with caution to establish inference about egg and nestling discrimination, and rather be used to provide reasonable hypotheses which need to be validated with behavioural experiments. This article is part of the theme issue 'Signal detection theory in recognition systems: from evolving models to experimental tests'.

Estimating egg mass–body mass relationships in birds

Abstract The mass of a bird’s egg is a critical attribute of the species’ life history and represents a fundamental component of reproductive effort. Indeed, the tradeoff between the number of eggs in a clutch and clutch mass lies at the heart of understanding how environmental attributes such as nest predation or adult mortality influence reproductive investment. However, egg masses have not been reported for the majority of avian species. We capitalized on the strong allometric relationship between avian body mass and egg mass to produce egg mass estimates for over 5,500 species previously lacking such information. These estimates are accompanied by measures of the robustness of the regressions used to produce them (e.g., sample size, root mean square error [RMSE] of estimation, coefficient of determination, and degree of extrapolation), thus allowing independent evaluation of the suitability of any estimate to address a particular research question relating to avian life history. Most estimates (~5,000) were based on family-level egg mass–body mass regressions, with the remainder derived from other relationships such as ordinal regressions. We compared estimating regressions based on adult vs. female body masses and, after finding little difference between the 2, based our final estimates on adult masses as those were more numerous in the literature. What small differences between adult- and female-based regressions that did occur were not related to sexual size dimorphism across families. These new estimates, coupled with ~5,000 egg masses reported in the literature, provide a foundation of over 10,000 species for wider investigations assessing variation in reproductive effort in birds over a broad array of ecological and evolutionary contexts.